LOCUS       pLenti-CMV-HA-LZTR1-G248R-puro       10433 bp ds-DNA     circular SYN 24-JAN-2019
DEFINITION  LMBP 11174.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct

COMMENT     VNTDATE|867456000|
COMMENT     VNTDBDATE|867456000|
COMMENT     VNTNAME|pLenti-CMV-HA-LZTR1-G248R-puro|
FEATURES             Location/Qualifiers
     CDS             295..894
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
     LTR             1032..1265
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1337..1458
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      1498..1633
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(1654..1672)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1682..1698)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1840..2295
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2321..2425
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2426..3286
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
     rep_origin      3457..4045
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4333..4354
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4369..4399
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4407..4423
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     4431..4447
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4468..4486
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4514..4740
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4741..4921
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4968..5093
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5586..5819
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    6315..6431
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        6454..6757
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6758..6961
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             7050..7076
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
     CDS             7575..7586
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
     regulatory      7939..7948
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    9606..10194
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(10077..10088)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
     promoter        join(10208..10433,1..274)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
ORIGIN
        1 aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa
       61 gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca
      121 gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg
      181 cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc
      241 gctgttctcc tcttcctcat ctccgggcct ttcgacctgc agcccaagct taccatgacc
      301 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
      361 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
      421 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
      481 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
      541 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
      601 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
      661 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
      721 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
      781 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
      841 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacgcccg
      901 ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc tcgagggccc
      961 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     1021 aaagggggga ctggaagggc tagctcactc ccaacgaaga caagatctgc tttttgcttg
     1081 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     1141 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     1201 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     1261 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     1321 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     1381 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     1441 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc
     1501 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     1561 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     1621 ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct
     1681 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     1741 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     1801 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     1861 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     1921 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     1981 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2041 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2101 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     2161 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     2221 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     2281 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     2341 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     2401 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     2461 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     2521 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     2581 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     2641 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     2701 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     2761 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     2821 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     2881 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     2941 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     3001 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     3061 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     3121 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     3181 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     3241 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     3301 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     3361 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     3421 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3481 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3541 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3601 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3661 atacctcgct ctgctaatcc tgttaccagt ggctgatgcc agtggcgata agtcgtgtct
     3721 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3781 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3841 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3901 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3961 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4021 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4081 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     4141 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     4201 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     4261 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     4321 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     4381 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     4441 ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga
     4501 gctgcaagct taatgtagtc ttatgcaata ctcttgtagt cttgcaacat ggtaacgatg
     4561 agttagcaac atgccttaca aggagagaaa aagcaccgtg catgccgatt ggtggaagta
     4621 aggtggtacg atcgtgcctt attaggaagg caacagacgg gtctgacatg gattggacga
     4681 accactgaat tgccgcattg cagagatatt gtatttaagt gcctagctcg atacataaac
     4741 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     4801 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     4861 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     4921 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
     4981 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
     5041 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
     5101 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
     5161 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
     5221 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
     5281 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
     5341 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
     5401 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
     5461 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
     5521 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
     5581 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
     5641 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
     5701 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     5761 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     5821 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     5881 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     5941 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     6001 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     6061 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     6121 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     6181 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     6241 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatctcga
     6301 cggtatcggt taacttttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga
     6361 atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa
     6421 ttcaaaattt tatcgataag cttgggagtt ccgcgttaca taacttacgg taaatggccc
     6481 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     6541 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     6601 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     6661 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     6721 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
     6781 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     6841 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     6901 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     6961 tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag
     7021 aagacaccga ctctagagcc gccaccatgt acccatacga tgttccagat tacgctatgg
     7081 ctggaccggg cagcacgggg gggcagatcg gggctgcggc cctggcaggc ggcgcgcggt
     7141 ccaaggtagc cccgagcgtg gacttcgacc atagctgctc ggacagtgtc gagtacctga
     7201 cgctcaactt cgggcccttc gaaacagtgc atcgctggcg gcgcctcccg ccctgcgacg
     7261 agtttgtagg tgctaggcgc agcaagcaca cagtggtggc ctataaagat gccatttatg
     7321 tatttggtgg agacaatggg aagaccatgc tcaatgacct cctgcggttc gatgtgaaag
     7381 actgctcctg gtgcagggcc tttaccactg ggaccccacc ggccccccgt taccaccact
     7441 cggccgtcgt ctatgggagc agcatgtttg tctttggggg ttacactggg gacatttatt
     7501 ccaattctaa cttgaagaat aaaaacgacc tctttgaata caagtttgca actggccagt
     7561 ggacggagtg gaaaattgaa ggacggttgc cagtcgctag gtcagcccat ggggccacgg
     7621 tgtacagtga caagctgtgg atctttgctg gctatgacgg caacgccagg ttgaatgaca
     7681 tgtggacaat tggcctccag gaccgagagc tcacctgctg ggaggaggtg gcccagagtg
     7741 gcgagatccc cccatcttgc tgcaacttcc ccgtggctgt gtgccgggac aagatgtttg
     7801 tattctctgg gcaaagcaga gccaaaataa ccaacaacct cttccagttt gaattcaagg
     7861 acaagacgtg gacacgcatc ccaactgaac acctgctccg gggctcccca ccacccccgc
     7921 agcggcgcta cgggcatacc atggtggcct ttgaccgcca cctctatgtg tttgggggtg
     7981 cggccgacaa cacgctgccc aacgagctgc actgctatga cgtggacttc cagacctggg
     8041 aggtcgtcca gcccagctcc gacagcgagg ttggtggggc tgaagtgccc gagcgagcct
     8101 gtgcttccga ggaggtgccc accctgacct atgaggagcg ggttggcttc aagaagtccc
     8161 gagatgtgtt tggcctggac tttggcacca cctcagccaa gcagcccacc cagcctgcct
     8221 cggagctgcc cagtgggagg ctcttccacg cagctgctgt catctcggac gccatgtaca
     8281 tcttcggggg cacggtggac aacaacatcc gcagcgggga gatgtacagg ttccagttct
     8341 cctgttaccc taaatgcacg ctgcacgagg actacgggcg gctgtgggag agccgccagt
     8401 tctgcgacgt ggagttcgtg ctgggtgaga aggaggagtg cgtgcagggc cacgtagcca
     8461 ttgtcacagc gcggagccgc tggcttcgca ggaagatcac gcaggcacgt gagaggctgg
     8521 cacagaagct ggagcaggag gcagctccag ttcccaggga ggcacccggc gtggctgctg
     8581 gtggggctag gccgcccctg ctgcacgtgg ctatcaggga ggccgaggca cgtcccttcg
     8641 aggtgctcat gcagttcctc tacaccgaca agatcaaata cccacggaaa ggccatgtgg
     8701 aggatgtgct gctcatcatg gatgtgtaca aactggcact gagcttccag ttgtgccgcc
     8761 tggagcagct gtgccgccag tacatcgagg cctccgtgga cctgcagaac gtgctggttg
     8821 tgtgcgagag tgccgcccgg ctgcagctga gccaactcaa ggagcactgc ctgaacttcg
     8881 tggtaaagga gtcccacttc aaccaggtga tcatgatgaa ggagttcgag cgcctctcct
     8941 ctccactgat agtggagatt gtgcggcgga agcaacaacc accccctcgc actcccttgg
     9001 accagccagt ggacattggc acatctctga tccaggacat gaaggcatac ctggagggag
     9061 caggtgcgga attctgtgac atcactctgt tgcttgacgg gcacccacgg ccagcccaca
     9121 aggctatcct ggccgctaga tccagctact ttgaagccat gttccggtcc ttcatgcccg
     9181 aagatgggca ggtgaacatc tccatcgggg agatggtgcc cagcaggcag gccttcgagt
     9241 ccatgctgcg ctacatctac tacggcgagg tcaacatgcc gcccgaggac tcgctctact
     9301 tgtttgcggc cccctactac tacggcttct acaacaaccg gctgcaggcg tactgcaagc
     9361 agaacctgga gatgaacgtg acggtgcaga acgtgctgca gatcctggag gcagctgaca
     9421 aaacgcaggc actggacatg aagcggcact gcctgcacat cattgtgcac cagttcacca
     9481 aggtctccaa gttgcccacc ctgcggtcgc tgagccagca gctgctgctg gacatcatag
     9541 actccctggc ctcccacatc tcagacaagc agtgcgcaga gctgggcgcc gacatctgag
     9601 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     9661 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     9721 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     9781 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     9841 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     9901 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     9961 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
    10021 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
    10081 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
    10141 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa
    10201 ttctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc
    10261 ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc
    10321 ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc
    10381 ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg aca
//
