LOCUS       sequence-416486-        4586 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BiFC assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4586)
  AUTHORS   Huang C, Bernard D, Zhu J, Dash RC, Chu A, Knupp A, Hakey A, Hadden 
            MK, Garmendia A, Tang Y
  TITLE     Small molecules block the interaction between porcine reproductive 
            and respiratory syndrome virus and CD163 receptor and the infection 
            of pig cells.
  JOURNAL   Virol J. 2020 Jul 30;17(1):116. doi: 10.1186/s12985-020-01361-7.
  PUBMED    32727587
REFERENCE   2  (bases 1 to 4586)
  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: "Virol J."; 
            date: "2020-07-30"; volume: "17(1):116. doi"; pages: " 
            10.1186/s12985-020-01361-7"
COMMENT     Sequence Label: pBiFC-VN155(I152L)-SRCR5
FEATURES             Location/Qualifiers
     source          1..4586
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(418..420,421..423,424..882)
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYLTA"
     primer_bind     complement(463..484)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(724..743)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    897..1031
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(921..940)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     975..994
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1142..1158)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1142..1158)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1155..1177)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1166..1182
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1190..1196,1197..1214,1215..1220))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1235..1256
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1373..1390)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1544..2132)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1624..1643)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2303..3094,3095..3163))
                     /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     2926..2945
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3164..3268)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3336..3354
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3392..3414)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3514..3533
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3727..3749
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3741..3758
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3742..3758
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        3785..4088
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4089..4292
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4243..4263
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4289..4313
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          4430..4526
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
ORIGIN
        1 atggcatcaa tgcagaagct gatctcagag gaggacctgc ttatggccat ggaggcccga
       61 attcggccca ggctggttgg aggggacatt ccctgctctg gtcgtgttga agtacaacat
      121 ggagacacgt ggggcaccgt ctgtgattct gacttctctc tggaggcggc cagcgtgctg
      181 tgcagggaac tacagtgcgg cactgtggtt tccctcctgg ggggagctca ctttggagaa
      241 ggaagtggac agatctgggc tgaagaattc cagtgtgagg ggcacgagtc ccacctttca
      301 ctctgcccag tagcaccccg ccctgacggg acatgtagcc acagcaggga cgtcggcgta
      361 gtctgctcag agatctctcg aggtaccgga ggtggcggga gcggaggtgg cgggagtatg
      421 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
      481 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
      541 aagctgaccc tgaagctgat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
      601 gtgaccaccc tgggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag
      661 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
      721 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
      781 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
      841 ctggagtaca actacaacag ccacaacgtc tatctcaccg cctgagcggc cgcggggatc
      901 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
      961 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     1021 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     1081 gggaggtttt ttcggatcct ctagagtcga tctgcaggca tgctagcttg gcgtaatcat
     1141 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     1201 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     1261 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     1321 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     1381 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     1441 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     1501 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     1561 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     1621 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     1681 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     1741 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     1801 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     1861 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     1921 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     1981 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     2041 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     2101 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     2161 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     2221 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     2281 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     2341 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     2401 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     2461 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     2521 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     2581 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     2641 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     2701 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     2761 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     2821 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     2881 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     2941 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     3001 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     3061 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     3121 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     3181 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     3241 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     3301 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     3361 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     3421 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     3481 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     3541 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     3601 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     3661 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     3721 ggttttccca gtcacgacgt tgtaaaacga cggccagtga gttcgagctt gcatgcctgc
     3781 aggtcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     3841 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     3901 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     3961 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     4021 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     4081 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     4141 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     4201 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     4261 gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga
     4321 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
     4381 ctctagagga tccggtacta gaggaactga aaaaccagaa agttaactgg taagtttagt
     4441 ctttttgtct tttatttcag gtcccggatc cggtggtggt gcaaatcaaa gaactgctcc
     4501 tcagtggatg ttgcctttac ttctaggcct gtacggaagt gttacttctg ctctaaaagc
     4561 tgcggaattg tacccgcggg cccacc
//
