LOCUS Exported 5225 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Express Venus fused to the N-terminus of the Bcl-2 family protein, Bik with mutation in BH3 domain to disrupt binding anti-apoptotic proteins. ACCESSION . VERSION . KEYWORDS Venus-Bik-4E-pEGFP-C1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5225) TITLE Andrew lab Venus and Cerulean fusions REFERENCE 2 (bases 1 to 5225) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5225 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 607..616 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 613..1329 /codon_start=1 /product="yellow fluorescent protein (YFP) with fast and efficient maturation (Nagai et al., 2002)" /label=Venus /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL ERVTAAGITLGMDELYK" primer_bind complement(658..679) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(919..938) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 1837..1889 /label=MCS /note="multiple cloning site" polyA_signal 2013..2134 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2050..2069) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2104..2123 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2141..2596) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2278..2299) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2490..2509 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2623..2727 /gene="bla" /label=AmpR promoter promoter 2729..3086 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2752..2772) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2937..3072 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2999..3018 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3121..3915 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3175..3194) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3785..3804 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4103..4122) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4147..4194 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4523..5111 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5012..5031 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg 661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc 721 gatgccacct acggcaagct gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg 781 ccctggccca ccctcgtgac caccctgggc tacggcctgc agtgcttcgc ccgctacccc 841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag 901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag 961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac 1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac 1081 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc 1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg 1201 cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc 1261 gatcacatgg tcctgctgga acgcgtgacc gccgccggga tcactctcgg catggacgag 1321 ctgtacaagt ctggactcag atcccgagga ggcatgtctg aagtaagacc cctctccaga 1381 gacatcttga tggagaccct cctgtatgag cagctcctgg aacccccgac catggaggtt 1441 cttggcatga ctgactctga agaggacctg gaccctatgg aggacttcga ttctttggaa 1501 tgcatggagg gcagtgacgc agaagcgtta cgagaagcgt gtgaagggga cgaggaggac 1561 gtgagcctca gggccccgcg cctggcccag ctctccgagg tggccatgca cagcctgggt 1621 ctggctttca tctacgacca gactgaggac atcagggatg ttcttagaag tttcatggac 1681 ggtttcacca cacttaagga gaacataatg aggttctgga gatccccgaa ccccgggtcc 1741 tgggtgtcct gcgaacaggt gctgctggcg ctgctgctgc tgctggcgct gctgctgccg 1801 ctgctcagcg ggggcctgca cctgctgctc aagtgactcg agctcaagct tcgaattctg 1861 cagtcgacgg taccgcgggc ccgggatcca ccggatctag ataactgatc ataatcagcc 1921 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc 1981 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt 2041 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 2101 gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt 2161 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa 2221 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca 2281 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc 2341 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg 2401 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg 2461 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg 2521 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg 2581 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt 2641 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg 2701 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt 2761 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 2821 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta 2881 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc 2941 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta 3001 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct 3061 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc 3121 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc 3181 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca 3241 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg 3301 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg 3361 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag 3421 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg 3481 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc 3541 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa 3601 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac 3661 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat 3721 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac 3781 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc 3841 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt 3901 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc 3961 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg 4021 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg 4081 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg 4141 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac 4201 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc 4261 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag 4321 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac 4381 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg 4441 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 4501 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 4561 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 4621 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt 4681 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 4741 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 4801 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 4861 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 4921 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 4981 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 5041 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 5101 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 5161 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca 5221 tgcat //