LOCUS       sequence-353685-        5432 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express Venus fused to the N-terminus of the Bcl-2 family chimera
            protein, Bcl-2-Cb5: Bcl-2 with its membrane binding region swapped 
            for Cb5 (ER targeting sequence)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5432)
  TITLE     David Andrews Lab plasmids
REFERENCE   2  (bases 1 to 5432)
  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"
COMMENT     Sequence Label: Venus-Bcl2-cb5-pEGFP-C1
FEATURES             Location/Qualifiers
     source          1..5432
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             66..860
                     /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             66..860
                     /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(120..139)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     730..749
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1048..1067)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1092..1139
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1468..2056
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1957..1976
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2231..2534
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2535..2738
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2689..2709
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2777..2786
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2783..2785,2786..2788,2789..3499)
                     /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(2828..2849)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3089..3108)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    4390..4511
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4427..4446)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4481..4500
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4518..4973)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4655..4676)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4867..4886
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5000..5104
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(5106..5432,1..31)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5106..5297
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5129..5149)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(5314..5432,1..17)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
       61 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
      121 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
      181 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
      241 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
      301 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
      361 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
      421 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
      481 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
      541 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
      601 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
      661 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
      721 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
      781 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
      841 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
      901 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
      961 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     1021 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     1081 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     1141 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     1201 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     1261 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     1321 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     1381 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     1441 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     1501 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1561 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     1621 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1681 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1741 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1801 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1861 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1921 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1981 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     2041 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     2101 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     2161 cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     2221 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     2281 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     2341 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     2401 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     2461 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     2521 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     2581 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     2641 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     2701 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc
     2761 gctagcgcta ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
     2821 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     2881 gggcgagggc gatgccacct acggcaagct gaccctgaag ctgatctgca ccaccggcaa
     2941 gctgcccgtg ccctggccca ccctcgtgac caccctgggc tacggcctgc agtgcttcgc
     3001 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     3061 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     3121 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     3181 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     3241 caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga
     3301 ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     3361 cgtgctgctg cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa
     3421 cgagaagcgc gatcacatgg tcctgctgga acgcgtgacc gccgccggga tcactctcgg
     3481 catggacgag ctgtacaagt ccggactcag atctaccatg gcgcacgctg ggagaacagg
     3541 gtacgataac cgggagatag tgatgaagta catccattat aagctgtcgc agaggggcta
     3601 cgagtgggat gcgggagatg tgggcgccgc gcccccgggg gccgcccccg caccgggcat
     3661 cttctcctcc cagcccgggc acacgcccca tccagccgca tcccgggacc cggtcgccag
     3721 gacctcgcca ctgcagaccc cggctgcccc cggcgccgcc gcggggcctg cgctcagccc
     3781 ggtgccacct gtggtccacc tgaccctccg ccaggccggc gacgacttct cccgccgcta
     3841 ccgccgcgac ttcgccgaga tgtccagcca gctgcacctg acgcccttca ccgcgcgggg
     3901 acgctttgcc acggtggtgg aggagctctt cagggacggg gtgaactggg ggaggattgt
     3961 ggccttcttt gagttcggtg gggtcatatg tgtggagagc gtcaaccggg agatgtcgcc
     4021 cctggtggac aacatcgccc tgtggatgac tgagtacctg aaccggcacc tgcacacctg
     4081 gatccaggat aacggaggct gggatgcctt tgtggagctc tacggcccca gcatgcggcc
     4141 tctgtttgat ttctcctggc tgtctcttat cactaccgtt gaatcgaact ctagttggtg
     4201 gactaactgg gttatccctg cgatctctgc tctggttgta gcgctgatgt accgtctgta
     4261 catggctgaa gattaggata actgatcata atcagccata ccacatttgt agaggtttta
     4321 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     4381 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4441 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4501 aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     4561 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     4621 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     4681 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     4741 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     4801 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     4861 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     4921 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     4981 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     5041 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5101 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     5161 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     5221 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5281 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     5341 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     5401 tcggcctctg agctattcca gaagtagtga gg
//
