LOCUS       sequence-265247-        3980 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression clone from T7 promoter for full-length (signal peptide 
            containing) B. burgdorferi BB0323 with C-terminal 
            TEV-protease-cleavable His12.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3980)
  AUTHORS   Robertson KE, Truong CD, Craciunescu FM, Chiu PL, Fromme P, Hansen 
            DT
  TITLE     Membrane directed expression in Escherichia coli of BBA57 and other 
            virulence factors from the Lyme disease agent Borrelia burgdorferi.
  JOURNAL   Sci Rep. 2019 Nov 26;9(1):17606. doi: 10.1038/s41598-019-53830-x.
  PUBMED    31772280
REFERENCE   2  (bases 1 to 3980)
  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: "Sci Rep."; 
            date: "2019-11-26"; volume: "9(1):17606. doi"; pages: " 
            10.1038/s41598-019-53830-x"
COMMENT     Sequence Label: pBB0323-TEV-His12
FEATURES             Location/Qualifiers
     source          1..3980
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             70..92
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1231..1251
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1252..1281
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1255..1284
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1258..1287
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1381..1399)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1395..1442
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1539..1994
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1626..1645)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1836..1857
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2020..2124
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2125..2193,2194..2985)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2343..2362)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3156..3744
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3645..3664
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3898..3915
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgaacatcaa aaacaagctg
      121 atcagcctgc tgattgtggt tgcgatcagc ttcattgcgt gcaagacccc gccggagagc
      181 cgtgaaagca aaaacgcgaa gatcgcgcag ccggacaaca aaaactttca actgcgtgac
      241 atcaaagata ttaagaacga actgattcgt gagcgtggcc acctgttcta tagcaaagaa
      301 tttaacgagg cggaacgtct ggaggaagcg atgaagcaga gcttcagcaa gaaaaaggcg
      361 atcgagggta acgaaattgc gctgaaagtg ctggagcgtt acaagaccat cattcgtgag
      421 acccgtgaga agaaggagaa gaccaactac ctgaaagaaa acatcgagaa gtatctgaac
      481 gacgcggaag cgaacgaggc gtacatctgg attccgctgg agattgatga ggttaacaac
      541 ctgtatttcg aggcgacccg taaatacaag aactatgacc tggataacgc gctggacatg
      601 tacagcaaag cgtttaaccg tgcgcagcaa gcggcgaaaa acgcgaagga agcgaaagcg
      661 ctgaaggaaa ccgatgagcg tatgtacaaa cagctgaagg cgctggaggc ggcgagcaac
      721 ctgccgatct atagcaacaa caaactgatt aagccgagcc cgtggaacgg ccgtgcgttc
      781 atcaaagaac gtaacagcca cctgaacctg ctgaacacca acaaggacac ctacctgctg
      841 ggcgaggcgg aaattagcat cccgattgtg ctggcgtatg aggaaaaagt tgaaatcgcg
      901 aaaaacagca agccgcaaga gcaatttaag accctggaac tgattgagcg tagccgtacc
      961 ctgtgggaaa aaggtgtgga ggcgaaaaac gttaagaact tccgtctggc gaacgaactg
     1021 tttctggaga gcgcgcgtta cctggaagcg tatcagagca acgcgagcag cgagctgtac
     1081 gttatcaaga ttggcaacac cctgtggggt atcagcaaaa agctgtacaa cgacccgtat
     1141 ctgtggccga agatttggtt cgcgaaccgt cagaaaatcc aaaacccgga tctgattcac
     1201 agcaactgga agatcattat cccggcgaaa gaaaatcttt attttcaagg tcaccatcat
     1261 caccaccatc accatcatca ccaccattaa taaaagggcg aattccagca cactggcggc
     1321 cgttactagt ggatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
     1381 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     1441 tgctgaaagg aggaactata tccggatctg gcgtaatagc gaagaggccc gcaccgatcg
     1501 cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt
     1561 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     1621 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     1681 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     1741 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     1801 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     1861 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     1921 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     1981 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     2041 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     2101 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     2161 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     2221 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     2281 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     2341 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     2401 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     2461 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     2521 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     2581 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     2641 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     2701 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     2761 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     2821 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     2881 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     2941 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     3001 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     3061 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     3121 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3181 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3241 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3301 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3361 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     3421 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     3481 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     3541 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     3601 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     3661 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     3721 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     3781 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     3841 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     3901 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     3961 tggccgattc attaatgcag
//
