LOCUS       sequence-109340-        4520 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses aa1594–1670 of VWF A2 domain flanked by Venus and Cerulean
            and tagged with 7X His at C terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4520)
  AUTHORS   Dayananda KM, Gogia S, Neelamegham S
  TITLE     Escherichia coli-derived von Willebrand factor-A2 domain 
            fluorescence/Forster resonance energy transfer proteins that 
            quantify ADAMTS13 activity.
  JOURNAL   Anal Biochem. 2011 Mar 15;410(2):206-13. doi: 
            10.1016/j.ab.2010.12.005. Epub 2010 Dec 10.
  PUBMED    21146487
REFERENCE   2  (bases 1 to 4520)
  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"; doi: 
            "10.1016/j.ab.2010.12.005"; journalName: "Anal Biochem"; date: 
            "2011-03-15- 15"; volume: "410"; issue: "2"; pages: "206-13"
COMMENT     Sequence Label: pRSET-XS-VWF
FEATURES             Location/Qualifiers
     source          1..4520
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     19..38
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        19..37
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             67..89
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(97..99,100..102,103..615)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIE"
     CDS             join(1048..1050,1051..1053,1054..1764)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             1768..1788
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1789..1809
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     primer_bind     complement(1866..1884)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1880..1927
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2024..2479
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2111..2130)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2321..2342
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2560..2664
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2665..2733,2734..3525)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCDTLLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLID*MEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3696..4284
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4185..4204
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4438..4455
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atctcgatcc cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta
       61 gataattttg tttaacttta agaaggagat atacatatgg tgagcaaggg cgaggagctg
      121 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
      181 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagctgatc
      241 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gggctacggc
      301 ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
      361 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
      421 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
      481 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
      541 cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc
      601 cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc
      661 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca gtccaagctg
      721 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
      781 gggatcactc tcggcatgga cgagaccggt cagggtgacc gggagcaggc gcccaacctg
      841 gtctacatgg tcaccggaaa tcctgcctct gatgagatca agaggctgcc tggagacatc
      901 caggtggtgc ccattggagt gggccctaat gccaacgtgc aggagctgga gaggattggc
      961 tggcccaatg cccctatcct catccaggac tttgagacgc tcccccgaga ggctcctgac
     1021 ctggtgctgc agaggtgctg cgttaacatg gtgagcaagg gcgaggagct gttcaccggg
     1081 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     1141 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     1201 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctgggg cgtgcagtgc
     1261 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     1321 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     1381 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     1441 aaggaggacg gcaacatcct ggggcacaag ctggagtaca acgccatcag cgacaacgtc
     1501 tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac
     1561 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     1621 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccaagct gagcaaagac
     1681 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     1741 ctcggcatgg acgagctgta caagttcgaa aacctgtact tccagggcca tcaccatcac
     1801 catcaccatt aagcttgatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     1861 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     1921 ttttttgctg aaaggaggaa ctatatccgg atctggcgta atagcgaaga ggcccgcacc
     1981 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc
     2041 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     2101 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     2161 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagagcttt acggcacctc
     2221 gaccgcaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     2281 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     2341 ggaacaacac tcaaccctat cgcggtctat tcttttgatt tataagggat tttgccgatt
     2401 tcggcctatt ggttaaaaaa tgagctgatt taacaaatat ttaacgcgaa ttttaacaaa
     2461 atattaacgt ttacaatttc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     2521 ttcacaccgc atacaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     2581 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     2641 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     2701 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     2761 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     2821 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     2881 ctgctatgtg atacactatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     2941 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     3001 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     3061 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     3121 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     3181 aacgacgaga gtgacaccac gatgcctgta gcaatgccaa caacgttgcg caaactatta
     3241 actggcgaac tacttactct agcttcccgg caacaattaa tagactgaat ggaggcggat
     3301 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     3361 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     3421 cgctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     3481 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     3541 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     3601 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     3661 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3721 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3781 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3841 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3901 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     3961 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4021 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4081 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4141 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4201 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4261 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctgggc
     4321 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     4381 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     4441 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     4501 tggccgattc attaatgcag
//
