LOCUS       sequence-159452-        5084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5084)
  AUTHORS   Wang NS, Unkila MT, Reineks EZ, Distelhorst CW
  TITLE     Transient expression of wild-type or mitochondrially targeted Bcl-2 
            induces apoptosis, whereas transient expression of endoplasmic 
            reticulum-targeted Bcl-2 is protective against Bax-induced cell 
            death.
  JOURNAL   J Biol Chem. 2001 Nov 23. 276(47):44117-28.
  PUBMED    11546793
REFERENCE   2  (bases 1 to 5084)
  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: "J Biol 
            Chem. 2001 Nov 23. 276(47):44117-28."
COMMENT     Sequence Label: Flag-Bcl2
FEATURES             Location/Qualifiers
     source          1..5084
                     /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"
     promoter        621..639
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      674..683
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             683..706
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1462..1478)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1463..1479)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1534..1553)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1535..1553)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1827..1948
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1864..1883)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1918..1937
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1955..2410)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2092..2113)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2304..2323
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2437..2541
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2543..2900
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2543..2734
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2566..2586)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2751..2886
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2935..3729
                     /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             2935..3729
                     /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(2989..3008)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3599..3618
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3917..3936)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3961..4008
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4337..4925
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4826..4845
                     /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 agcgattacg ccaagctcga aattaaccct cactaaaggg aacaaaagct ggagctccac
      661 cgcggtggcg gccgccacca tggattacaa ggatgacgac gataagagcc cgggcggatc
      721 tatggcgcac gctgggagaa cggggtacga taaccgggag atagtgatga agtacatcca
      781 ttataagctg tcgcagaggg gctacgagtg ggatgcggga gatgtgggcg ccgcgccccc
      841 gggggccgcc cccgcaccgg gcatcttctc ctcccagccc gggcacacgc cccatccagc
      901 cgcatcccgc gacccggtcg ccaggacctc gccgctgcag accccggctg cccccggcgc
      961 cgccgcgggg cctgcgctca gcccggtgcc acctgtggtc cacctgaccc tccgccaggc
     1021 cggcgacgac ttctcccgcc gctaccgccg cgacttcgcc gagatgtcca gccagctgca
     1081 cctgacgccc ttcaccgcgc ggggacgctt tgccacggtg gtggaggagc tcttcaggga
     1141 cggggtgaac tgggggagga ttgtggcctt ctttgagttc ggtggggtca tgtgtgtgga
     1201 gagcgtcaac cgggagatgt cgcccctggt ggacaacatc gccctgtgga tgactgagta
     1261 cctgaaccgg cacctgcaca cctggatcca ggataacgga ggctgggatg cctttgtgga
     1321 actgtacggc cccagcatgc ggcctctgtt tgatttctcc tggctgtctc tgaagactct
     1381 gctcagtttg gccctggtgg gagcttgcat caccctgggt gcctatctga gccacaagtg
     1441 agaattcgat atcaagctta tcgataccgt cgacctcgag ggggggcccg gtaccttaat
     1501 taattaaggt accaggtaag tgtacccaat tcgccctata gtgagtcgta ttacaattca
     1561 ctcgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggagatc caatttttaa
     1621 gtgtataatg tgttaaacta ctgattctaa ttgtttgtgt attttagatt cacagtccca
     1681 aggctcattt caggcccctc agtcctcaca gtctgttcat gatcataatc agccatacca
     1741 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     1801 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     1861 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     1921 gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta
     1981 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     2041 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     2101 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     2161 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     2221 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     2281 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     2341 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     2401 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     2461 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     2521 taatattgaa aaaggaagaa tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     2581 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2641 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2701 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     2761 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     2821 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     2881 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     2941 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3001 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3061 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     3121 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3181 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3241 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3301 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3361 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagaacat
     3421 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     3481 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     3541 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     3601 ttggctaccc gtgatattgc tgaagaactt ggcggcgaat gggctgaccg cttcctcgtg
     3661 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     3721 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     3781 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     3841 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     3901 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     3961 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     4021 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     4081 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     4141 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     4201 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4261 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4321 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4381 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4441 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4501 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4561 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4621 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4681 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4741 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4801 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4861 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4921 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4981 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt
     5041 agttattaat agtaatcaat tacggggtca ttagttcata gccc
//
