LOCUS       sequence-345140-        7644 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses 3 overlapping genes (gfcB-6xHis, gfcC, gfcD) from this one
            operon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7644)
  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 7644)
  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(gfcB-His-CD)
FEATURES             Location/Qualifiers
     source          1..7644
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(56..142)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     295..312
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(388..1470)
                     /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     1432..1451
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1471..1548)
                     /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             complement(4456..4473)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5129..5151)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5140..5156
                     /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(join(5164..5170,5171..5186,5187..5192))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     primer_bind     complement(5417..5439)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5525..5665
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5580..5599
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5680..5697)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5851..6439)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5931..5950)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6610..7401,7402..7470))
                     /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     7233..7252
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7471..7562)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(7581..7608)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ctgcccgcca ccctccgggc cgttgcttcg caacgttcaa atccgctccc ggcggatttg
       61 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtctttcg
      121 actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcatg gggagacccc
      181 acactaccat cggcgctacg gcgtttcact tctgagttcg gcatggggtc aggtgggacc
      241 accgcgctac tgccgccagg caaattctgt tttatcagac cgcttctgcg ttctgattta
      301 atctgtatca ggctgaaaat cttctctcat ccgccaaaac agccaagctt cgacaattcg
      361 cgctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
      421 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
      481 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
      541 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
      601 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
      661 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
      721 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
      781 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
      841 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
      901 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
      961 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     1021 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     1081 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     1141 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     1201 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     1261 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     1321 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     1381 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     1441 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     1501 gcgctatcat gccataccgc gaaaggtttt gcaccattcg atggtgtcaa cgtaaatgca
     1561 tgccgcttcg ccttcgcgcg cgaattgtcg aagcttggct gcaggtcgac tcattgtcca
     1621 accggtactt ctcgctctgc cgtcattgga tagagctggt atttgcgtga cagcatttgt
     1681 ccaccatcac gcgtcaatgg tgtccacgaa accaccgcgc ggttgcggtt aggtccaatg
     1741 gtcatcaagt cgaacggaat cgagatataa aaacctttac taaagccgcc ttcgccgtag
     1801 tcatctttag atacgttact gattgccgcc cataccccta ccgccacgcc gctgtcaaag
     1861 cgtttggcga cgtcgatcgt tgcgccttta tcttttgcca gatattgccc aacgctgagt
     1921 ttcatcagta cgccgttgag cgtcggcggg ttccagtaag ccgtcacgaa acctgtgggt
     1981 gtggaataat cggtgaagcg catcatgttg tcccagtcac gttgcttcac gtagttaacg
     2041 tccacaccca gcgcccagct ggcatctagc gggcgataaa gcagctcgga accgacaccg
     2101 gcgtacatcg tttccagata accgccatac acctggccat agaaaccatt gcctaagtcg
     2161 gcaaaatagt tcgcctgcaa gttgttgaga taaacgtcat tgcgaacgta gtcacggata
     2221 tgcgtgcgca cgcgggggag ggtagagtcc gcgggtaata gtgaagactt aaacttgtcg
     2281 tagttattgt aaatattggt gaaaataccg ccatcaagca gcaggtggtc ggtaaaccag
     2341 tagcgggcgc tggacatcag ccccagctgg aacatataga aatcttccgg gccgcccagc
     2401 gactgtgaaa gtgttgggtt gaaagagtag ctaaagcgat cttcacgaat acggtagccc
     2461 cgaccaaagg cagaaagatc ttctgcttca acacgttgtt gttgcagttg ctctgattta
     2521 ccaggcgctg taccggccag ctgtttgcgc aggctggcta cgtcggtttc ggtagtcacc
     2581 atcgccatat gctcgcgctt ttgcgtcacg ctaatcttct caacgccttg cggcaggtta
     2641 ttcaccagaa tccggttggc acgatcgacc gcttcacgag agtcacggta tttgtattgc
     2701 tcaccagaca tatacagcgt cttatcgcgc agctgaattt ctggcgcgtc aaagcccgcg
     2761 ttatacttca gggcggtaag ttggtttgct accgtggtgt actgcaatcc ttcagattca
     2821 ggcgcaggtt gatatgccgg ttttggcgta tcgcgcaggg cagggcgcag atcgttgaaa
     2881 ttggtccgta aagtgaaacc aaacatcaac gtgttaccgc gttcataact gaggttgagg
     2941 tctgcccagc tggcagcgcg ataaactgca ccgacgttga aatggcttgc ctgaggcagt
     3001 ttgccagcga aatcattctg gtaattgttg ccgtcgtatt cgagtttcag acgcagggga
     3061 ttccacggcg tttgatactc aatgccgcca aagatggaag ccgggccacg aaagatatcg
     3121 ctaaagctga tatcgcccgc atcgtgagac tctgcacgat gacaatattt atcgctcacc
     3181 cggcaaaacg ggttggtaat attgcccgca ttaccggcgt agccccatgc catcccgagg
     3241 gtgaaatcaa atggccccgc ttgcttgctg gccaccaaat attcaccgtc aaacagaccc
     3301 gtaccagcaa tatcacgttt accaaacgcc acttgcggta gcaagtaacc ttcttcccac
     3361 aggcgaagtt taaaatcgaa agatttgtct ttgtatgact gatcgccgct gaaatcttcc
     3421 cactggctat atttgcgtgt gcgcacatcc gtataacgaa tggtgccttc cagccatggg
     3481 aacagcgcca cggacgtgga gtagaaacgg tattgatcgt tatcccggta gttgacgctg
     3541 aattcacctt ccggtgcgat gcgcgcattt ggcatctgca acaatcctgt gccgccaaag
     3601 tctgattgcg acgaccccag cggatccggg taggttaata cttcagcatg acaggcactg
     3661 gagacggcaa tggccaggca gcttaaaaga taagagttct tcttcatcac tcaggaacac
     3721 gttgcgttaa aacagacaca atctggtcat ttaaatcagc atatttttcg ggcagtacat
     3781 gcgcagaaaa gcccaaccaa agctggctgc ccggcggtgg ttccacatgt cttttattcc
     3841 acagggcaac aggggcgacg acagtctcac cctcaggcgt aatcaccata acgttatttt
     3901 tgtccgcgcc tgcgaggcgg gggtgatctt gcagataatc ggtaacgcta cggcctgcca
     3961 gccagggcaa ttgcccagcc ccagaaaccg cacccagcag agtaatagtc accggacgct
     4021 gtacggtata gagcgtatag tccccgacca gcgggggatt gctgttctca tccactcgca
     4081 caaaatcagg gtccagtttg accggtaagc gaccagtgat attgagattg agtaactgct
     4141 ggcgcacgga tttaatcgtt gccgccacat cgtcgtcagc ttctgcttcc caggaagcaa
     4201 gctgtgccat gacatgttgg tagtctttaa gcgctttagc tttcgctgcg gagtcggtca
     4261 gcaaggcacc aggccaccag aggcgatcgc gcagttgcgg ttgtgtcacc aactgggcaa
     4321 cattctctac ggggccaacg gaaagtgtct gttgttcgcc aggcagataa atagtcacca
     4381 ttccttgcgc aaaggcatgg ggtgtcatta cgtaaaggac gctggcaatg aaatacgact
     4441 gtaatttatt catgggtggt gatgatggtg atgtgcgggt ttaagaaacg tcatttctac
     4501 gggaataacc cctgcgccta acatttgccg actttgacga acctggccag tagccgaatc
     4561 gacccagaat gtgttttgcc actgtttttc tggcagcgtg gaagtcacct cttcctgcca
     4621 gacattacac agcacatcgc gacccgcaat attcagcgtc tctaatccag caaaggaaaa
     4681 agtggaactc agggtcgcag agcgaaaatg gttgtcttcg ctccagagaa tatcgcgcgt
     4741 ccagcggctg ccgtctttta ttgccagcgc gttacgcaga gggtcctgcc cggaatttgt
     4801 cacttccagc agattattat taagtttgac ggtttttaaa agacgtccat tgtgggtaac
     4861 cagcatggcg ttatcctggg ataaccattt gctttgttct tgttctatat atcccagtac
     4921 cacaaagatt cgttgccctt cattaaggcg taaatacatc gtggaataag gcaacgcctg
     4981 gatctgctga tccgctacgg tgatatcctg attatcgaat aggctggcgc gaaatgtatc
     5041 gaccatactt tgctggctgt gcgtacatcc cgcaagaaat agtgcgaaaa tcgataaaat
     5101 aagagggcgc atcccgggaa ttctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5161 acacattata cgagccgatg attaattgtc aacagctcat ttcagaatat ttgccagaac
     5221 cgttatgatg tcggcgcaaa aaacattatc cagaacggga gtgcgccttg agcgacacga
     5281 attatgcagt gatttacgac ctgcacagcc ataccacagc ttccgatggc tgcctgacgc
     5341 cagaagcatt ggtgcaccgt gcagtcgata agcccggatc cggctgcctc gcgcgtttcg
     5401 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     5461 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     5521 ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc
     5581 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     5641 cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5701 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5761 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5821 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5881 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5941 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6001 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6061 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6121 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6181 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6241 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     6301 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6361 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6421 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6481 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6541 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6601 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6661 ttcatccata gttgcctgac tccccgccgt gtagataact acgatacggg agggcttacc
     6721 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6781 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6841 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6901 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6961 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7021 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7081 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7141 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7201 accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt
     7261 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7321 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7381 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7441 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7501 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7561 aaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttaatttgat
     7621 gcctggcagt ttatggcggg cgtc
//
