LOCUS Exported 4088 bp ds-DNA circular SYN 14-FEB-2022 DEFINITION Mammalian expression of a fluorescent protein fragment derived from Venus for BiFC and multicolor BiFC assays. ACCESSION . VERSION . KEYWORDS pBiFC-VC155 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4088) AUTHORS Shyu YJ, Liu H, Deng X, Hu CD TITLE Identification of new fluorescent protein fragments for bimolecular fluorescence complementation analysis under physiological conditions. JOURNAL Biotechniques. 2006 Jan . 40(1):61-6. PUBMED 16454041 REFERENCE 2 (bases 1 to 4088) AUTHORS . TITLE Direct Submission JOURNAL Exported Feb 14, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4088 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 1..17 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" enhancer 44..347 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 348..551 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 502..522 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 548..572 /label=LNCX /note="Human CMV promoter, forward primer" intron 689..785 /label=SV40 intron /note="modified SV40 intron with splice donor and acceptor sites" CDS 849..875 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 978..1229 /codon_start=1 /product="C-terminal fragment of mVenus for use in bimolecular fluorescence complementation (BiFC) (Kodama and Hu, 2010)" /label=VC155 /translation="DKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH YLSYQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK" primer_bind 1166..1187 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 1244..1378 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1268..1287) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1322..1341 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(1489..1505) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1489..1505) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1502..1524) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1513..1529 /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(1537..1567) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1582..1603 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1720..1737) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1891..2479) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1971..1990) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2650..3510) /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 3273..3292 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3511..3615) /gene="bla" /label=AmpR promoter primer_bind 3683..3701 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(3739..3761) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 3861..3880 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4074..8 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 4088..17 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" ORIGIN 1 gtaaaacgac ggccagtgag ttcgagcttg catgcctgca ggtcgttaca taacttacgg 61 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt 121 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac 181 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg 241 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact 301 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt 361 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc 421 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc 481 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata 541 taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg 601 acctccatag aagacaccgg gaccgatcca gcctccggac tctagaggat ccggtactag 661 aggaactgaa aaaccagaaa gttaactggt aagtttagtc tttttgtctt ttatttcagg 721 tcccggatcc ggtggtggtg caaatcaaag aactgctcct cagtggatgt tgcctttact 781 tctaggcctg tacggaagtg ttacttctgc tctaaaagct gcggaattgt acccgcgggc 841 ccaccatgta cccatacgat gttccagatt acgctcttat ggccatggag gcccgaattc 901 ggtcgaccga gatctctcga ggtacccgtc cggcgtgcaa aatcccgaac gacctgaaac 961 agaaagtcat gaaccacgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc 1021 acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg 1081 gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc aaactgagca 1141 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga 1201 tcactctcgg catggacgag ctgtacaagt aagcggccgc ggggatccag acatgataag 1261 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg 1321 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa 1381 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggttttttc 1441 ggatcctcta gagtcgatct gcaggcatgc tagcttggcg taatcatggt catagctgtt 1501 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa 1561 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact 1621 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc 1681 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg 1741 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc 1801 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag 1861 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 1921 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 1981 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 2041 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag 2101 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 2161 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 2221 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 2281 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt 2341 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 2401 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg 2461 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 2521 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta 2581 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg 2641 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg 2701 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc 2761 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc 2821 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc 2881 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag 2941 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat 3001 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg 3061 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt 3121 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag 3181 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg 3241 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt 3301 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct 3361 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac 3421 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat 3481 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat 3541 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca 3601 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat 3661 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt 3721 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct 3781 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg 3841 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg 3901 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt 3961 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct 4021 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc 4081 acgacgtt //