LOCUS       sequence-275692-        3329 bp    DNA     circular SYN 15-APR-2025
DEFINITION  expression clone from T7 promoter for full-length (signal peptide 
            containing) B. burgdorferi RevA with C-terminal 
            TEV-protease-cleavable His12.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3329)
  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 3329)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pRevA-TEV-His12
FEATURES             Location/Qualifiers
     source          1..3329
                     /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             580..600
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             601..630
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             604..633
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             607..636
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(730..748)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      744..791
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      888..1343
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(975..994)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1185..1206
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1369..1473
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1474..1542,1543..2334)
                     /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(1692..1711)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2505..3093
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2994..3013
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3247..3264
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcgtaacaa aaacatcttt
      121 aagctgttct ttgcggcgat gctgttcgtg atggcgtgca aagcgtacgt tgaggagaag
      181 aaggagatcg atagcctgat ggaagacgtg ctggcgctgg ttaacgacag cagcggcggt
      241 aaattcaagg attacaagga caagatcaac gagctgaaag aaaacctgaa ggacatcggt
      301 aacgcggagc tgaaagaaaa gctgctgaac ctgcagaaca gcttccaaga caaactggcg
      361 gcgaagctgg cggcgctgaa ggcggcgaaa aacaccattg agaacatcac cgacaaggat
      421 caagacatta gcaagcgtaa gatctggagc gaagcgaaac tggttggtgt gaccgttccg
      481 ctgctgggca gcaacaccag cggtaacggc gataagatga gcaaaaacgc ggtggagcag
      541 attgacaagg ttatcaaatt cctggaggaa ggcaccaacg aaaatcttta ttttcaaggt
      601 caccatcatc accaccatca ccatcatcac caccattaat aaaagggcga attccagcac
      661 actggcggcc gttactagtg gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
      721 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga
      781 ggggtttttt gctgaaagga ggaactatat ccggatctgg cgtaatagcg aagaggcccg
      841 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
      901 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
      961 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     1021 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     1081 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     1141 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     1201 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     1261 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     1321 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
     1381 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     1441 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     1501 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     1561 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     1621 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     1681 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     1741 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     1801 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     1861 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     1921 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     1981 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     2041 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     2101 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     2161 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     2221 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     2281 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     2341 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     2401 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     2461 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     2521 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     2581 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     2641 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     2701 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     2761 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     2821 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     2881 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     2941 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     3001 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     3061 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     3121 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     3181 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     3241 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     3301 cccgcgcgtt ggccgattca ttaatgcag
//
