LOCUS       sequence-345145-        7399 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes pelC signal sequence followed by His10-GfcD. Should secrete 
            to periplasm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7399)
  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 7399)
  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-10xHis)
FEATURES             Location/Qualifiers
     source          1..7399
                     /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..153
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             178..198
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2305..2322
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2375..2393)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2389..2436
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2560..2579)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2746..2849
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2850..2918,2919..3710)
                     /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(3068..3087)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3881..4469
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4370..4389
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4623..4640
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4655..4794
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4720..4739)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4880..4902
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4896..5087)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5896..6978)
                     /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     6940..6959
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6979..7056)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7262..7281
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     7369..7388
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7369..7387
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    join(7388..7399,1..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 atcaccacca ccaccatcat cattcttctg gtgtagatct gggtaccgag
      181 aacctgtact tccaatccaa tgccgaagta ttaacctacc cggatccgct ggggtcgtcg
      241 caatcagact ttggcggcac aggattgttg cagatgccaa atgcgcgcat cgcaccggaa
      301 ggtgaattca gcgtcaacta ccgggataac gatcaatacc gtttctactc cacgtccgtg
      361 gcgctgttcc catggctgga aggcaccatt cgttatacgg atgtgcgcac acgcaaatat
      421 agccagtggg aagatttcag cggcgatcag tcatacaaag acaaatcttt cgattttaaa
      481 cttcgcctgt gggaagaagg ttacttgcta ccgcaagtgg cgtttggtaa acgtgatatt
      541 gctggtacgg gtctgtttga cggtgaatat ttggtggcca gcaagcaagc ggggccattt
      601 gatttcaccc tcgggatggc atggggctac gccggtaatg cgggcaatat taccaacccg
      661 ttttgccggg tgagcgataa atattgtcat cgtgcagagt ctcacgatgc gggcgatatc
      721 agctttagcg atatctttcg tggcccggct tccatctttg gcggcattga gtatcaaacg
      781 ccgtggaatc ccctgcgtct gaaactcgaa tacgacggca acaattacca gaatgatttc
      841 gctggcaaac tgcctcaggc aagccatttc aacgtcggtg cagtttatcg cgctgccagc
      901 tgggcagacc tcaacctcag ttatgaacgc ggtaacacgt tgatgtttgg tttcacttta
      961 cggaccaatt tcaacgatct gcgccctgcc ctgcgcgata cgccaaaacc ggcatatcaa
     1021 cctgcgcctg aatctgaagg attgcagtac accacggtag caaaccaact taccgccctg
     1081 aagtataacg cgggctttga cgcgccagaa attcagctgc gcgataagac gctgtatatg
     1141 tctggtgagc aatacaaata ccgtgactct cgtgaagcgg tcgatcgtgc caaccggatt
     1201 ctggtgaata acctgccgca aggcgttgag aagattagcg tgacgcaaaa gcgcgagcat
     1261 atggcgatgg tgactaccga aaccgacgta gccagcctgc gcaaacagct ggccggtaca
     1321 gcgcctggta aatcagagca actgcaacaa caacgtgttg aagcagaaga tctttctgcc
     1381 tttggtcggg gctaccgtat tcgtgaagat cgctttagct actctttcaa cccaacactt
     1441 tcacagtcgc tgggcggccc ggaagatttc tatatgttcc agctggggct gatgtccagc
     1501 gcccgctact ggtttaccga ccacctgctg cttgatggcg gtattttcac caatatttac
     1561 aataactacg acaagtttaa gtcttcacta ttacccgcgg actctaccct cccccgcgtg
     1621 cgcacgcata tccgtgacta cgttcgcaat gacgtttatc tcaacaactt gcaggcgaac
     1681 tattttgccg acttaggcaa tggtttctat ggccaggtgt atggcggtta tctggaaacg
     1741 atgtacgccg gtgtcggttc cgagctgctt tatcgcccgc tagatgccag ctgggcgctg
     1801 ggtgtggacg ttaactacgt gaagcaacgt gactgggaca acatgatgcg cttcaccgat
     1861 tattccacac ccacaggttt cgtgacggct tactggaacc cgccgacgct caacggcgta
     1921 ctgatgaaac tcagcgttgg gcaatatctg gcaaaagata aaggcgcaac gatcgacgtc
     1981 gccaaacgct ttgacagcgg cgtggcggta ggggtatggg cggcaatcag taacgtatct
     2041 aaagatgact acggcgaagg cggctttagt aaaggttttt atatctcgat tccgttcgac
     2101 ttgatgacca ttggacctaa ccgcaaccgc gcggtggttt cgtggacacc attgacgcgt
     2161 gatggtggac aaatgctgtc acgcaaatac cagctctatc caatgacggc agagcgagaa
     2221 gtaccggttg gacaataata acattggaag tggataacgg atccgaattc gagctccgtc
     2281 gacaagcttg cggccgcact cgagcaccac caccaccacc actgagatcc ggctgctaac
     2341 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     2401 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
     2461 ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2521 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2581 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2641 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     2701 cacattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     2761 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2821 atgcttcaat aatttgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     2881 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     2941 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     3001 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     3061 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     3121 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     3181 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     3241 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     3301 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     3361 taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac
     3421 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     3481 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     3541 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     3601 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     3661 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     3721 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     3781 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     3841 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3901 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3961 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4021 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4081 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4141 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4201 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4261 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4321 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     4381 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     4441 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4501 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     4561 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     4621 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     4681 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc
     4741 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     4801 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     4861 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     4921 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
     4981 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     5041 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     5101 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     5161 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     5221 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     5281 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     5341 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     5401 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     5461 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     5521 ctcaacgaca ggagcacgat catgcgcacc cgtggggccg ccatgccggc gataatggcc
     5581 tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc
     5641 aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc
     5701 tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag
     5761 acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg
     5821 ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca
     5881 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5941 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt
     6001 tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag
     6061 ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt
     6121 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc
     6181 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
     6241 gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg
     6301 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
     6361 agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc
     6421 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
     6481 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
     6541 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
     6601 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
     6661 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
     6721 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
     6781 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
     6841 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
     6901 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
     6961 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc
     7021 cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat
     7081 gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg
     7141 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
     7201 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
     7261 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     7321 cgatgcgtcc ggcgtagagg atcgagatcg atctcgatcc cgcgaaatta atacgactca
     7381 ctatagggga attgtgagc
//
