LOCUS       sequence-275688-        4091 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression clone from T7 promoter for full-length (signal peptide 
            containing) B. burgdorferi BBA57 with C-terminal 
            TEV-protease-cleavable His12.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4091)
  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 4091)
  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: pBBA57-TEV-His12
FEATURES             Location/Qualifiers
     source          1..4091
                     /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             1342..1362
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1363..1392
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1366..1395
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1369..1398
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1492..1510)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1506..1553
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1650..2105
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1737..1756)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1947..1968
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2131..2235
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2236..2304,2305..3096)
                     /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(2454..2473)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3267..3855
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3756..3775
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4009..4026
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgaacggcaa actgcgtaag
      121 gcgctgaaaa tcgcgatttt caccaccctg ctgctggtga tcagctgcaa cgcgaacatg
      181 gataccaacg acaaaaacaa ggcgctgaac gagtataaac tgaagaacat tagcgaagtt
      241 atcaagaaca gcctgcagct ggagagcgac ccgaaactga agaaagagcc ggaaagcaac
      301 atcaaccaaa gcaccccgcc gatcctggag attgaaaaaa tcgagccggg taaacaggaa
      361 atgagcctga agagcgagtt cggcagcgaa agcctgatgc cgctggaaga gccggaagag
      421 gcgaacatgg cgaaaagcga agaggaaatt gcgaaaatcc aggagaagct gctgctgatt
      481 ggtgcgagcg atgaaatcac cgaccaggag ctgggcgaaa acatgcaaaa gtttctgaac
      541 ccgaccaccg ttgagttcaa aatcagcacc accaccaacg aaaccattct gaccaccatc
      601 gaggaagagg aaatcaacaa caacaaggat aagatcttcg accacaaaga ggagaacgtg
      661 accctgggta acaacagcct ggagaacgcg accctgaaca agaacaccat taccctggcg
      721 cagaaccaaa aatataccac ccacctgaaa aacgacgata agtttatcac caaagaatac
      781 ctgaagcaag ttcgtgatag cctggacaag gcgctgaacg cgattaaaaa cctggagacc
      841 agcaaggagt tccgtgagct ggaaaacctg ccgaaagatc aggagaacag cccgagcgcg
      901 aagaaagaga gcaccgaaag caacattaac aacaccaaca ccgagaacat cgaaatcatt
      961 aaaaaccgtc tgctggagga actgaacaag agcgagctga tcttcgaaga taccaaagac
     1021 ccgctgggca ccagcaccat taagaaagtg atcgacgcgg cgaagaaatg gcaggagaag
     1081 gaaaacagca gccaaatcga ctgggatctg ggcttcaaat ttcacccgaa cccgaaattt
     1141 tataacaagc tgaccgcgca agagtacaaa gtgctggcgg aaaagttcac caaagttaag
     1201 aacgagtaca agaacaccaa agagcagctg aaagcggaaa gcaagctgac caccaacagc
     1261 atcagcaaaa tcattaacgc gaccaaggag ttcgcgaacc aagtgattaa cctgatcctg
     1321 ctggttgaga agaactacca ggaaaatctt tattttcaag gtcaccatca tcaccaccat
     1381 caccatcatc accaccatta ataaaagggc gaattccagc acactggcgg ccgttactag
     1441 tggatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
     1501 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
     1561 gaggaactat atccggatct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     1621 acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
     1681 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     1741 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     1801 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     1861 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     1921 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     1981 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     2041 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     2101 aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     2161 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     2221 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     2281 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     2341 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     2401 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     2461 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     2521 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     2581 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     2641 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     2701 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     2761 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     2821 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     2881 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     2941 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     3001 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     3061 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     3121 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     3181 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     3241 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3301 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3361 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     3421 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     3481 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     3541 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     3601 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     3661 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     3721 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     3781 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     3841 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3901 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3961 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     4021 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     4081 cattaatgca g
//
