LOCUS       sequence-345146-        7387 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes pelC signal sequence followed by His6-GfcD. Should secrete 
            to periplasm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7387)
  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 7387)
  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: pBH31(gfcD)
FEATURES             Location/Qualifiers
     source          1..7387
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             28..50
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     sig_peptide     58..123
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     CDS             124..141
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             166..186
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2293..2310
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2363..2381)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2377..2424
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2548..2567)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2734..2837
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2838..2906,2907..3698)
                     /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(3056..3075)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3869..4457
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4358..4377
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4611..4628
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4643..4782
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4708..4727)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4868..4890
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4884..5075)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5884..6966)
                     /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     6928..6947
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6967..7044)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7250..7269
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     7357..7376
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7357..7375
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    join(7376..7387,1..13)
                     /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)."
ORIGIN
        1 ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga tatacatatg
       61 aaatacctgc tgccgaccgc tgctgctggt ctgctgctcc tcgctgccca gccggcgatg
      121 gcccaccatc atcatcatca ttcttctggt gtagatctgg gtaccgagaa cctgtacttc
      181 caatccaatg ccgaagtatt aacctacccg gatccgctgg ggtcgtcgca atcagacttt
      241 ggcggcacag gattgttgca gatgccaaat gcgcgcatcg caccggaagg tgaattcagc
      301 gtcaactacc gggataacga tcaataccgt ttctactcca cgtccgtggc gctgttccca
      361 tggctggaag gcaccattcg ttatacggat gtgcgcacac gcaaatatag ccagtgggaa
      421 gatttcagcg gcgatcagtc atacaaagac aaatctttcg attttaaact tcgcctgtgg
      481 gaagaaggtt acttgctacc gcaagtggcg tttggtaaac gtgatattgc tggtacgggt
      541 ctgtttgacg gtgaatattt ggtggccagc aagcaagcgg ggccatttga tttcaccctc
      601 gggatggcat ggggctacgc cggtaatgcg ggcaatatta ccaacccgtt ttgccgggtg
      661 agcgataaat attgtcatcg tgcagagtct cacgatgcgg gcgatatcag ctttagcgat
      721 atctttcgtg gcccggcttc catctttggc ggcattgagt atcaaacgcc gtggaatccc
      781 ctgcgtctga aactcgaata cgacggcaac aattaccaga atgatttcgc tggcaaactg
      841 cctcaggcaa gccatttcaa cgtcggtgca gtttatcgcg ctgccagctg ggcagacctc
      901 aacctcagtt atgaacgcgg taacacgttg atgtttggtt tcactttacg gaccaatttc
      961 aacgatctgc gccctgccct gcgcgatacg ccaaaaccgg catatcaacc tgcgcctgaa
     1021 tctgaaggat tgcagtacac cacggtagca aaccaactta ccgccctgaa gtataacgcg
     1081 ggctttgacg cgccagaaat tcagctgcgc gataagacgc tgtatatgtc tggtgagcaa
     1141 tacaaatacc gtgactctcg tgaagcggtc gatcgtgcca accggattct ggtgaataac
     1201 ctgccgcaag gcgttgagaa gattagcgtg acgcaaaagc gcgagcatat ggcgatggtg
     1261 actaccgaaa ccgacgtagc cagcctgcgc aaacagctgg ccggtacagc gcctggtaaa
     1321 tcagagcaac tgcaacaaca acgtgttgaa gcagaagatc tttctgcctt tggtcggggc
     1381 taccgtattc gtgaagatcg ctttagctac tctttcaacc caacactttc acagtcgctg
     1441 ggcggcccgg aagatttcta tatgttccag ctggggctga tgtccagcgc ccgctactgg
     1501 tttaccgacc acctgctgct tgatggcggt attttcacca atatttacaa taactacgac
     1561 aagtttaagt cttcactatt acccgcggac tctaccctcc cccgcgtgcg cacgcatatc
     1621 cgtgactacg ttcgcaatga cgtttatctc aacaacttgc aggcgaacta ttttgccgac
     1681 ttaggcaatg gtttctatgg ccaggtgtat ggcggttatc tggaaacgat gtacgccggt
     1741 gtcggttccg agctgcttta tcgcccgcta gatgccagct gggcgctggg tgtggacgtt
     1801 aactacgtga agcaacgtga ctgggacaac atgatgcgct tcaccgatta ttccacaccc
     1861 acaggtttcg tgacggctta ctggaacccg ccgacgctca acggcgtact gatgaaactc
     1921 agcgttgggc aatatctggc aaaagataaa ggcgcaacga tcgacgtcgc caaacgcttt
     1981 gacagcggcg tggcggtagg ggtatgggcg gcaatcagta acgtatctaa agatgactac
     2041 ggcgaaggcg gctttagtaa aggtttttat atctcgattc cgttcgactt gatgaccatt
     2101 ggacctaacc gcaaccgcgc ggtggtttcg tggacaccat tgacgcgtga tggtggacaa
     2161 atgctgtcac gcaaatacca gctctatcca atgacggcag agcgagaagt accggttgga
     2221 caataataac attggaagtg gataacggat ccgaattcga gctccgtcga caagcttgcg
     2281 gccgcactcg agcaccacca ccaccaccac tgagatccgg ctgctaacaa agcccgaaag
     2341 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
     2401 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatt ggcgaatggg
     2461 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2521 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2581 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2641 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cattaacgct
     2701 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     2761 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     2821 tttgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2881 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2941 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     3001 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     3061 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     3121 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     3181 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     3241 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     3301 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     3361 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     3421 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     3481 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3541 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3601 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3661 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3721 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3781 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3841 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3901 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3961 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     4021 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4081 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4141 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4201 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4261 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4321 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4381 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4441 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4501 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4561 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     4621 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4681 tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4741 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     4801 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     4861 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     4921 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     4981 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     5041 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     5101 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     5161 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     5221 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta
     5281 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag
     5341 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt
     5401 gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt
     5461 gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg
     5521 agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg
     5581 aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat
     5641 accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg
     5701 acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt
     5761 gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     5821 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg
     5881 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5941 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
     6001 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
     6061 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
     6121 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg
     6181 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     6241 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     6301 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     6361 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     6421 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
     6481 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     6541 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     6601 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
     6661 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
     6721 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     6781 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     6841 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     6901 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     6961 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     7021 ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca
     7081 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
     7141 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     7201 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     7261 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     7321 cgtagaggat cgagatcgat ctcgatcccg cgaaattaat acgactcact ataggggaat
     7381 tgtgagc
//
