LOCUS       sequence-436539-        5422 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of Bcl-2-G101V tagged with GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5422)
  AUTHORS   Wang J, Heinz M, Han K, Shah VJ, Hasselbeck S, Schwalm MP, Rathore 
            R, Hummer G, Zhou J, Dikic I, Cheng X
  TITLE     Thalidomide derivatives degrade BCL-2 by reprogramming the binding 
            surface of CRBN
  JOURNAL   Cell Reports Physical Science, Volume 5, Issue 5, 101960
REFERENCE   2  (bases 1 to 5422)
  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: "Cell 
            Reports Physical Science, Volume 5, Issue 5, 101960"
COMMENT     Sequence Label: GFP-Bcl-2-G101V
FEATURES             Location/Qualifiers
     source          1..5422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        105..408
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        409..612
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     563..583
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      642..651
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(648..650,651..653,654..1364)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(693..714)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(954..973)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1301..1322
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2254..2375
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2291..2310)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2345..2364
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2382..2837)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2519..2540)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2731..2750
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2864..2968
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2970..3327
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2970..3161
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2993..3013)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3178..3313
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3362..4156
                     /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             3362..4156
                     /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(3416..3435)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4026..4045
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4344..4363)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4388..4435
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4764..5352
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5253..5272
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa
       61 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      121 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      181 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      241 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      301 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
      361 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      421 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      481 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      541 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      601 ataagcagag ctggtttagt gaaccgtcag atccgctagc cgccaccatg gtgagcaagg
      661 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
      721 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
      781 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct
      841 tgacctacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
      901 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
      961 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     1021 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     1081 actacaacag ccacaaggtc tatatcaccg ccgacaagca gaagaacggc atcaaggtga
     1141 acttcaagac ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     1201 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     1261 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     1321 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagctcaga tctatggcgc
     1381 acgctgggag aacggggtac gataaccggg agatagtgat gaagtacatc cattataagc
     1441 tgtcgcagag gggctacgag tgggatgcgg gagatgtggg cgccgcgccc ccgggggccg
     1501 cccccgcacc gggcatcttc tcctcccagc ccgggcacac gccccatcca gccgcatccc
     1561 gcgacccggt cgccaggacc tcgccgctgc agaccccggc tgcccccggc gccgccgcgg
     1621 ggcctgcgct cagcccggtg ccacctgtgg tccacctgac cctccgccag gccgtggacg
     1681 acttctcccg ccgctaccgc cgcgacttcg ccgagatgtc cagccagctg cacctgacgc
     1741 ccttcaccgc gcggggacgc tttgccacgg tggtggagga gctcttcagg gacggggtga
     1801 actgggggag gattgtggcc ttctttgagt tcggtggggt catgtgtgtg gagagcgtca
     1861 accgggagat gtcgcccctg gtggacaaca tcgccctgtg gatgactgag tacctgaacc
     1921 ggcacctgca cacctggatc caggataacg gaggctggga tgcctttgtg gaactgtacg
     1981 gccccagcat gcggcctctg tttgatttct cctggctgtc tctgaagact ctgctcagtt
     2041 tggccctggt gggagcttgc atcaccctgg gtgcctatct gagccacaag tgagaattct
     2101 gcagtcgacg gtaccgcggg cccgggatcc accggatcta gataactgat cataatcagc
     2161 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     2221 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     2281 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     2341 agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat
     2401 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     2461 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     2521 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     2581 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     2641 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     2701 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     2761 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     2821 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     2881 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     2941 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     3001 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3061 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3121 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3181 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3241 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3301 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     3361 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3421 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3481 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3541 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3601 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3661 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     3721 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     3781 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     3841 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     3901 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     3961 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4021 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4081 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4141 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4201 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4261 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4321 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     4381 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     4441 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     4501 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     4561 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     4621 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4681 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4741 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4801 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4861 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     4921 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4981 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5041 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5101 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5161 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5221 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5281 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5341 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5401 tttgctcaca tgttctttcc tg
//
