LOCUS       sequence-345637-        6262 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 6262)
  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 6262)
  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-10xHis)
FEATURES             Location/Qualifiers
     source          1..6262
                     /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"
     CDS             4971..5000
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     promoter        5064..5141
                     /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             5142..6224
                     /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(5161..5180)
                     /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 gaattgatga
     2881 agaagaactc ttatctttta agctgcctgg ccattgccgt ctccagtgcc tgtcatgctg
     2941 aagtattaac ctacccggat ccgctggggt cgtcgcaatc agactttggc ggcacaggat
     3001 tgttgcagat gccaaatgcg cgcatcgcac cggaaggtga attcagcgtc aactaccggg
     3061 ataacgatca ataccgtttc tactccacgt ccgtggcgct gttcccatgg ctggaaggca
     3121 ccattcgtta tacggatgtg cgcacacgca aatatagcca gtgggaagat ttcagcggcg
     3181 atcagtcata caaagacaaa tctttcgatt ttaaacttcg cctgtgggaa gaaggttact
     3241 tgctaccgca agtggcgttt ggtaaacgtg atattgctgg tacgggtctg tttgacggtg
     3301 aatatttggt ggccagcaag caagcggggc catttgattt caccctcggg atggcatggg
     3361 gctacgccgg taatgcgggc aatattacca acccgttttg ccgggtgagc gataaatatt
     3421 gtcatcgtgc agagtctcac gatgcgggcg atatcagctt tagcgatatc tttcgtggcc
     3481 cggcttccat ctttggcggc attgagtatc aaacgccgtg gaatcccctg cgtctgaaac
     3541 tcgaatacga cggcaacaat taccagaatg atttcgctgg caaactgcct caggcaagcc
     3601 atttcaacgt cggtgcagtt tatcgcgctg ccagctgggc agacctcaac ctcagttatg
     3661 aacgcggtaa cacgttgatg tttggtttca ctttacggac caatttcaac gatctgcgcc
     3721 ctgccctgcg cgatacgcca aaaccggcat atcaacctgc gcctgaatct gaaggattgc
     3781 agtacaccac ggtagcaaac caacttaccg ccctgaagta taacgcgggc tttgacgcgc
     3841 cagaaattca gctgcgcgat aagacgctgt atatgtctgg tgagcaatac aaataccgtg
     3901 actctcgtga agcggtcgat cgtgccaacc ggattctggt gaataacctg ccgcaaggcg
     3961 ttgagaagat tagcgtgacg caaaagcgcg agcatatggc gatggtgact accgaaaccg
     4021 acgtagccag cctgcgcaaa cagctggccg gtacagcgcc tggtaaatca gagcaactgc
     4081 aacaacaacg tgttgaagca gaagatcttt ctgcctttgg tcggggctac cgtattcgtg
     4141 aagatcgctt tagctactct ttcaacccaa cactttcaca gtcgctgggc ggcccggaag
     4201 atttctatat gttccagctg gggctgatgt ccagcgcccg ctactggttt accgaccacc
     4261 tgctgcttga tggcggtatt ttcaccaata tttacaataa ctacgacaag tttaagtctt
     4321 cactattacc cgcggactct accctccccc gcgtgcgcac gcatatccgt gactacgttc
     4381 gcaatgacgt ttatctcaac aacttgcagg cgaactattt tgccgactta ggcaatggtt
     4441 tctatggcca ggtgtatggc ggttatctgg aaacgatgta cgccggtgtc ggttccgagc
     4501 tgctttatcg cccgctagat gccagctggg cgctgggtgt ggacgttaac tacgtgaagc
     4561 aacgtgactg ggacaacatg atgcgcttca ccgattattc cacacccaca ggtttcgtga
     4621 cggcttactg gaacccgccg acgctcaacg gcgtactgat gaaactcagc gttgggcaat
     4681 atctggcaaa agataaaggc gcaacgatcg acgtcgccaa acgctttgac agcggcgtgg
     4741 cggtaggggt atgggcggca atcagtaacg tatctaaaga tgactacggc gaaggcggct
     4801 ttagtaaagg tttttatatc tcgattccgt tcgacttgat gaccattgga cctaaccgca
     4861 accgcgcggt ggtttcgtgg acaccattga cgcgtgatgg tggacaaatg ctgtcacgca
     4921 aataccagct ctatccaatg acggcagagc gagaagtacc ggttggacaa catcaccatc
     4981 accatcatca ccaccatcac taactgcagg cagccaagct tcgacaattc gcgcgcgaag
     5041 gcgaagcggc atgcatttac gttgacacca tcgaatggtg caaaaccttt cgcggtatgg
     5101 catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat
     5161 acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg
     5221 ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt
     5281 acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg
     5341 ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg
     5401 ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct
     5461 gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc
     5521 cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat
     5581 ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat gaagacggta
     5641 cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg
     5701 gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc
     5761 gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc
     5821 aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg
     5881 atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg
     5941 atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa
     6001 ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac
     6061 tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa
     6121 aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     6181 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     6241 gtaagttagc gcgaattgtc ga
//
