LOCUS       sequence-328998-        5318 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Venus fused to the N-terminus of the Bcl-2 family protein, 
            BimEL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5318)
  AUTHORS   Liu Q, Osterlund EJ, Chi X, Pogmore J, Leber B, Andrews DW
  TITLE     Bim escapes displacement by BH3-mimetic anti-cancer drugs by 
            double-bolt locking both Bcl-XL and Bcl-2.
  JOURNAL   Elife. 2019 Mar 12;8. pii: 37689. doi: 10.7554/eLife.37689.
  PUBMED    30860026
REFERENCE   2  (bases 1 to 5318)
  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: "Elife."; 
            date: "2019-03-12"; volume: "8. pii: 37689. doi"; pages: " 
            10.7554/eLife.37689"
COMMENT     Sequence Label: Venus-BimEL-pEGFP-C1
FEATURES             Location/Qualifiers
     source          1..5318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        93..396
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        397..600
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     551..571
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      639..648
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(645..647,648..650,651..1361)
                     /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(690..711)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(951..970)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    1962..2014
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2138..2259
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2175..2194)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2229..2248
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2266..2721)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2403..2424)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2615..2634
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2748..2852
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2854..3211
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2854..3045
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2877..2897)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3062..3197
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3246..4040
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3246..4040
                     /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(3300..3319)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3910..3929
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4228..4247)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4272..4319
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4648..5236
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5137..5156
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg
       61 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      121 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      181 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      241 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      301 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      361 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      421 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      481 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      541 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      601 ggtttagtga accgtcagat ccgctagcgc taccggtcgc caccatggtg agcaagggcg
      661 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
      721 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
      781 agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg
      841 gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
      901 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
      961 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     1021 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     1081 acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact
     1141 tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga
     1201 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt
     1261 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gaacgcgtga
     1321 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gggaggctcc atggccaagc
     1381 aaccttctga tgtaagttct gagtgtgaca gagaaggtgg acaattgcag cctgctgaga
     1441 ggcctcccca gctcaggcct ggggccccta cctccctaca gacagaaccg caaggtaatc
     1501 ccgacggcga aggggaccgc tgcccccacg gcagccctca gggcccgctg gccccaccgg
     1561 ccagccctgg cccttttgct accagatccc cacttttcat ctttgtgaga agatcttctc
     1621 tgctgtcccg gtcctccagt gggtatttct cttttgacac agacaggagc ccggcaccca
     1681 tgagttgtga caagtcaaca caaaccccaa gtcctccttg ccaggccttc aaccactatc
     1741 tcagtgcaat ggcttccata cgacagtctc aggaggaacc tgaagatctg cgcccggaga
     1801 tacggattgc acaggagctg cggcggatcg gagacgagtt caacgaaact tacacaagga
     1861 gggtgtttgc aaatgattac cgcgaggctg aagaccaccc tcaaatggtt atcttacaac
     1921 tgttacgctt tatcttccgt ctggtatgga gaaggcatta actcgagctc aagcttcgaa
     1981 ttctgcagtc gacggtaccg cgggcccggg atccaccgga tctagataac tgatcataat
     2041 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     2101 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     2161 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     2221 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta
     2281 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     2341 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     2401 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     2461 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     2521 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     2581 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     2641 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     2701 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2761 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2821 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     2881 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2941 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3001 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3061 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3121 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     3181 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     3241 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3301 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     3361 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     3421 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     3481 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     3541 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     3601 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     3661 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     3721 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     3781 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     3841 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     3901 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     3961 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4021 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     4081 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4141 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4201 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     4261 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     4321 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     4381 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     4441 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     4501 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4561 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4621 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4681 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4741 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     4801 agtgtagccg tagttaggcc accacttcaa gaactctgta tcaccgccta catacctcgc
     4861 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4921 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4981 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5041 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5101 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5161 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5221 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5281 gccttttgct cacatgttct ttcctgcgtt atcccctg
//
