LOCUS       sequence-345100-        7626 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses 3 overlapping genes (gfcB, gfcC, gfcD) from this one 
            operon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7626)
  AUTHORS   Larson MR, Biddle K, Gorman A, Boutom S, Rosenshine I, Saper MA
  TITLE     Escherichia coli O127 group 4 capsule proteins assemble at the outer
            membrane.
  JOURNAL   PLoS One. 2021 Nov 15;16(11):e0259900. doi: 
            10.1371/journal.pone.0259900. eCollection 2021.
  PUBMED    34780538
REFERENCE   2  (bases 1 to 7626)
  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: "PLoS One.";
            date: "2021-11-15"; volume: "16(11):e0259900. doi"; pages: " 
            10.1371/journal.pone.0259900. eCollection 2021"
COMMENT     Sequence Label: pSA10(gfcBCD)
FEATURES             Location/Qualifiers
     source          1..7626
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(38..55)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      208..294
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      386..413
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        432..523
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(524..592,593..1384)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTAGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(742..761)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1555..2143
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2044..2063
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2297..2314
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2329..2469
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2395..2414)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2555..2577
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        join(2802..2807,2808..2823,2824..2830)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2838..2854
                     /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     2843..2865
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     promoter        6428..6505
                     /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."
     CDS             6506..7588
                     /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     complement(6525..6544)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 agcttggctg ttttggcgga tgagagaaga ttttcagcct gatacagatt aaatcagaac
       61 gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg gtcccacctg
      121 accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc
      181 atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
      241 gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
      301 ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca
      361 taaactgcca ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt
      421 ctacaaactc ttttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
      481 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
      541 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
      601 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
      661 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
      721 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
      781 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
      841 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
      901 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
      961 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     1021 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     1081 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     1141 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     1201 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     1261 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacggcgg ggagtcaggc
     1321 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     1381 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     1441 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     1501 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     1561 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     1621 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     1681 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     1741 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     1801 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     1861 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     1921 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     1981 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     2041 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     2101 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     2161 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     2221 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     2281 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     2341 ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat
     2401 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     2461 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     2521 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     2581 tcaccgtcat caccgaaacg cgcgaggcag ccggatccgg gcttatcgac tgcacggtgc
     2641 accaatgctt ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa
     2701 atcactgcat aattcgtgtc gctcaaggcg cactcccgtt ctggataatg ttttttgcgc
     2761 cgacatcata acggttctgg caaatattct gaaatgagct gttgacaatt aatcatcggc
     2821 tcgtataatg tgtggaattg tgagcggata acaatttcac acaggaaaca gaattcccgg
     2881 gatgcgccct cttattttat cgattttcgc actatttctt gcgggatgta cgcacagcca
     2941 gcaaagtatg gtcgatacat ttcgcgccag cctattcgat aatcaggata tcaccgtagc
     3001 ggatcagcag atccaggcgt tgccttattc cacgatgtat ttacgcctta atgaagggca
     3061 acgaatcttt gtggtactgg gatatataga acaagaacaa agcaaatggt tatcccagga
     3121 taacgccatg ctggttaccc acaatggacg tcttttaaaa accgtcaaac ttaataataa
     3181 tctgctggaa gtgacaaatt ccgggcagga ccctctgcgt aacgcgctgg caataaaaga
     3241 cggcagccgc tggacgcgcg atattctctg gagcgaagac aaccattttc gctctgcgac
     3301 cctgagttcc actttttcct ttgctggatt agagacgctg aatattgcgg gtcgcgatgt
     3361 gctgtgtaat gtctggcagg aagaggtgac ttccacgctg ccagaaaaac agtggcaaaa
     3421 cacattctgg gtcgattcgg ctactggcca ggttcgtcaa agtcggcaaa tgttaggcgc
     3481 aggggttatt cccgtagaaa tgacgtttct taaacccgca ccatgaataa attacagtcg
     3541 tatttcattg ccagcgtcct ttacgtaatg acaccccatg cctttgcgca aggaatggtg
     3601 actatttatc tgcctggcga acaacagaca ctttccgttg gccccgtaga gaatgttgcc
     3661 cagttggtga cacaaccgca actgcgcgat cgcctctggt ggcctggtgc cttgctgacc
     3721 gactccgcag cgaaagctaa agcgcttaaa gactaccaac atgtcatggc acagcttgct
     3781 tcctgggaag cagaagctga cgacgatgtg gcggcaacga ttaaatccgt gcgccagcag
     3841 ttactcaatc tcaatatcac tggtcgctta ccggtcaaac tggaccctga ttttgtgcga
     3901 gtggatgaga acagcaatcc cccgctggtc ggggactata cgctctatac cgtacagcgt
     3961 ccggtgacta ttactctgct gggtgcggtt tctggggctg ggcaattgcc ctggctggca
     4021 ggccgtagcg ttaccgatta tctgcaagat cacccccgcc tcgcaggcgc ggacaaaaat
     4081 aacgttatgg tgattacgcc tgagggtgag actgtcgtcg cccctgttgc cctgtggaat
     4141 aaaagacatg tggaaccacc gccgggcagc cagctttggt tgggcttttc tgcgcatgta
     4201 ctgcccgaaa aatatgctga tttaaatgac cagattgtgt ctgttttaac gcaacgtgtt
     4261 cctgagtgat gaagaagaac tcttatcttt taagctgcct ggccattgcc gtctccagtg
     4321 cctgtcatgc tgaagtatta acctacccgg atccgctggg gtcgtcgcaa tcagactttg
     4381 gcggcacagg attgttgcag atgccaaatg cgcgcatcgc accggaaggt gaattcagcg
     4441 tcaactaccg ggataacgat caataccgtt tctactccac gtccgtggcg ctgttcccat
     4501 ggctggaagg caccattcgt tatacggatg tgcgcacacg caaatatagc cagtgggaag
     4561 atttcagcgg cgatcagtca tacaaagaca aatctttcga ttttaaactt cgcctgtggg
     4621 aagaaggtta cttgctaccg caagtggcgt ttggtaaacg tgatattgct ggtacgggtc
     4681 tgtttgacgg tgaatatttg gtggccagca agcaagcggg gccatttgat ttcaccctcg
     4741 ggatggcatg gggctacgcc ggtaatgcgg gcaatattac caacccgttt tgccgggtga
     4801 gcgataaata ttgtcatcgt gcagagtctc acgatgcggg cgatatcagc tttagcgata
     4861 tctttcgtgg cccggcttcc atctttggcg gcattgagta tcaaacgccg tggaatcccc
     4921 tgcgtctgaa actcgaatac gacggcaaca attaccagaa tgatttcgct ggcaaactgc
     4981 ctcaggcaag ccatttcaac gtcggtgcag tttatcgcgc tgccagctgg gcagacctca
     5041 acctcagtta tgaacgcggt aacacgttga tgtttggttt cactttacgg accaatttca
     5101 acgatctgcg ccctgccctg cgcgatacgc caaaaccggc atatcaacct gcgcctgaat
     5161 ctgaaggatt gcagtacacc acggtagcaa accaacttac cgccctgaag tataacgcgg
     5221 gctttgacgc gccagaaatt cagctgcgcg ataagacgct gtatatgtct ggtgagcaat
     5281 acaaataccg tgactctcgt gaagcggtcg atcgtgccaa ccggattctg gtgaataacc
     5341 tgccgcaagg cgttgagaag attagcgtga cgcaaaagcg cgagcatatg gcgatggtga
     5401 ctaccgaaac cgacgtagcc agcctgcgca aacagctggc cggtacagcg cctggtaaat
     5461 cagagcaact gcaacaacaa cgtgttgaag cagaagatct ttctgccttt ggtcggggct
     5521 accgtattcg tgaagatcgc tttagctact ctttcaaccc aacactttca cagtcgctgg
     5581 gcggcccgga agatttctat atgttccagc tggggctgat gtccagcgcc cgctactggt
     5641 ttaccgacca cctgctgctt gatggcggta ttttcaccaa tatttacaat aactacgaca
     5701 agtttaagtc ttcactatta cccgcggact ctaccctccc ccgcgtgcgc acgcatatcc
     5761 gtgactacgt tcgcaatgac gtttatctca acaacttgca ggcgaactat tttgccgact
     5821 taggcaatgg tttctatggc caggtgtatg gcggttatct ggaaacgatg tacgccggtg
     5881 tcggttccga gctgctttat cgcccgctag atgccagctg ggcgctgggt gtggacgtta
     5941 actacgtgaa gcaacgtgac tgggacaaca tgatgcgctt caccgattat tccacaccca
     6001 caggtttcgt gacggcttac tggaacccgc cgacgctcaa cggcgtactg atgaaactca
     6061 gcgttgggca atatctggca aaagataaag gcgcaacgat cgacgtcgcc aaacgctttg
     6121 acagcggcgt ggcggtaggg gtatgggcgg caatcagtaa cgtatctaaa gatgactacg
     6181 gcgaaggcgg ctttagtaaa ggtttttata tctcgattcc gttcgacttg atgaccattg
     6241 gacctaaccg caaccgcgcg gtggtttcgt ggacaccatt gacgcgtgat ggtggacaaa
     6301 tgctgtcacg caaataccag ctctatccaa tgacggcaga gcgagaagta ccggttggac
     6361 aatgagtcga cctgcagcca agcttcgaca attcgcgcgc gaaggcgaag cggcatgcat
     6421 ttacgttgac accatcgaat ggtgcaaaac ctttcgcggt atggcatgat agcgcccgga
     6481 agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta
     6541 tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc
     6601 gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt
     6661 ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc
     6721 cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc
     6781 cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca
     6841 caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga
     6901 tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga
     6961 ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga
     7021 gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt
     7081 ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc
     7141 gatagcggaa cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat
     7201 gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg
     7261 cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggatatct cggtagtggg
     7321 atacgacgat accgaagaca gctcatgtta tatcccgccg ttaaccacca tcaaacagga
     7381 ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc
     7441 ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc
     7501 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     7561 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt tagcgcgaat
     7621 tgtcga
//
