LOCUS       sequence-183583-        7240 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of H. influenzae MolBC (ABC Transporter) in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7240)
  AUTHORS   Pinkett HW, Lee AT, Lum P, Locher KP, Rees DC
  TITLE     An inward-facing conformation of a putative metal-chelate-type ABC 
            transporter.
  JOURNAL   Science. 2007 Jan 19;315(5810):373-7. Epub 2006 Dec 7.
  PUBMED    17158291
REFERENCE   2  (bases 1 to 7240)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science."; 
            date: "2007-01-19"; volume: "315(5810)"; pages: "373-7. Epub 2006 
            Dec 7"
COMMENT     Sequence Label: MolBC
FEATURES             Location/Qualifiers
     source          1..7240
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      6..594
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     495..514
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     748..765
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    780..919
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(845..864)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1005..1027
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1021..1212)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2021..3103)
                     /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"
     primer_bind     3065..3084
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3104..3181)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3387..3406
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3490..3509
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3490..3508
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3509..3533
                     /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             3548..3570
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3583..3606
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             3622..3636
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     RBS             4666..4688
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5461..5478
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5531..5549)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5545..5592
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      5629..6084
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5716..5735)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5926..5947
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6110..6214
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6215..6283,6284..7075)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6433..6452)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
       61 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
      121 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
      181 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
      241 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
      301 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
      361 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
      421 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
      481 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
      541 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
      601 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
      661 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
      721 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
      781 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc
      841 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg
      901 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
      961 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     1021 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc
     1081 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt
     1141 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc
     1201 ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc
     1261 gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact
     1321 ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac
     1381 tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca
     1441 gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag
     1501 actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt
     1561 gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag
     1621 gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg
     1681 ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt
     1741 gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat
     1801 cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg
     1861 tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg
     1921 cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg
     1981 tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc
     2041 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     2101 gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc
     2161 ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca
     2221 ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat
     2281 cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc
     2341 gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct
     2401 cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt
     2461 ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac
     2521 gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga
     2581 ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg
     2641 gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag
     2701 caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga
     2761 gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca
     2821 ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg
     2881 cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt
     2941 gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc
     3001 gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga
     3061 caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt
     3121 gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt
     3181 ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg
     3241 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac
     3301 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     3361 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     3421 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc
     3481 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
     3541 aaataatttt gtttaacttt aagaaggaga tataccatgg gccatcatca tcatcatcat
     3601 catcacagca gcggccatat cgacgacgac gacaagcata tgcagccaga tagttaccct
     3661 aaaattctct ttggcttaac gctgttgctt gtgattaccg ccgtgatttc tttgggaata
     3721 gggcggtatt ctttgtctgt accgcaaatt ggtcaaattc tgtgggcaaa agcgaccgca
     3781 cttgagatcg atccagtgca gcaacaggtg atttttcaag tgcgtttacc tcgaatttta
     3841 accgcacttt gcgtgggtgc cggtttggca ttgagtggtg tggtattgca aggcattttt
     3901 cgtaacccgt tagtgaatcc gcatattatc ggcgtgacct ccggttcagc cttcggtggc
     3961 accttggcaa ttttctttgg attcagttta tacggcttat ttaccagcac tattttattc
     4021 ggcttcggca cgcttgcctt ggtttttcta tttagcttca aatttaatca acgcagcttg
     4081 ttgatgctta ttcttatcgg catgatttta agcggacttt tttcagccct agtgagccta
     4141 ttgcaataca tttccgatac agaagaaaaa ttgccaagca ttgtgttttg gttgatgggg
     4201 agttttgcca cctctaactg ggaaaaactc ctctttttct ttgtcccgtt tttgctgtgt
     4261 agctcgattt tgctcagctt aagttggcgc ttgaatttac tgtctttaga tgagaaagaa
     4321 gccaaggctt tgggcgtgaa aatggcccca ttacgttggt tagtgatctt tttaagcggt
     4381 tcgttagtgg catgccaggt ggcaattagc ggcagtatcg gctgggtcgg gctgatcatt
     4441 cctcatctta gccgtatgct ggtaggtgcc aatcatcaaa gtttattgcc ttgcaccatg
     4501 cttgttgggg cgacttatat gttgcttgtg gacaatgttg cgcgttccct atccgatgcc
     4561 gaaattccaa tcagtatttt gaccgcactt attggtgcac cgttgtttgg tgtattggta
     4621 tataaactaa aacgcggagg catgaatgaa taatctagaa ataattttgt ttaactttaa
     4681 gaaggagata tacatatgaa taaagcgctc tccgtggaaa atctaggttt ttattaccaa
     4741 gcggaaaact tcctgtttca gcaactgaat tttgatttga ataaaggaga tattctggcg
     4801 gttttagggc aaaacggttg cgggaaaagt accttgttgg atttgttgct tggcattcat
     4861 cgcccgattc agggcaagat tgaggtgtat caatctatcg gttttgtgcc gcaattcttt
     4921 tcttcgcctt ttgcttattc cgtattggat atcgtgttga tggggcgctc aacccatatt
     4981 aatacttttg ccaaaccgaa atcgcatgat tatcaagtcg ccatgcaggc actggattat
     5041 ttgaacttga cccatttggc aaagcgcgaa tttacttcgc tttcgggcgg gcagcgacaa
     5101 ctaattttaa ttgcgagagc cattgcttca gaatgcaaat tgatactgtt ggatgagcca
     5161 acttcagcat tagatcttgc aaatcaagat attgtactgt ctttacttat cgatttggca
     5221 caatcacaga acatgactgt cgtattcaca acacatcaac ctaatcaagt tgtagcgata
     5281 gccaataaaa ctttattgtt aaataaacaa aattttaaat ttggcgaaac aaggaatatt
     5341 ttaacctcag aaaatttgac cgcacttttt catttaccga tgtttgaaca acaagctcaa
     5401 tataaagaat ctttttttac gcattttgtt cccttgtata aaacgttatt aaagtaagag
     5461 caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg
     5521 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     5581 aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta
     5641 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     5701 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     5761 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     5821 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     5881 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     5941 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     6001 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     6061 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     6121 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     6181 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     6241 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     6301 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     6361 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     6421 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     6481 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     6541 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     6601 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     6661 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     6721 agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg
     6781 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     6841 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     6901 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     6961 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     7021 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     7081 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     7141 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     7201 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
//
