LOCUS       sequence-275690-        3386 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression clone from T7 promoter for full-length (signal peptide 
            containing) B. burgdorferi P13 with C-terminal 
            TEV-protease-cleavable His12.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3386)
  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 3386)
  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: pP13-TEV-His12
FEATURES             Location/Qualifiers
     source          1..3386
                     /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             637..657
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             658..687
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             661..690
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             664..693
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(787..805)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      801..848
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      945..1400
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1032..1051)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1242..1263
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1426..1530
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1531..1599,1600..2391)
                     /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(1749..1768)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2562..3150
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3051..3070
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3304..3321
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgaacaagct gctgatcttc
      121 gtgctggcga ccttttgcgt tttcagcagc tttgcgcagg cgaacgacag caaaaacggc
      181 gcgtttggta tgagcgcggg tgaaaagctg ctggtgtacg agaccagcaa acaggacccg
      241 atcgttccgt tcctgctgaa cctgttcctg ggttttggca ttggtagctt tgcgcaaggc
      301 gacatcctgg gtggcagcct gattctgggt ttcgatgcgg tgggcatcgg tctgattctg
      361 gcgggcgcgt acctggacat caaggcgctg gatggtatta ccaaaaaggc ggcgtttcag
      421 tggacctggg gcaaaggtgt gatgctggcg ggtgtggtta cgatggcggt tacccgtctg
      481 accgagatta tcctgccgtt cacctttgcg aacagctata accgtaaact gaagaacagc
      541 ctgaacgtgg cgctgggtgg cttcgaaccg agctttgacg ttgcgatggg tcagagcagc
      601 gcgctgggct tcgagctgag ctttaaaaag agctacgaaa atctttattt tcaaggtcac
      661 catcatcacc accatcacca tcatcaccac cattaataaa agggcgaatt ccagcacact
      721 ggcggccgtt actagtggat ccggctgcta acaaagcccg aaaggaagct gagttggctg
      781 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
      841 gttttttgct gaaaggagga actatatccg gatctggcgt aatagcgaag aggcccgcac
      901 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
      961 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     1021 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     1081 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     1141 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     1201 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     1261 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     1321 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     1381 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     1441 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     1501 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     1561 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     1621 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     1681 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     1741 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     1801 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     1861 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     1921 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     1981 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     2041 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     2101 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     2161 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     2221 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     2281 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     2341 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     2401 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     2461 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     2521 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2581 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2641 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2701 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2761 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2821 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2881 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     2941 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3001 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3061 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3121 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3181 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     3241 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     3301 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     3361 gcgcgttggc cgattcatta atgcag
//
