LOCUS       sequence-291947-        3617 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression clone from T7 promoter for full-length (signal peptide 
            containing) B. burgdorferi BB0238 with C-terminal 
            TEV-protease-cleavable His12.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3617)
  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 3617)
  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: pBB0238-TEV-His12
FEATURES             Location/Qualifiers
     source          1..3617
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      310..898
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     799..818
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1052..1069
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1154..1173
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1154..1172
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1204..1226
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2002..2022
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2023..2052
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2026..2055
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2029..2058
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(2152..2170)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2166..2213
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2310..2765
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2397..2416)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2607..2628
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2791..2895
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2896..2964,2965..3617,1..139)
                     /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(3114..3133)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
       61 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
      121 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
      181 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
      241 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
      301 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
      361 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
      421 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
      481 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
      541 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
      601 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
      661 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
      721 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
      781 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
      841 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
      901 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
      961 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     1021 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     1081 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcaggatctc
     1141 gatcccgcga aattaatacg actcactata gggagaccac aacggtttcc ctctagaaat
     1201 aattttgttt aactttaaga aggagatata catatgcgtc gtctgttcct gctgtatatc
     1261 ctgtgcagct ttgttttcct gaacctgttt gcgcaaggta gcagcagcta cattgacaaa
     1321 cagaaggagc tggcgatctt ctactatgaa gtgggccaac gttacatcaa cgttggcaag
     1381 atcaaaaagg gcaagctgtt tcaggcgaag gcgctgaaaa tttacccgga tctgaaaaag
     1441 ggtttcgaca tcaaactggc ggtgaaggag ctggatgcgc gtattaaaga cgataacccg
     1501 aaggttgtga tgctggaaga catcaagctg gaggaaattc cgggcatcgt tcacgagaag
     1561 atcgaaatca acgatttcac caacgcgccg aagattgagt acatcgcgca gcgtgaacgt
     1621 agcaagaacc aagacaagat catcaagttc cagttcggca agttcgcgcg tgcgctgatt
     1681 agccgtaact ttgacctgtt cgacagcgtg atcgcggata aggtgaacgt tatgggccaa
     1741 tttgaaagca aaaacgactt catcagcacc ctgagcagcg cgagcagcaa agcggatgcg
     1801 gacgagctgg aatacctgag cgtggacgat tactatgatc tgaagagcct gaaaatcagc
     1861 aagagcaacg acaccagctt cgcggtgaac gttaacgcga aaaagaacga cgttaccaaa
     1921 aactttccgt tctggaagga gcgtcagacc ctgattttta ccaccgaaga cgataacaac
     1981 tggttcctga gcagcatcaa cgaaaatctt tattttcaag gtcaccatca tcaccaccat
     2041 caccatcatc accaccatta ataaaagggc gaattccagc acactggcgg ccgttactag
     2101 tggatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
     2161 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
     2221 gaggaactat atccggatct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     2281 acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
     2341 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     2401 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     2461 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     2521 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     2581 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     2641 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     2701 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     2761 aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     2821 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     2881 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     2941 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     3001 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     3061 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     3121 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     3181 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     3241 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     3301 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     3361 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     3421 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     3481 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     3541 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     3601 ggtgagcgtg ggtctcg
//
