LOCUS       sequence-328843-        5318 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To examine effect of swapping the BH3 region in full length BimEL 
            with that of Noxa.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5318)
  AUTHORS   Osterlund EJ, Hirmiz N, Pemberton JM, Nougarede A, Liu Q, Leber B, 
            Fang Q, Andrews DW
  TITLE     Efficacy and specificity of inhibitors of BCL-2 family protein 
            interactions assessed by affinity measurements in live cells.
  JOURNAL   Sci Adv. 2022 Apr 22;8(16):eabm7375. doi: 10.1126/sciadv.abm7375. 
            Epub 2022 Apr 20.
  PUBMED    35442739
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: "Sci Adv."; 
            date: "2022-04-22"; volume: "8(16):eabm7375. doi"; pages: " 
            10.1126/sciadv.abm7375. Epub 2022 Apr 20"
COMMENT     Sequence Label: Venus-BimEL(Noxa)-pEGFP-C1
FEATURES             Location/Qualifiers
     source          1..5318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        154..457
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        458..661
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     612..632
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      700..709
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(706..708,709..711,712..1422)
                     /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
                     GNYETRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     ERVTAAGITLGMDELYK"
     primer_bind     complement(751..772)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    2023..2075
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2199..2320
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2236..2255)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2290..2309
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2327..2782)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2464..2485)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2676..2695
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2809..2913
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2915..3272
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2915..3106
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2938..2958)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3123..3258
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3307..4101
                     /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             3307..4101
                     /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(3361..3380)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3971..3990
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4289..4308)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4333..4380
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4709..5297
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5198..5217
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
       61 gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat caattacggg
      121 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      181 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      241 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      301 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      361 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      421 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
      481 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      541 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
      601 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      661 tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg ccaccatggt gagcaagggc
      721 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
      781 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
      841 aagctgatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
      901 ggctacggcc tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
      961 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     1021 aactacgaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     1081 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     1141 tacaacagcc acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac
     1201 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag
     1261 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag
     1321 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggaacgcgtg
     1381 accgccgccg ggatcactct cggcatggac gagctgtaca agggaggctc catggccaag
     1441 caaccttctg atgtaagttc tgagtgtgac agagaaggtg gacaattgca gcctgctgag
     1501 aggcctcccc agctcaggcc tggggcccct acctccctac agacagaacc gcaaggtaat
     1561 cccgacggcg aaggggaccg ctgcccccac ggcagccctc agggcccgct ggccccaccg
     1621 gccagccctg gcccttttgc taccagatcc ccacttttca tctttgtgag aagatcttct
     1681 ctgctgtccc ggtcctccag tgggtatttc tcttttgaca cagacaggag cccggcaccc
     1741 atgagttgtg acaagtcaac acaaacccca agtcctcctt gccaggcctt caaccactat
     1801 ctcagtgcaa tggcttccat acgacagtct caggaggaac ctgaagatgc agagctggaa
     1861 gtcgagtgtg ctactcaact caggagattt ggagacaaac tgaacttcac ttacacaagg
     1921 agggtgtttg caaatgatta ccgcgaggct gaagaccacc ctcaaatggt tatcttacaa
     1981 ctgttacgct ttatcttccg tctggtatgg agaaggcatt aactcgagct caagcttcga
     2041 attctgcagt cgacggtacc gcgggcccgg gatccaccgg atctagataa ctgatcataa
     2101 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     2161 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     2221 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     2281 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt
     2341 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     2401 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     2461 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     2521 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     2581 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     2641 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     2701 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     2761 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2821 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2881 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     2941 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3001 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3061 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3121 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3181 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3241 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     3301 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3361 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3421 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3481 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3541 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3601 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3661 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3721 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3781 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     3841 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     3901 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     3961 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4021 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4081 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4141 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4201 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4261 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     4321 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     4381 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     4441 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     4501 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     4561 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4621 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4681 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4741 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4801 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4861 tagtgtagcc gtagttaggc caccacttca agaactctgt atcaccgcct acatacctcg
     4921 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4981 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5041 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5101 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5161 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5221 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5281 ggcggagcct atggaaaaac gccagcaacg cggccttt
//
