LOCUS       sequence-328840-        5318 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To examine effect of swapping the BH3 region in full length BimEL 
            with that of Bad.
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(Bad)-pEGFP-C1
FEATURES             Location/Qualifiers
     source          1..5318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        47..350
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        351..554
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     505..525
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      593..602
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(599..601,602..604,605..1315)
                     /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(644..665)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(905..924)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    1916..1968
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2092..2213
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2129..2148)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2183..2202
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2220..2675)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2357..2378)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2569..2588
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2702..2806
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2808..3165
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2808..2999
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2831..2851)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3016..3151
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3200..3994
                     /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             3200..3994
                     /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(3254..3273)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3864..3883
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4182..4201)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4226..4273
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4602..5190
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5091..5110
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
       61 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      121 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      181 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      241 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      301 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      361 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      421 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      481 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      541 atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg tcgccaccat
      601 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
      661 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
      721 caagctgacc ctgaagctga tctgcaccac cggcaagctg cccgtgccct ggcccaccct
      781 cgtgaccacc ctgggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca
      841 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
      901 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
      961 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1021 gctggagtac aactacaaca gccacaacgt ctatatcacc gccgacaagc agaagaacgg
     1081 catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga
     1141 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1201 cctgagctac cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1261 gctggaacgc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggagg
     1321 ctccatggcc aagcaacctt ctgatgtaag ttctgagtgt gacagagaag gtggacaatt
     1381 gcagcctgct gagaggcctc cccagctcag gcctggggcc cctacctccc tacagacaga
     1441 accgcaaggt aatcccgacg gcgaagggga ccgctgcccc cacggcagcc ctcagggccc
     1501 gctggcccca ccggccagcc ctggcccttt tgctaccaga tccccacttt tcatctttgt
     1561 gagaagatct tctctgctgt cccggtcctc cagtgggtat ttctcttttg acacagacag
     1621 gagcccggca cccatgagtt gtgacaagtc aacacaaacc ccaagtcctc cttgccaggc
     1681 cttcaaccac tatctcagtg caatggcttc catacgacag tctcaggagg aacctgaaga
     1741 tctctgggca gcacagcgct atggccgcga gctccggagg atgagtgacg agtttgtgga
     1801 cacttacaca aggagggtgt ttgcaaatga ttaccgcgag gctgaagacc accctcaaat
     1861 ggttatctta caactgttac gctttatctt ccgtctggta tggagaaggc attaactcga
     1921 gctcaagctt cgaattctgc agtcgacggt accgcgggcc cgggatccac cggatctaga
     1981 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     2041 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     2101 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2161 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta
     2221 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     2281 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     2341 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     2401 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     2461 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     2521 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     2581 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     2641 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     2701 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2761 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     2821 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2881 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     2941 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3001 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3061 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3121 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     3181 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3241 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3301 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3361 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     3421 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     3481 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     3541 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     3601 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     3661 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     3721 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     3781 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     3841 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     3901 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     3961 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     4021 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     4081 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4141 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     4201 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     4261 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     4321 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     4381 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     4441 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4501 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4561 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4621 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4681 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4741 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtatcaccg
     4801 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4861 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4921 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4981 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5041 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5101 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5161 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5221 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5281 gataaccgta ttaccgccat gcattagtta ttaatagt
//
