LOCUS       pET11a-MconcACL        8820 bp ds-DNA     circular SYN 11-FEB-2019
DEFINITION  LMBP 11132.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct

COMMENT     VNTDATE|869097600|
COMMENT     VNTDBDATE|869097600|
COMMENT     VNTNAME|pET11a-MconcACL|
FEATURES             Location/Qualifiers
     CDS             1098..1115
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
     terminator      1183..1230
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(1405..1433)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     promoter        1546..1650
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1651..2511
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
     rep_origin      2682..3270
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3456..3595
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3697..3888)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5200..6282)
                     /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(6283..6360)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        6669..6687
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6688..6712
                     /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             6727..6749
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 ccggtacctg cagcccgtac ctgccggcga gcgtgcaatt tggtcacgcg ggcgcgaaag
       61 cgaacaccga aaaggagacc gcgcaggcga agaacgatgc gttccgtcaa gcgggtgcgt
      121 acgttccgcg tagctttgac gattatggcg aaatggtgcg tcaggtttac gacatgctgc
      181 tgacccgtgg tattgtgcaa aaattcgatg aaccggaagt gccgcgtatc ccgaccgact
      241 atagcaaagc gctggcgacc ggcgatatcc gtaagccgac cacctttatc tgcaccatta
      301 gcgacgatag cggcgaggaa ctgctgtacg cgggcaagaa actgagcgac gtgctggatc
      361 gtaaaatggg tattggtggc gttatcggcc tgctgtggtt caagaaagaa ctgccggagt
      421 acgcggcgca ctttatcgag ctggtgatcc agattgttgc ggaccacggt ccggctgtga
      481 gcggtgcgca caacgcgatt gttgcgagct gcgcgggtaa agatctgatc agcagcctgt
      541 gcagcggtct gctgaccatt ggcccgcgtt ttggtggcgc gattgatgat gcggcgcgtg
      601 agtttaaacg tgcgcaggaa accggtctgg cgccggagca attcgtgggt gaaatgaaga
      661 aaaagggcat caacattccg ggtattggcc acaaaatcaa gagcgtgaaa aacccggaca
      721 agcgtgttca gctgctgatc agctatgcgc gtgcgaactt tccgagcacc gaactgctga
      781 actacgcgct gcaagtggag gaactgacca ccgcgaaaaa gggtaacctg attctgaacg
      841 ttgacggttg catcggcatt ctgttcatcg atctgatgag cagctgcggc gcgtttagca
      901 aagaggagat cgacgaggtg gttcgtctgg gttacctgaa cggcctgttc gcgctgggtc
      961 gtagcatcgg tctgattggc cacatcctgg accagaaacg tctgggcagc cgtctgtatc
     1021 gtcacccggc ggaagatatt gcgtacatga tgccgagcga ggaagagatc caatgcaagc
     1081 gtgatcgtgg tggcagccac caccaccacc accactaagg atccggctgc taacaaagcc
     1141 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     1201 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatatccc
     1261 gcaagaggcc cggcagtacc ggcataacca agcctatgcc tacagcatcc agggtgacgg
     1321 tgccgaggat gacgatgagc gcattgttag atttcataca cggtgcctga ctgcgttagc
     1381 aatttaactg tgataaacta ccgcattaaa gcttatcgat gataagctgt caaacatgag
     1441 aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     1501 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     1561 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     1621 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     1681 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     1741 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     1801 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     1861 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     1921 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     1981 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     2041 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     2101 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     2161 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     2221 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     2281 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     2341 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     2401 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     2461 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     2521 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     2581 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     2641 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2701 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2761 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2821 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2881 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2941 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     3001 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3061 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3121 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3181 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3241 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3301 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     3361 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     3421 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     3481 catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg
     3541 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
     3601 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     3661 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     3721 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     3781 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     3841 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt
     3901 gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac
     3961 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
     4021 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt
     4081 taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa
     4141 cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa
     4201 gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg
     4261 ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt
     4321 cctcaacgac aggagcacga tcatgcgcac ccgtggccag gacccaacgc tgcccgagat
     4381 gcgccgcgtg cggctgctgg agatggcgga cgcgatggat atgttctgcc aagggttggt
     4441 ttgcgcattc acagttctcc gcaagaattg attggctcca attcttggag tggtgaatcc
     4501 gttagcgagg tgccgccggc ttccattcag gtcgaggtgg cccggctcca tgcaccgcga
     4561 cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta caatccatgc caacccgttc
     4621 catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca gcggtccaat gatcgaagtt
     4681 aggctggtaa gagccgcgag cgatccttga agctgtccct gatggtcgtc atctacctgc
     4741 ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag aagaatcata
     4801 atggggaagg ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg
     4861 gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg
     4921 acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc
     4981 gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt
     5041 cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc
     5101 gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt
     5161 gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5221 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5281 cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc
     5341 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag
     5401 gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc
     5461 gtcgtatccc actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg
     5521 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc
     5581 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc
     5641 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg
     5701 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
     5761 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
     5821 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
     5881 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
     5941 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
     6001 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
     6061 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
     6121 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
     6181 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
     6241 accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg
     6301 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc
     6361 cgggatctcg acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt
     6421 tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca
     6481 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
     6541 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
     6601 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc
     6661 cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga
     6721 aataattttg tttaacttta agaaggagat atacatatgg cgcaacgtgg tatccgtgag
     6781 tacgacgcga agaacctgct ggcgcgttac ctgccggaat atctggacga tttcagctat
     6841 aaaggtaacc tggcgctggt tggtccggag accgacattg aaggcctgga ggcggaaaac
     6901 ccgtggctga agaccacccg tctggtggtt aaaccggatc agctgtttgg caagcgtggt
     6961 aaactgggcc tggtgctgct ggacgcggat tgggaggaag cgaaggagta cctgaacgaa
     7021 aaaatgggtc tggaagtgac catcggtggc attaccggcc gtctgagcta tttcctgatc
     7081 gaaccgttta ccccgcacaa agaggagtac tatgtggcga tcagcagcga ctacgagggt
     7141 gataacattt tctttagcat ggacggtggc gtgggcgttg aggaaaactg ggagaaagtg
     7201 atcagcattc acgttgatag cctggaaggt atcgatgcgc tggatgtggg cagcaagctg
     7261 ccggcggagc tgggtgacaa acgtgcgctg gttgaggaat tcattaccgc gctgtggcgt
     7321 ttctacagcg ataccggctt tgcgtatgtt gaaatcaacc cgttcacctt tagcggccgt
     7381 ggtattgtgc cgctggacat ggttgcgaag ctggacgatg cggaggaata ctggcaaaag
     7441 aaacgttgga gcgagctggc gttcccggaa ccgtttggtc gtaccccgag caaagaggag
     7501 ctgtttatca aggagattga tagcaaaacc ggtgcgagcc tgaaactgac catcctgaac
     7561 ccggaaggtc gtgtttggac gatggtggcg ggtggcggtg cgagcgttat ctatgcggac
     7621 accatttgcg atctgggtca cgcggacgag atggcgaact acggtgaata tagcggcgat
     7681 ccgaacaccg aggaaaccta ccactatacc tgcaccattc tggacctgat gacccgtagc
     7741 aagaacccga acggtaaagt gctgctgatc ggcggtgcga ttgcgaactt caccgatgtt
     7801 gcgaagacct ttaaaggcgt ggttatggcg ctggaggaat accagcaaaa actgcaggaa
     7861 gcggacatcg aaatttacgt gcgtcgtggc ggtccgaact atgagcaagg tctgaagctg
     7921 atgcgtgatc tgggtaaacg tctgggcgtg ccgatccagg ttcacggccc ggaaacccac
     7981 atgacccgta ttgttccgct ggcgctggag gaaaaccagt aagcattcga aggagatatc
     8041 acatgagccg taaagactat gtgctgttcg atattaacac caaggcgttt gtttacggtt
     8101 atcagaccaa cgcgatccaa cgtatgctgg acttcgatta cgtgtgcaaa cgtagcagcc
     8161 cgagcattag cgcgatcatt aacccgagcc gtgcgggtat ccacaaagcg ttttggggca
     8221 ccaaggagat cattctgccg atgtataaga ccatcccgct ggcggcgctg gcgtacccgg
     8281 aagcggacgt gatggttaac ttcgcgagcc accgtagcgc gtttgaaacc acgatggagg
     8341 cgctgaaaga agataccatc cgtattgtgg cggttattgc ggagggtgtg ccggaacgtc
     8401 agagccgtgt tatggcggcg accgcgcgta aactggacaa gatcgtgatt ggtccggcga
     8461 ccgttggtgg catgaccgcg ggtgcgttcc gtattggtaa caccgcgggc accatcgaga
     8521 acatcattgc gagcaaactg tatcgtccgg gttgcgtggg cttcgttagc aagagcggtg
     8581 gcatgctgaa cgaggcgttt aacatcatta gccgtaacag cgacggtatt tatgaaggcg
     8641 ttgcgatcgg tggcgaccgt tacccgggta gcaacatgct ggatcacatt ctgcgttatg
     8701 agcgtaaccc ggcgatcaag atgattgcgt gcctgggcga actgggtggc gaagatgagt
     8761 acatgatcat tcaggcgctg aaagagaaga aaatcaccaa gccgctggtt gcgtgggtta
//
