LOCUS       sequence-275693-        3458 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression clone from T7 promoter for full-length (signal peptide 
            containing) B. burgdorferi BB0405 with C-terminal 
            TEV-protease-cleavable His12.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3458)
  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 3458)
  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: pBB0405-TEV-His12
FEATURES             Location/Qualifiers
     source          1..3458
                     /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             709..729
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             730..759
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             733..762
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             736..765
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(859..877)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      873..920
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1017..1472
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1104..1123)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1314..1335
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1498..1602
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1603..1671,1672..2463)
                     /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(1821..1840)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2634..3222
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3123..3142
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3376..3393
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcgtatgct gctggcgacc
      121 atcattctga ttctgaccac cggtctgctg gcggcgcaga gcaaaagcaa gagcatgacc
      181 gaggacgatt tcgactttga taaactgctg gcgaaagagg agagcgtgcg tcgtctgttt
      241 ggcatcggtt tcggcgttgg ttacccgctg gcgaacatca ccattagcgt gccgtatgtt
      301 gacattgatc tgggttacgg tggctttgtg ggcctgaagc cgaacaactt cctgccgtat
      361 gtggttatgg gtgttgatct gctgttcaaa gacgaaattc acaagaacac catgatcagc
      421 ggtggcatcg gcattggtgc ggactggagc aaaggcagcc cggagaaaag caacgaaaag
      481 ctggaggaag aggaagagaa cgaggcgcag caagtggcga gcctgcagaa ccgtattggt
      541 gtggttatcc gtctgccgct ggttattgaa tacagctttc tgaaaaacat cgtgattggc
      601 ttcaaggcgg ttgcgaccat cggcaccacc atgctgctgg gcagcccgat gagctttgag
      661 ggtgcgcgtt tcaactttct gggcaccggt ttcatcaaga tctacatcga aaatctttat
      721 tttcaaggtc accatcatca ccaccatcac catcatcacc accattaata aaagggcgaa
      781 ttccagcaca ctggcggccg ttactagtgg atccggctgc taacaaagcc cgaaaggaag
      841 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
      901 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatctggc gtaatagcga
      961 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc
     1021 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     1081 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     1141 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     1201 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     1261 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     1321 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     1381 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     1441 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     1501 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     1561 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     1621 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     1681 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     1741 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     1801 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     1861 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     1921 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     1981 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     2041 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     2101 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     2161 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     2221 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     2281 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     2341 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     2401 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     2461 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     2521 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     2581 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2641 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2701 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2761 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     2821 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2881 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2941 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3001 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3061 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3121 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3181 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3241 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3301 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3361 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     3421 ccgcctctcc ccgcgcgttg gccgattcat taatgcag
//
