LOCUS       pETDuet1-HthermCCSbeta-HthermCCSalpha        7643 bp ds-DNA     circular SYN 11-FEB-2019
DEFINITION  LMBP 11134.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct

COMMENT     VNTDATE|869097600|
COMMENT     VNTDBDATE|869097600|
COMMENT     VNTNAME|pETDuet1-HthermCCSbeta-HthermCCSalpha|
FEATURES             Location/Qualifiers
     promoter        complement(1013..1090)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        1403..1421
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1422..1446
                     /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)."
     RBS             1461..1483
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2531..2548
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
     promoter        2623..2641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2642..2666
                     /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)."
     CDS             4011..4055
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
     terminator      4107..4154
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4191..4646
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(4761..5621)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
     promoter        complement(5622..5714)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5795..6383
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    6569..6708
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(6810..7001)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(7573..7643,1..1012))
                     /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"
ORIGIN
        1 aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca
       61 gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt
      121 gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt
      181 cttcggtatc gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc ccggactcgg
      241 taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa
      301 cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc
      361 cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca
      421 gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac
      481 ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac
      541 tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag
      601 cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc
      661 gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca
      721 tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt
      781 gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc
      841 ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca
      901 ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct
      961 gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca
     1021 ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt
     1081 cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag gaagcagccc
     1141 agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg
     1201 gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc
     1261 atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca
     1321 gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag
     1381 atcgatctcg atcccgcgaa attaatacga ctcactatag gggaattgtg agcggataac
     1441 aattcccctc tagaaataat tttgtttaac tttaagaagg agatatacca tggaacgtcg
     1501 ttggaagatc ggtaccccgt atctgaacga cagcacccgt atcattgtta tgggtattac
     1561 cggtcgtgag gcgagccagg tggttgcgga gagcgaagcg ctgtatccgg gttttgtggt
     1621 tgcgggtgtg accccgggta aaggtggcag cgaggttgcg ggtgtgccgg tttataacac
     1681 cgtgcgtgag gcgcaagaac gtcacccgga aatcaacacc ggcattgtgt atgttccgcc
     1741 ggcgagcgtg aaggatgcgg ttatcgagct gattgatgcg ggtatcggcg ttattttcat
     1801 cattaccgaa cacgtgccga tccgtgatac cgtttacttt tatcactacg cgaaagagcg
     1861 tggtaccatc attgtgggcc cgaccagcct gggttgcatc attccgaaga tcccggcgcg
     1921 tatcggtgcg attggtggca aagacccgag cgtggcgtat gcggatggtg gcctggttat
     1981 tctgagcaaa agcggtggcc tgaccaccac caccgcggaa atgttcaaac gtcgtggttg
     2041 gggcgtttac atggcgctgg cgctgggtgg cgacgtgatc agctgcacca cctttgcgga
     2101 tgcgattgag aacctggcgg acgatccgaa cgtgaagggc gttatcattc agggtgaagt
     2161 gggtggcagc tatgaggaac aagcggcgga gaccatcctg cgtctgtgga aggaaggtcg
     2221 ttggaacaag ccggtggcgg cgtttgttgc gggccgtttt caggagagcc tggaaggtgt
     2281 tagctttggt cacgcgggcg cgatcgtgga acgtggcaag ggcaaagcga ccgacaagat
     2341 ccgtgcgttc aacgaggttg gcaagattac cggcctggtg aaagttgcgg aattttatca
     2401 cgatctggtg cactgcatcg aggaactggg cgttccgcgt gacttcgagg atagcacccc
     2461 ggaaggcaag gtgaaaccgc tgtacagcac cattaacgag gaaaactgcc aatttaaagc
     2521 gggtggcagc catcaccacc accaccacta aaagcttgcg gccgcataat gcttaagtcg
     2581 aacagaaagt aatcgtattg tacacggccg cataatcgaa attaatacga ctcactatag
     2641 gggaattgtg agcggataac aattccccat cttagtatat tagttaagta taagaaggag
     2701 atatacatat gaacctgtac gagtatgaag cgtacgataa gatcttcaag aagtacggca
     2761 ttccgacccc ggaatacatg ttcgagagca gcgttagcga ccgtctggtt gaatttgtga
     2821 accagctggg cgagtgcgtg gttaaaagcc aagtgctggt tggtaaacgt ggcaaagcgg
     2881 gtgcggtgaa ggtttgcagc gatccgcaga gcgcgatcga aaccgcgcaa gcgctgctga
     2941 actacccggt ttatggcgag atgccggtgg gtgttctggt ggcgcgtaaa gtgaacatcc
     3001 tgaaggaact gtacgcgagc attacctata gcaccgaggt tcgtgcgccg gtgctgaccc
     3061 tgagcctgga aggtggcatg gacatcgagg aagttccgcc ggagaaagtg cgtagctgga
     3121 ccattaaccc gctgaagggt ctgtatccgc acatggtgcg taactacctg ctggaactgg
     3181 gcttcccgca ggagtatatg ggtatcctgc gtgaactgag cgaggtggtt agcaacatgt
     3241 accgtgcgtt ttgggaggcg gaagcgcgtc tgctggaaat caacccgctg gcgatttgcg
     3301 atgttaacgg caagctgaaa gtgtatgcgc tggacgcggt ggttaccatc gacgatgacg
     3361 cgagcgttcc gccgagcaaa atttacggtg tgcgtaccgc gatgaagcgt ccgccgaccg
     3421 agcgtgaaat cgaggcgagc ctgattgatc gtgatgacca ccgtggcaag gcgggtagct
     3481 atgttgaagt ggatggcgac atcgcgatga tgaccttcgg tggcggtggc agcaccgtta
     3541 ccatcgagac cacctatgcg attggtctga aaccggcgaa ctttaccgat attggtggca
     3601 acccgccggc ggagaagatg tacaaaatca ccaagatcat tctgagcaag ccgggcattc
     3661 gtggtgttct ggtgtgcggt ggcaccgcga acaacacccg tatcgacgtt accctgggtg
     3721 aaggcgtggc gaacgcgatt cgtgatctgt acaaagaggg caagctgaac ccggactgga
     3781 tctgggtggt tcgtcgtaac ggcccggaag cggagaaagg tctgcgtatg ctgtatgaag
     3841 cgtttaagga gtgcaaggtg aaaggtgaaa tttacgatag cagcctgccg ctgaccgaag
     3901 cgccgatccg tctgaaagag ctgctggata tttgcaccag cgcgcagagc gaggaccgtc
     3961 acctgaccga ggaacaagcg aaggacatgg gtatctaact cgagtctggt aaagaaaccg
     4021 ctgctgcgaa atttgaacgc cagcacatgg actcgtctac tagcgcagct taattaacct
     4081 aggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
     4141 tgaggggttt tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg
     4201 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4261 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4321 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4381 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     4441 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4501 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4561 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     4621 taacaaaata ttaacgttta caatttctgg cggcacgatg gcatgagatt atcaaaaagg
     4681 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4741 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4801 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4861 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4921 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4981 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5041 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5101 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5161 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5221 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5281 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5341 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5401 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5461 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5521 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5581 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatca
     5641 tgattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5701 gaaaaataaa caaataggtc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5761 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5821 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5881 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5941 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6001 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6061 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6121 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6181 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6241 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6301 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6361 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6421 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6481 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     6541 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     6601 gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     6661 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
     6721 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     6781 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     6841 gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc
     6901 tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata
     6961 aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg
     7021 ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg
     7081 gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta
     7141 tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca
     7201 gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg
     7261 gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt
     7321 catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt
     7381 atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac
     7441 gacaggagca cgatcatgct agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     7501 aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta
     7561 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     7621 tgaatcggcc aacgcgcggg gag
//
