LOCUS       sequence-275698-        3875 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression clone from T7 promoter for full-length (signal peptide 
            containing) B. burgdorferi DipA with C-terminal 
            TEV-protease-cleavable His12.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3875)
  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 3875)
  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: pDipA-TEV-His12
FEATURES             Location/Qualifiers
     source          1..3875
                     /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             1126..1146
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1147..1176
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1150..1179
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1153..1182
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1276..1294)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1290..1337
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1434..1889
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1521..1540)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1731..1752
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1915..2019
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2020..2088,2089..2880)
                     /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(2238..2257)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3051..3639
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3540..3559
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3793..3810
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgctgatcaa aaagatttgc
      121 ttctgcctgc tgctgctgca ggcgaagatt atcctgagcc aagatcagca aaacctggaa
      181 tttaacaacg agtacttcta cttcaacttc caaagcgcgt actatccgcc gcacgaactg
      241 ggcaccaacg gcagcggttt cagccagagc tttaaacacc cgaacttcca aaacatcgac
      301 ccgagcctga aaattccggt gaactactgg ggtggcatca agctgattag ctacctgggt
      361 tactacaaga acttcaaggc gctgaacaac ccgaacagca tcttctttaa gaacaacggc
      421 atcgacattg atctgaacat cggtctgagc ccggtgattg ttagcttcaa aagcaccgtt
      481 agcttcaccc cgatcgcgtt tctgaacctg tacgcgagca acgaactggg cattggttgg
      541 caggcgtttg gcttcaaagg cgtgggtgtt cacatcgaga acggcaagta cagcaacgcg
      601 gtggaatttt atagcgagat caccgcgggt ggccgttttc agttcgacct gaacgcgatt
      661 ttccaaggcg agtggaccca cattatcacc gttatcggta acgactttac ctacctgatt
      721 aacccgcacg cgaaaaagga tcaactgtgg aaatataagg cggacgatgg caagaacatc
      781 aacggtatgc tgattaaacc gtacgcgctg ctggcgtata agatgccgat cccgctgaac
      841 accattggcc tgctgtatga aggtaaaacc cagatcggca aggagcgtaa cattagcagc
      901 tggaaaaaca agggctgggg tagcgacatc ttctaccaca acatcagcat tgttgcgaac
      961 tttgagatta tcaaaccgct gaccctgggc ctgcaattca agctgagcac caacccgacc
     1021 tataccagca acaccgcggg tctggcggac atcagcaagc gtattgcgac cggcaacagc
     1081 tacttttact atgatagcat cggtatgagc atcacctaca agtacgaaaa tctttatttt
     1141 caaggtcacc atcatcacca ccatcaccat catcaccacc attaataaaa gggcgaattc
     1201 cagcacactg gcggccgtta ctagtggatc cggctgctaa caaagcccga aaggaagctg
     1261 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
     1321 tcttgagggg ttttttgctg aaaggaggaa ctatatccgg atctggcgta atagcgaaga
     1381 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
     1441 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     1501 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     1561 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     1621 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     1681 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     1741 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     1801 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     1861 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     1921 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     1981 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     2041 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     2101 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     2161 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     2221 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     2281 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2341 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2401 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2461 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2521 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     2581 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2641 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     2701 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     2761 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     2821 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     2881 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     2941 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3001 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3061 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3121 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3181 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     3241 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3301 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3361 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3421 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3481 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3541 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3601 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3661 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3721 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3781 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     3841 cctctccccg cgcgttggcc gattcattaa tgcag
//
