LOCUS       sequence-275699-        4271 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression clone from T7 promoter for full-length (signal peptide 
            containing) B. burgdorferi HtrA with C-terminal 
            TEV-protease-cleavable His12.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4271)
  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 4271)
  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: pHtrA-TEV-His12
FEATURES             Location/Qualifiers
     source          1..4271
                     /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             1522..1542
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1543..1572
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1546..1575
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1549..1578
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1672..1690)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1686..1733
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1830..2285
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1917..1936)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2127..2148
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2311..2415
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2416..2484,2485..3276)
                     /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(2634..2653)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3447..4035
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3936..3955
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4189..4206
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tggaaaagaa attctttagc
      121 ggttttctgc tgagcttcct ggcgctgagc attggtttct ttatcggcat gcactatctg
      181 gcgagcaacc gtagcaacat cgtgtttgcg gaggaaaaag ataacaccgt tcgtgcgctg
      241 caagacagct tccgtgaggt gagcaagaaa attctgccga gcagcgttga agtgcacgcg
      301 accggtgtta ttaagcagag ctttccgatc ccgttctttt tctttgatat gccggagttc
      361 gacagcgaac gtaaaagcaa ctgggcgggt agcggcgtta tcattggccg tgacagccag
      421 aagaaaagcc tgttttacgt ggttaccaac agccacgtgg ttgataaggc gaccgagctg
      481 gaagtggtta gctatgacaa gaaaaagcac aaagcgaagc tgatcggtaa agatgagaaa
      541 aaggacattg cgctgatcag cttcgaaagc gacgatgcga ccattaaggt ggcggatctg
      601 ggcgacagcg ataaactgga gatcggtgac tgggtgatgg cggttggcag cccgttccaa
      661 tttagcttca ccgtgaccgc gggtatcgtt agcggtctgc agcgtagcgc gaacccgaac
      721 ctgcagagcc gtaacctgtt cattcaaacc gatgcggcga tcaaccgtgg taacagcggc
      781 ggtccgctgg tgaacatcaa gggtgaggtt atcggcatta acgcgtggat tgcgagcaac
      841 agcggcggta acatcggtct gggctttgcg attccggtga acaacatcaa aagcaccgtt
      901 gacttctttc tgaagggtaa aaagattgaa agcgcgtggc tgggcatcag cttctacccg
      961 ctgaaaaccc gtgatagcga ggtgctgaag agcctgggtg tggaaagcaa cgacgttagc
     1021 gcggcgatca ttgcgagcct gtatccgggc agcccggcgg ttaagagcgg tctgcgtgcg
     1081 ggcgacatca ttatgaaagt gaacggtgtt agcatgagcg tgtttcaaga tgttaccagc
     1141 tatattagcg acttctacgc gggcgagaag gttaacgtgg aaatcctgcg tggcaacgtg
     1201 aaaaagaaca tcgagattgt gctggcggtt cgtccgaaag ataaggagct gagcagcagc
     1261 aagatgctgc cgggttttgt ggtttacccg ctggttgagg acatcaaagc gcagctgaac
     1321 ctgcgtaact ggattaaggg tgtggttgtg gattacatcg acaaaaacct ggcgagcaac
     1381 attaaaatga agagcggcga cgtgatcctg agcgttaaca gcaaaagcgt gagcaacctg
     1441 cgtgagttct atgatgcgct ggaagttggt aaaaacacct acaagatcct gcgtggcaac
     1501 gacagcttta agattacctt cgaaaatctt tattttcaag gtcaccatca tcaccaccat
     1561 caccatcatc accaccatta ataaaagggc gaattccagc acactggcgg ccgttactag
     1621 tggatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
     1681 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
     1741 gaggaactat atccggatct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     1801 acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
     1861 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     1921 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     1981 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     2041 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     2101 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     2161 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     2221 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     2281 aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     2341 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     2401 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     2461 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     2521 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     2581 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     2641 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     2701 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     2761 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     2821 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     2881 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     2941 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     3001 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     3061 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     3121 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     3181 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     3241 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     3301 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     3361 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     3421 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3481 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3541 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     3601 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     3661 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     3721 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     3781 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     3841 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     3901 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     3961 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4021 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     4081 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     4141 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     4201 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     4261 cattaatgca g
//
