LOCUS       sequence-275697-        3281 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  empty vector; C-terminal TEV-protease-cleavable His12; T7 promoter; 
            BseRI cloning sites (e.g., In-Fusion compatible).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3281)
  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 3281)
  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: pRSET-TEV-12His
FEATURES             Location/Qualifiers
     source          1..3281
                     /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             532..552
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             553..582
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             556..585
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             559..588
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(682..700)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      696..743
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      840..1295
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(927..946)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1137..1158
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1321..1425
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1426..1494,1495..2286)
                     /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(1644..1663)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2457..3045
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2946..2965
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3199..3216
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata gatctgtcgc tcctcatgta
      121 gacaccgaca ctgtgaacac cgccatcctg ggcgtttcaa ggtccggcaa aggcgagaca
      181 atcgtagtgc caatgatcga caatctgagc cgcgcaaaga accagtctag catggtcgtg
      241 aacgacccca agggcgagct ctactctgcc agcaaggaaa cacttgagaa acgcggatac
      301 gacgtgcagg tgctgaacat cctggatccc ctgcagggca tgagttacaa cccactccag
      361 ctcgtgatcg acgcctgggt gaacggcgac gaccaggagg ctgcaaagag ggcaaacaca
      421 ctgactttct cactgtacaa caacccaaat gctggcgaca acgctttctt taacaccagc
      481 gcccagaatg ctatcaacgg aatcatcctc gctatgagga ggcagaagct tgaaaatctt
      541 tattttcaag gtcaccatca tcaccaccat caccatcatc accaccatta ataaaagggc
      601 gaattccagc acactggcgg ccgttactag tggatccggc tgctaacaaa gcccgaaagg
      661 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
      721 aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggatct ggcgtaatag
      781 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
      841 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
      901 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
      961 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     1021 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     1081 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     1141 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     1201 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     1261 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     1321 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     1381 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     1441 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     1501 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     1561 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     1621 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     1681 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     1741 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     1801 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     1861 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     1921 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     1981 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     2041 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     2101 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     2161 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     2221 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     2281 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     2341 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     2401 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     2461 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     2521 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     2581 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     2641 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     2701 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     2761 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     2821 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     2881 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     2941 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3001 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3061 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     3121 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     3181 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     3241 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca g
//
