LOCUS       sequence-291948-        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"
     RBS             94..116
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(147..166)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(148..166)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(251..268)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(422..1010)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(502..521)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1181..1972,1973..2041))
                     /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     1804..1823
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2042..2146)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2172..2627)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2309..2330)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2521..2540
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(2724..2771)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2767..2785
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(2879..2908)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             complement(2882..2911)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             complement(2885..2914)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             complement(2915..2935)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 tcacggctct ccggcggggt cttgcacgca atgaagctga tcgcaaccac aatcagcagg
       61 ctgatcagct tgtttttgat gttcatatgt atatctcctt cttaaagtta aacaaaatta
      121 tttctagagg gaaaccgttg tggtctccct atagtgagtc gtattaattt cgcgggatcg
      181 agatcctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
      241 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
      301 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      361 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      421 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      481 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      541 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      601 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      661 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      721 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      781 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      841 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      901 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      961 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     1021 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     1081 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     1141 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     1201 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1261 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1321 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1381 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1441 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1501 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1561 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1621 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1681 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1741 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1801 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1861 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1921 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1981 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     2041 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     2101 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     2161 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     2221 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2281 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2341 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2401 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2461 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2521 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2581 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     2641 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagat
     2701 ccggatatag ttcctccttt cagcaaaaaa cccctcaaga cccgtttaga ggccccaagg
     2761 ggttatgcta gttattgctc agcggtggca gcagccaact cagcttcctt tcgggctttg
     2821 ttagcagccg gatccactag taacggccgc cagtgtgctg gaattcgccc ttttattaat
     2881 ggtggtgatg atggtgatgg tggtgatgat ggtgaccttg aaaataaaga ttttctttcg
     2941 ccgggataat gatcttccag ttgctgtgaa tcagatccgg gttttggatt ttctgacggt
     3001 tcgcgaacca aatcttcggc cacagatacg ggtcgttgta cagctttttg ctgatacccc
     3061 acagggtgtt gccaatcttg ataacgtaca gctcgctgct cgcgttgctc tgatacgctt
     3121 ccaggtaacg cgcgctctcc agaaacagtt cgttcgccag acggaagttc ttaacgtttt
     3181 tcgcctccac acctttttcc cacagggtac ggctacgctc aatcagttcc agggtcttaa
     3241 attgctcttg cggcttgctg tttttcgcga tttcaacttt ttcctcatac gccagcacaa
     3301 tcgggatgct aatttccgcc tcgcccagca ggtaggtgtc cttgttggtg ttcagcaggt
     3361 tcaggtggct gttacgttct ttgatgaacg cacggccgtt ccacgggctc ggcttaatca
     3421 gtttgttgtt gctatagatc ggcaggttgc tcgccgcctc cagcgccttc agctgtttgt
     3481 acatacgctc atcggtttcc ttcagcgctt tcgcttcctt cgcgtttttc gccgcttgct
     3541 gcgcacggtt aaacgctttg ctgtacatgt ccagcgcgtt atccaggtca tagttcttgt
     3601 atttacgggt cgcctcgaaa tacaggttgt taacctcatc aatctccagc ggaatccaga
     3661 tgtacgcctc gttcgcttcc gcgtcgttca gatacttctc gatgttttct ttcaggtagt
     3721 tggtcttctc cttcttctca cgggtctcac gaatgatggt cttgtaacgc tccagcactt
     3781 tcagcgcaat ttcgttaccc tcgatcgcct ttttcttgct gaagctctgc ttcatcgctt
     3841 cctccagacg ttccgcctcg ttaaattctt tgctatagaa caggtggcca cgctcacgaa
     3901 tcagttcgtt cttaatatct ttgatgtcac gcagttgaaa gtttttgttg tccggctgcg
     3961 cgatcttcgc gtttttgctt
//
