LOCUS       sequence-341373-        7338 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 7338)
  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 7338)
  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: pMCSG26(gfcD)
FEATURES             Location/Qualifiers
     source          1..7338
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(99..118)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        285..388
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(389..457,458..1249)
                     /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(607..626)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1420..2008
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1909..1928
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2162..2179
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2194..2336
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2262..2281)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2422..2444
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2438..2629)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3438..4520)
                     /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     4482..4501
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4521..4598)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4804..4823
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4911..4930
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4911..4929
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4930..4954
                     /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             4970..4990
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             7114..7131
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             7182..7199
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7252..7270)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7266..7313
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
      301 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
      361 tgcttcaata atttgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      421 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
      481 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
      541 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
      601 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
      661 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
      721 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
      781 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
      841 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
      901 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
      961 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     1021 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     1081 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     1141 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     1201 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1261 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     1321 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1381 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1441 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1501 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     1561 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     1621 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     1681 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     1741 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     1801 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     1861 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     1921 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     1981 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     2041 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     2101 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     2161 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     2221 tctgtgcggt atttcacacc gcatatatgg tgcactctca gtacaatctg ctctgatgcc
     2281 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     2341 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     2401 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     2461 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     2521 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     2581 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     2641 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
     2701 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
     2761 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg
     2821 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga
     2881 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga
     2941 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc
     3001 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg
     3061 tcctcaacga caggagcacg atcatgcgca cccgtggggc cgccatgccg gcgataatgg
     3121 cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt
     3181 gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt
     3241 cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga
     3301 agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg
     3361 ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta
     3421 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     3481 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
     3541 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
     3601 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
     3661 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata
     3721 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
     3781 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
     3841 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
     3901 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
     3961 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
     4021 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
     4081 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
     4141 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
     4201 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
     4261 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
     4321 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
     4381 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
     4441 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
     4501 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
     4561 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt
     4621 atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc
     4681 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
     4741 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
     4801 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
     4861 cacgatgcgt ccggcgtaga ggatcgagat cgatctcgat cccgcgaaat taatacgact
     4921 cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt tgtttaactt
     4981 taagaaggag tctctcccat gaagaagaac tcttatcttt taagctgcct ggccattgcc
     5041 gtctccagtg cctgtcatgc tgaagtatta acctacccgg atccgctggg gtcgtcgcaa
     5101 tcagactttg gcggcacagg attgttgcag atgccaaatg cgcgcatcgc accggaaggt
     5161 gaattcagcg tcaactaccg ggataacgat caataccgtt tctactccac gtccgtggcg
     5221 ctgttcccat ggctggaagg caccattcgt tatacggatg tgcgcacacg caaatatagc
     5281 cagtgggaag atttcagcgg cgatcagtca tacaaagaca aatctttcga ttttaaactt
     5341 cgcctgtggg aagaaggtta cttgctaccg caagtggcgt ttggtaaacg tgatattgct
     5401 ggtacgggtc tgtttgacgg tgaatatttg gtggccagca agcaagcggg gccatttgat
     5461 ttcaccctcg ggatggcatg gggctacgcc ggtaatgcgg gcaatattac caacccgttt
     5521 tgccgggtga gcgataaata ttgtcatcgt gcagagtctc acgatgcggg cgatatcagc
     5581 tttagcgata tctttcgtgg cccggcttcc atctttggcg gcattgagta tcaaacgccg
     5641 tggaatcccc tgcgtctgaa actcgaatac gacggcaaca attaccagaa tgatttcgct
     5701 ggcaaactgc ctcaggcaag ccatttcaac gtcggtgcag tttatcgcgc tgccagctgg
     5761 gcagacctca acctcagtta tgaacgcggt aacacgttga tgtttggttt cactttacgg
     5821 accaatttca acgatctgcg ccctgccctg cgcgatacgc caaaaccggc atatcaacct
     5881 gcgcctgaat ctgaaggatt gcagtacacc acggtagcaa accaacttac cgccctgaag
     5941 tataacgcgg gctttgacgc gccagaaatt cagctgcgcg ataagacgct gtatatgtct
     6001 ggtgagcaat acaaataccg tgactctcgt gaagcggtcg atcgtgccaa ccggattctg
     6061 gtgaataacc tgccgcaagg cgttgagaag attagcgtga cgcaaaagcg cgagcatatg
     6121 gcgatggtga ctaccgaaac cgacgtagcc agcctgcgca aacagctggc cggtacagcg
     6181 cctggtaaat cagagcaact gcaacaacaa cgtgttgaag cagaagatct ttctgccttt
     6241 ggtcggggct accgtattcg tgaagatcgc tttagctact ctttcaaccc aacactttca
     6301 cagtcgctgg gcggcccgga agatttctat atgttccagc tggggctgat gtccagcgcc
     6361 cgctactggt ttaccgacca cctgctgctt gatggcggta ttttcaccaa tatttacaat
     6421 aactacgaca agtttaagtc ttcactatta cccgcggact ctaccctccc ccgcgtgcgc
     6481 acgcatatcc gtgactacgt tcgcaatgac gtttatctca acaacttgca ggcgaactat
     6541 tttgccgact taggcaatgg tttctatggc caggtgtatg gcggttatct ggaaacgatg
     6601 tacgccggtg tcggttccga gctgctttat cgcccgctag atgccagctg ggcgctgggt
     6661 gtggacgtta actacgtgaa gcaacgtgac tgggacaaca tgatgcgctt caccgattat
     6721 tccacaccca caggtttcgt gacggcttac tggaacccgc cgacgctcaa cggcgtactg
     6781 atgaaactca gcgttgggca atatctggca aaagataaag gcgcaacgat cgacgtcgcc
     6841 aaacgctttg acagcggcgt ggcggtaggg gtatgggcgg caatcagtaa cgtatctaaa
     6901 gatgactacg gcgaaggcgg ctttagtaaa ggtttttata tctcgattcc gttcgacttg
     6961 atgaccattg gacctaaccg caaccgcgcg gtggtttcgt ggacaccatt gacgcgtgat
     7021 ggtggacaaa tgctgtcacg caaataccag ctctatccaa tgacggcaga gcgagaagta
     7081 ccggttggac aaagtgccca ccatcaccat gggcaccacc atcatcatca ttaacggatc
     7141 cgaattcgag ctccgtcgac aagcttgcgg ccgcactcga gcaccaccac caccaccact
     7201 gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
     7261 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
     7321 gaggaactat atccggat
//
