LOCUS       sequence-109428-        3800 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses aa 1594-1670 of A2 Domain of VWF protein, TEV cleavage 
            site and Venus followed by 7X His tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3800)
  AUTHORS   Gogia S, Lo CY, Neelamegham S
  TITLE     Detection of Plasma Protease Activity Using Microsphere-Cytometry 
            Assays with E. coli Derived Substrates: VWF Proteolysis by ADAMTS13.
  JOURNAL   PLoS One. 2015 May 18;10(5):e0126556. doi: 
            10.1371/journal.pone.0126556. eCollection 2015.
  PUBMED    25992814
REFERENCE   2  (bases 1 to 3800)
  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: "PLoS One.";
            date: "2015-05-18"; volume: "10(5):e0126556. doi"; pages: " 
            10.1371/journal.pone.0126556. eCollection 2015"
COMMENT     Sequence Label: pRSET-A2-77p-Venus
FEATURES             Location/Qualifiers
     source          1..3800
                     /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             340..360
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(361..363,364..366,367..879)
                     /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"
     primer_bind     complement(406..427)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(667..686)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1014..1035
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1069..1089
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     primer_bind     complement(1146..1164)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1160..1207
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1304..1759
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1391..1410)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1601..1622
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1840..1944
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1945..2013,2014..2805)
                     /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      2976..3564
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3465..3484
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3718..3735
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atctcgatcc cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta
       61 gataattttg tttaacttta agaaggagat atacatatga aacagggtga ccgggagcag
      121 gcgcccaacc tggtctacat ggtcaccgga aatcctgcct ctgatgagat caggaggctg
      181 cctggagaca tccaggtggt gcccattgga gtgggcccta atgccaacgt gcaggagctg
      241 gagaggattg gctggcccaa tgcccctatc ctcatccagg actttgagac gctcccccga
      301 gaggctcctg acctggtgct gcagaggtgc tgcttcgaag agaatctata ctttcaaggc
      361 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
      421 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      481 ggcaagctga ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc
      541 ctcgtgacca ccctgggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag
      601 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      661 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      721 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      781 aagctggagt acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac
      841 ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc
      901 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      961 tacctgagct accagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     1021 ctgctggagt tcgtgaccgc cgccgggatc actctaggca tggacgagca tcatcaccac
     1081 caccatcact aagcttgatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     1141 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     1201 ttttttgctg aaaggaggaa ctatatccgg atctggcgta atagcgaaga ggcccgcacc
     1261 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc
     1321 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     1381 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     1441 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagagcttt acggcacctc
     1501 gaccgcaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     1561 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     1621 ggaacaacac tcaaccctat cgcggtctat tcttttgatt tataagggat tttgccgatt
     1681 tcggcctatt ggttaaaaaa tgagctgatt taacaaatat ttaacgcgaa ttttaacaaa
     1741 atattaacgt ttacaatttc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     1801 ttcacaccgc atacaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     1861 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     1921 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     1981 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     2041 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     2101 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     2161 ctgctatgtg atacactatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     2221 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     2281 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     2341 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     2401 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     2461 aacgacgaga gtgacaccac gatgcctgta gcaatgccaa caacgttgcg caaactatta
     2521 actggcgaac tacttactct agcttcccgg caacaattaa tagactgaat ggaggcggat
     2581 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     2641 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     2701 cgctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     2761 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     2821 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     2881 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     2941 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3001 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3061 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3121 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3181 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     3241 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     3301 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     3361 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     3421 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     3481 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     3541 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctgggc
     3601 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     3661 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     3721 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     3781 tggccgattc attaatgcag
//
