LOCUS       pTrc-hACLmut        7644 bp ds-DNA     circular SYN 11-FEB-2019
DEFINITION  LMBP 11127.
SOURCE
  ORGANISM

COMMENT     VNTDATE|869097600|
COMMENT     VNTDBDATE|869097600|
COMMENT     VNTNAME|pTrc-hACLmut|
FEATURES             Location/Qualifiers
     terminator      60..146
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(372..389)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
     CDS             complement(405..434)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
     regulatory      2893..2902
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             complement(3735..3761)
                     /codon_start=1
                     /product="synthetic cistron containing a ribosome binding 
                     site (Shine-Dalgarno sequence), for enhancing the bacterial
                     expression of a downstream cistron (Schoner, 1997)"
                     /label=minicistron
                     /note="This first cistron should terminate 3 bp upstream of
                     the ATG for the second cistron."
     misc_feature    3812..3881
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region 
                     (Berg et al., 1989)"
     protein_bind    3898..3914
                     /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)."
     promoter        complement(3922..3951)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             complement(4183..5265)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(5266..5343)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     misc_feature    5529..5669
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5855..6443)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6614..7474)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
     promoter        complement(7475..7566)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      7585..7612
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cgtcctgccc gccaccctcc gggccgttgc ttcgcaacgt tcaaatccgc tcccggcgga
       61 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
      121 ttcgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg catggggaga
      181 ccccacacta ccatcggcgc tacggcgttt cacttctgag ttcggcatgg ggtcaggtgg
      241 gaccaccgcg ctactgccgc cagacaaatt ctgttttatc agaccgcttc tgcgttctga
      301 tttaatctgt atcaggctga aaatcttctc tcatccgcca aaacagccaa gctggagacc
      361 gtttaaactc aatgatgatg atgatgatgg tcgacggcgc tattcagatc ctcttctgag
      421 atgagttttt gttctacgta agcttcgaat tcgcccttca tgctcatgtg ttccggaaga
      481 acatatgaaa tatcatccca cggatgacga tacagcccct gcttcagcct cttctgatca
      541 agatagtgtc caatgaaccc catactcctt cccagcacaa agatgccatt gagggctcca
      601 atgtcaatat attcatcagc ttcctcccga gtaaaggacc cacagtttct aagcatgtct
      661 acaaatgcga ctccgatgag accatctaca ttcaggataa gatttggctt cttcgaggtg
      721 gtaatcttct ctacttccag tgcataatcg agcagaggag tggcagggaa gtgctgcctg
      781 acgtaatctt tgaggatctg cactcgcatg tctgggttgt ttatcgactt cactcggtga
      841 ccaatgccca tgatcagctt cccttccttc ttcatcttgt tcacaaactc catggggata
      901 atgccactgt caaaggcttt actgaacatc ttggctgctg catccaaggc acccccaaac
      961 cgatccccga tggtgagcag ccccgaggtg aggctggaga ccaggtcttt cccagctcgc
     1021 gcacaaatga tggtgttgtg ggctccagag acggctggcc cgtgatcagc tgtcaccatc
     1081 agacacatct caatgaactg gcaagagtac ttaggcaacc ttttctggaa ccagaggagg
     1141 ccgaggaccc cgccaatgcc catctcttcc ttgaagacct cagtgatggg catgcccgcg
     1201 tagatgagct cctgtcctcg ctcatcgcag atgctggtca tgaacgaggc aggtttgcgg
     1261 atcaaaccaa gctccctggc ccaggagtag tccatgggca cggttggggg cggcacctcc
     1321 tgggcaggta caatgactcc attggccacg agatcttcgt atacagactg gatgatctct
     1381 ccaagctcat caaagctccg gggcacaaac actcctgctt ccttcaaagc ctggttcttg
     1441 gctactgcag tttcagaagc ctggttggca caagctccag catggccaaa ctggacctca
     1501 gaggagaaca tggtggcaca cgtcccgatg caccagcaga cgatgggctt agtgaggcgg
     1561 ccctccttga tgccccggca aatcttatat tcctcagtgc ccccaatctc tccaagaacc
     1621 acaatcattt tgactcctgg agtgtcctga tagcgtaaca catgatccat gaatgtggag
     1681 cccgggtacc tgtccccacc aatggccacg ccatcataga cgccatccgt ggtccgagag
     1741 atgatattgt tgagctcgtt ggacatgcct ccggaacgtg agacataggc cacgctgcct
     1801 gggcggtaca gtttggaggc caggatgttg tccagcatcc cacctgtgtt gccaatctta
     1861 aagcacccag gcttgatgcc tccaacagtg gcaggtccga tgatggtcac tcccttctgg
     1921 tccgccttct tgatcagctt tctcgtgagg gcctcaggga tgccttcagc tatgatggcg
     1981 atggtccgga tctgggcata gttcatggtc tccatggtgc tgtcataggc agagcggaga
     2041 gaggcaaagt tgatgagcac atctacctcc gggtgcttcc tcatggcatc agccatgttc
     2101 ttgaagacag ggatcaggat ctctttgtgc ccccagtaaa acttctgctt gtggtcccca
     2161 gtgaaagggt agaccatggc agccactgag ggctcgtctc gggagcagac atagtcaaag
     2221 tccagcatgc cttgcacggc ccgggtctgc atgccccaca caatggcctt ggtgtggcgg
     2281 ctgaagaggg tggtgctctt tccttgtggc atggcaggct tggccttctt tgcaggcgcc
     2341 acctcatcgg ccctggactc agaaaaagat gctgtcctgc tgggggctgg cgtcgatgtg
     2401 ctcccgctgg cgttgaggag gaagtttgca gtgtgggccg ctgtgggtgg ctggttgggg
     2461 atgggccggt ggcccagggc catgcccaca atggccgtca tgtgagtctc tgtgccaaag
     2521 acatggatgg ggatcccagt ggtcttcccg acttctccca tcacccgtaa gccctcctga
     2581 tagttggggc cacctcttcg gacaaagatt gtgacttcgt gctccttcag ggggccctgg
     2641 taatctcgaa ttgctctcac gatgcccttg aacgtggcag ccacgttggt gaagtttgcg
     2701 atgctgcctc caatgatgag gatcttgcca tctgggtgct tctctcgggt catgagggag
     2761 aggatagtct tggcatagtc ataggtctgc tgctcgctgg gggcgcctga gtactcccca
     2821 tagtttgcca gctcgttgac accccctaga tcacagatgg tatcgctgta cacgacagag
     2881 gcgccacccc cggccaccat ggtccagatc ctccctttgg ggttcagcaa ggtcagcttc
     2941 aggcttgccc cacttttggc atcgaggtct gcaatgtagg cttcctctgg atatgcctcc
     3001 cgcccgaagg ggggagggaa ctcgatgtca ccccacttca ctttgcagat gtagtcggca
     3061 gtggcgtcca ccttggccgc caagtcaagg acatagactc catctttggt cactacaagg
     3121 ggattgatct cgaggtaggt gaagtacaag tcctcgtaga aattgaagag gccggagata
     3181 aaactggcca gaatttcttt cttgtcttca ggggcgtgga ccaacaggtg ttttttgatg
     3241 tcctcaggat tcagtttctc atccacgcca acaagcagct tctgggcctt ggcgtccaca
     3301 tcacccacgt ccacaccccc ctcgtggtgg aacaggacgt agtccccttc tcgggtggca
     3361 tagatgcaga catagaactc ctcagcctga ctgtggggga cgaagggctc gatcagaaag
     3421 ttcttgagga agcctgtggc cttgccaact gtggcttcct gtcccagccg tggcttcagc
     3481 caggacttga ccccatccag agtgaggttg accccaacga gaccaagttt tccacgacgt
     3541 ttgatcagct ggtctggctt gactaccaag ttctggctga gcagccaggg gtggtcctgc
     3601 agcaagcggg cccagtctgt gtcaggagtg acccgagcat acttgaaccg attctggatg
     3661 gctgaggtgg tacagatgaa cttgtaaagg agttctttgc ccgtctgctc tgaaattgcc
     3721 ttggccgaca tggtttattc ctccttattt aatcgataca ttaatatata cctctttaat
     3781 ttttaataat aaagttaatc gataattccg gtcgagtgcc cacacagatt gtctgataaa
     3841 ttgttaaaga gcagtgccgc ttcgcttttt ctcagcggcg ctgtttcctg tgtgaaattg
     3901 ttatccgctc acaattccac acattatacg agccggatga ttaattgtca acagctcatt
     3961 tcagaatatt tgccagaacc gttatgatgt cggcgcaaaa aacattatcc agaacgggag
     4021 tgcgccttga gcgacacgaa ttatgcagtg atttacgacc tgcacagcca taccacagct
     4081 tccgatggct gcctgacgcc agaagcattg gtgcaccgtg cagtcgatga taagctgtca
     4141 aaccagatca attcgcgcta acttacatta attgcgttgc gctcactgcc cgctttccag
     4201 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4261 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt
     4321 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
     4381 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
     4441 atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc
     4501 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
     4561 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     4621 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
     4681 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     4741 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
     4801 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     4861 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     4921 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     4981 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     5041 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     5101 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     5161 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     5221 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
     5281 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcacc attcgatggt
     5341 gtcaacgtaa atgcatgccg cttcgccttc gcgcgcgaat tgatctgctg cctcgcgcgt
     5401 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     5461 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     5521 tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtatac tggcttaact
     5581 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     5641 gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc
     5701 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5761 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5821 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5881 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5941 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6001 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6061 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6121 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6181 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6241 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     6301 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6361 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6421 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6481 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6541 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6601 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6661 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6721 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6781 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6841 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6901 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6961 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7021 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7081 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7141 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7201 ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa
     7261 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7321 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7381 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7441 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7501 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7561 aacaaaagag tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttaatt
     7621 tgatgcctgg cagtttatgg cggg
//
