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 72..375 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 376..579 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 530..550 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 618..627 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 624..1340 /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(669..690) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(930..949) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 1848..1900 /label=MCS /note="multiple cloning site" polyA_signal 2024..2145 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2061..2080) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2115..2134 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2152..2607) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2289..2310) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2501..2520 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2634..2738 /gene="bla" /label=AmpR promoter promoter 2740..3097 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2763..2783) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2948..3083 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3010..3029 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3132..3926 /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(3186..3205) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3796..3815 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4114..4133) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4158..4205 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4534..5122 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5023..5042 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ccgccatgca ttagttatta atagtaatca attacggggt cattagttca tagcccatat 61 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac 121 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc 181 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg 241 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat 301 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc 361 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt 421 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac 481 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc 541 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc 601 cgctagcgct accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg 661 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg 721 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctgatctgc accaccggca 781 agctgcccgt gccctggccc accctcgtga ccaccctggg ctacggcctg cagtgcttcg 841 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct 901 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg 961 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg 1021 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata 1081 tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg 1141 aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc 1201 ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc aaagacccca 1261 acgagaagcg cgatcacatg gtcctgctgg aacgcgtgac cgccgccggg atcactctcg 1321 gcatggacga gctgtacaag tctggactca gatcccgagg aggcatgtct gaagtaagac 1381 ccctctccag agacatcttg atggagaccc tcctgtatga gcagctcctg gaacccccga 1441 ccatggaggt tcttggcatg actgactctg aagaggacct ggaccctatg gaggacttcg 1501 attctttgga atgcatggag ggcagtgacg cagaagcgtt acgagaagcg tgtgaagggg 1561 acgaggagga cgtgagcctc agggccccgc gcctggccca gctctccgag gtggccatgc 1621 acagcctggg tctggctttc atctacgacc agactgagga catcagggat gttcttagaa 1681 gtttcatgga cggtttcacc acacttaagg agaacataat gaggttctgg agatccccga 1741 accccgggtc ctgggtgtcc tgcgaacagg tgctgctggc gctgctgctg ctgctggcgc 1801 tgctgctgcc gctgctcagc gggggcctgc acctgctgct caagtgactc gagctcaagc 1861 ttcgaattct gcagtcgacg gtaccgcggg cccgggatcc accggatcta gataactgat 1921 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct 1981 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc 2041 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 2101 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa 2161 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc 2221 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga 2281 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag 2341 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt 2401 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta 2461 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag 2521 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg 2581 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa 2641 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac 2701 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct 2761 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 2821 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc 2881 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac 2941 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact 3001 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta 3061 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag 3121 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg 3181 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt 3241 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc 3301 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt 3361 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag 3421 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg 3481 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag 3541 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg 3601 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga 3661 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca 3721 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc 3781 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg 3841 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct 3901 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc 3961 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg 4021 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct 4081 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga 4141 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt 4201 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc 4261 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg 4321 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta 4381 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa 4441 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc 4501 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat 4561 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga 4621 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt 4681 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtatcac cgcctacata 4741 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac 4801 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg 4861 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg 4921 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag 4981 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct 5041 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc 5101 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt 5161 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg 5221 tatta //