LOCUS       sequence-345638-        6250 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gfcD protein expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6250)
  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 6250)
  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(gfcD)
FEATURES             Location/Qualifiers
     source          1..6250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(522..539)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      692..778
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      870..897
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        916..1007
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1008..1076,1077..1868)
                     /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(1226..1245)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2039..2627
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2528..2547
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2781..2798
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2813..2953
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2879..2898)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3039..3061
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        join(3286..3291,3292..3307,3308..3314)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    3322..3338
                     /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     3327..3349
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             5455..5472
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        5536..5613
                     /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             join(5614..6250,1..446)
                     /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(5633..5652)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc
       61 tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg
      121 caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat
      181 acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt
      241 ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg
      301 tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca
      361 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
      421 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gcgcgaattg
      481 tcgaagcttg gctgttttgg cggatgagag aagattttca gcctgataca gattaaatca
      541 gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca
      601 cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct
      661 ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga
      721 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
      781 gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc
      841 gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc
      901 gtttctacaa actcttttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
      961 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     1021 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     1081 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     1141 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     1201 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     1261 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     1321 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     1381 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     1441 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     1501 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     1561 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     1621 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     1681 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     1741 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg gcggggagtc
     1801 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     1861 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     1921 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     1981 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2041 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2101 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2161 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     2221 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2281 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2341 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2401 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     2461 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     2521 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     2581 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     2641 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     2701 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     2761 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     2821 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta
     2881 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     2941 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3001 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3061 gttttcaccg tcatcaccga aacgcgcgag gcagccggat ccgggcttat cgactgcacg
     3121 gtgcaccaat gcttctggcg tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc
     3181 gtaaatcact gcataattcg tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt
     3241 gcgccgacat cataacggtt ctggcaaata ttctgaaatg agctgttgac aattaatcat
     3301 cggctcgtat aatgtgtgga attgtgagcg gataacaatt tcacacagga aacagaattg
     3361 atgaagaaga actcttatct tttaagctgc ctggccattg ccgtctccag tgcctgtcat
     3421 gctgaagtat taacctaccc ggatccgctg gggtcgtcgc aatcagactt tggcggcaca
     3481 ggattgttgc agatgccaaa tgcgcgcatc gcaccggaag gtgaattcag cgtcaactac
     3541 cgggataacg atcaataccg tttctactcc acgtccgtgg cgctgttccc atggctggaa
     3601 ggcaccattc gttatacgga tgtgcgcaca cgcaaatata gccagtggga agatttcagc
     3661 ggcgatcagt catacaaaga caaatctttc gattttaaac ttcgcctgtg ggaagaaggt
     3721 tacttgctac cgcaagtggc gtttggtaaa cgtgatattg ctggtacggg tctgtttgac
     3781 ggtgaatatt tggtggccag caagcaagcg gggccatttg atttcaccct cgggatggca
     3841 tggggctacg ccggtaatgc gggcaatatt accaacccgt tttgccgggt gagcgataaa
     3901 tattgtcatc gtgcagagtc tcacgatgcg ggcgatatca gctttagcga tatctttcgt
     3961 ggcccggctt ccatctttgg cggcattgag tatcaaacgc cgtggaatcc cctgcgtctg
     4021 aaactcgaat acgacggcaa caattaccag aatgatttcg ctggcaaact gcctcaggca
     4081 agccatttca acgtcggtgc agtttatcgc gctgccagct gggcagacct caacctcagt
     4141 tatgaacgcg gtaacacgtt gatgtttggt ttcactttac ggaccaattt caacgatctg
     4201 cgccctgccc tgcgcgatac gccaaaaccg gcatatcaac ctgcgcctga atctgaagga
     4261 ttgcagtaca ccacggtagc aaaccaactt accgccctga agtataacgc gggctttgac
     4321 gcgccagaaa ttcagctgcg cgataagacg ctgtatatgt ctggtgagca atacaaatac
     4381 cgtgactctc gtgaagcggt cgatcgtgcc aaccggattc tggtgaataa cctgccgcaa
     4441 ggcgttgaga agattagcgt gacgcaaaag cgcgagcata tggcgatggt gactaccgaa
     4501 accgacgtag ccagcctgcg caaacagctg gccggtacag cgcctggtaa atcagagcaa
     4561 ctgcaacaac aacgtgttga agcagaagat ctttctgcct ttggtcgggg ctaccgtatt
     4621 cgtgaagatc gctttagcta ctctttcaac ccaacacttt cacagtcgct gggcggcccg
     4681 gaagatttct atatgttcca gctggggctg atgtccagcg cccgctactg gtttaccgac
     4741 cacctgctgc ttgatggcgg tattttcacc aatatttaca ataactacga caagtttaag
     4801 tcttcactat tacccgcgga ctctaccctc ccccgcgtgc gcacgcatat ccgtgactac
     4861 gttcgcaatg acgtttatct caacaacttg caggcgaact attttgccga cttaggcaat
     4921 ggtttctatg gccaggtgta tggcggttat ctggaaacga tgtacgccgg tgtcggttcc
     4981 gagctgcttt atcgcccgct agatgccagc tgggcgctgg gtgtggacgt taactacgtg
     5041 aagcaacgtg actgggacaa catgatgcgc ttcaccgatt attccacacc cacaggtttc
     5101 gtgacggctt actggaaccc gccgacgctc aacggcgtac tgatgaaact cagcgttggg
     5161 caatatctgg caaaagataa aggcgcaacg atcgacgtcg ccaaacgctt tgacagcggc
     5221 gtggcggtag gggtatgggc ggcaatcagt aacgtatcta aagatgacta cggcgaaggc
     5281 ggctttagta aaggttttta tatctcgatt ccgttcgact tgatgaccat tggacctaac
     5341 cgcaaccgcg cggtggtttc gtggacacca ttgacgcgtg atggtggaca aatgctgtca
     5401 cgcaaatacc agctctatcc aatgacggca gagcgagaag taccggttgg acaacatcac
     5461 catcaccatc actaactgca ggcagccaag cttcgacaat tcgcgcgcga aggcgaagcg
     5521 gcatgcattt acgttgacac catcgaatgg tgcaaaacct ttcgcggtat ggcatgatag
     5581 cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc
     5641 gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac
     5701 gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc
     5761 aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc
     5821 agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa
     5881 ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg
     5941 gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat
     6001 gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat
     6061 gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg
     6121 ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta
     6181 agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa
     6241 attcagccga
//
