LOCUS       sequence-436540-        5422 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of Bcl-2-K22R 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-K22R
FEATURES             Location/Qualifiers
     source          1..5422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      605..614
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(611..613,614..616,617..1327)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(656..677)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(917..936)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1264..1285
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2217..2338
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2254..2273)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2308..2327
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2345..2800)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2482..2503)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2694..2713
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2827..2931
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2933..3290
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2933..3124
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2956..2976)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3141..3276
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3325..4119
                     /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             3325..4119
                     /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(3379..3398)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3989..4008
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4307..4326)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4351..4398
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4727..5315
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5216..5235
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agccgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
      661 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
      721 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
      781 ctggcccacc ctcgtgacca ccttgaccta cggcgtgcag tgcttcgccc gctaccccga
      841 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
      901 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
      961 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     1021 cctggggcac aagctggagt acaactacaa cagccacaag gtctatatca ccgccgacaa
     1081 gcagaagaac ggcatcaagg tgaacttcaa gacccgccac aacatcgagg acggcagcgt
     1141 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     1201 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     1261 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     1321 gtacaagctc agatctatgg cgcacgctgg gagaacgggg tacgataacc gggagatagt
     1381 gatgaagtac atccattatc ggctgtcgca gaggggctac gagtgggatg cgggagatgt
     1441 gggcgccgcg cccccggggg ccgcccccgc accgggcatc ttctcctccc agcccgggca
     1501 cacgccccat ccagccgcat cccgcgaccc ggtcgccagg acctcgccgc tgcagacccc
     1561 ggctgccccc ggcgccgccg cggggcctgc gctcagcccg gtgccacctg tggtccacct
     1621 gaccctccgc caggccggcg acgacttctc ccgccgctac cgccgcgact tcgccgagat
     1681 gtccagccag ctgcacctga cgcccttcac cgcgcgggga cgctttgcca cggtggtgga
     1741 ggagctcttc agggacgggg tgaactgggg gaggattgtg gccttctttg agttcggtgg
     1801 ggtcatgtgt gtggagagcg tcaaccggga gatgtcgccc ctggtggaca acatcgccct
     1861 gtggatgact gagtacctga accggcacct gcacacctgg atccaggata acggaggctg
     1921 ggatgccttt gtggaactgt acggccccag catgcggcct ctgtttgatt tctcctggct
     1981 gtctctgaag actctgctca gtttggccct ggtgggagct tgcatcaccc tgggtgccta
     2041 tctgagccac aagtgagaat tctgcagtcg acggtaccgc gggcccggga tccaccggat
     2101 ctagataact gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     2161 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     2221 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     2281 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac
     2341 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     2401 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     2461 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     2521 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     2581 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     2641 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     2701 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     2761 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     2821 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2881 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     2941 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3001 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3061 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3121 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3181 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     3241 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     3301 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     3361 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3421 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3481 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     3541 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3601 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     3661 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3721 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     3781 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     3841 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     3901 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     3961 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4021 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4081 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4141 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4201 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4261 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     4321 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     4381 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     4441 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     4501 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     4561 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4621 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4681 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4741 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4801 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4861 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4921 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4981 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5041 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5101 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5161 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5221 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5281 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5341 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     5401 ctgtggataa ccgtattacc gc
//
