LOCUS       sequence-429104-        7251 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 7251)
  AUTHORS   Cheng EH, Wei MC, Weiler S, Flavell RA, Mak TW, Lindsten T, 
            Korsmeyer SJ
  TITLE     BCL-2, BCL-X(L) sequester BH3 domain-only molecules preventing BAX- 
            and BAK-mediated mitochondrial apoptosis.
  JOURNAL   Mol Cell 2001 Sep;8(3):705-11.
  PUBMED    11583631
REFERENCE   2  (bases 1 to 7251)
  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: "Mol Cell"; 
            date: "2001-09"; volume: "8(3)"; pages: "705-11"
COMMENT     Sequence Label: 3544 pMIG Bcl-2
FEATURES             Location/Qualifiers
     source          1..7251
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(63..80)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(234..822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(314..333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(993..1784,1785..1853))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1616..1635
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1854..1958)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2026..2044
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2082..2104)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2204..2223
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2417..2439
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2567..2586
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             2786..3302
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    3366..3707
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             3774..4190
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     4120..4142
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     4158..4174
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    4998..5550
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5142..5159)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5369..5388
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(5562..5564,5565..5567,5568..6281)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5607..5628)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5868..5887)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6215..6236
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             6431..6945
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    7107..7123
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(7131..7137,7138..7155,7156..7161))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7176..7197
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
       61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     1861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1981 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     2041 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     2101 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     2161 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     2221 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     2281 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     2341 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     2401 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggcgcaagg aatggtgcat
     2461 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     2521 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     2581 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     2641 agaggcgatt agtccaattt gttaaagaca ggatatcagt ggtccaggct ctagttttga
     2701 ctcaacaata tcaccagctg aagcctatag agtacgagcc atagataaaa taaaagattt
     2761 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
     2821 cttaagtaac gccattttgc aaggcatgga aaatacataa ctgagaatag agaagttcag
     2881 atcaaggtta ggaacagaga gacagcagaa tatgggccaa acaggatatc tgtggtaagc
     2941 agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc ccgccctcag
     3001 cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt
     3061 gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc
     3121 cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc gatagactgc
     3181 gtcgcccggg tacccgtatt cccaataaag cctcttgctg tttgcatccg aatcgtggac
     3241 tcgctgatcc ttgggagggt ctcctcagat tgattgactg cccacctcgg gggtctttca
     3301 tttggaggtt ccaccgagat ttggagaccc ctgcccaggg accaccgacc cccccgccgg
     3361 gaggtaagct ggccagcggt cgtttcgtgt ctgtctctgt ctttgtgcgt gtttgtgccg
     3421 gcatctaatg tttgcgcctg cgtctgtact agttagctaa ctagctctgt atctggcgga
     3481 cccgtggtgg aactgacgag ttctgaacac ccggccgcaa ccctgggaga cgtcccaggg
     3541 actttggggg ccgtttttgt ggcccgacct gaggaaggga gtcgatgtgg aatccgaccc
     3601 cgtcaggata tgtggttctg gtaggagacg agaacctaaa acagttcccg cctccgtctg
     3661 aatttttgct ttcggtttgg aaccgaagcc gcgcgtcttg tctgctgcag cgctgcagca
     3721 tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt gtctgaaaat tagggccaga
     3781 ctgttaccac tcccttaagt ttgaccttag gtcactggaa agatgtcgag cggatcgctc
     3841 acaaccagtc ggtagatgtc aagaagagac gttgggttac cttctgctct gcagaatggc
     3901 caacctttaa cgtcggatgg ccgcgagacg gcacctttaa ccgagacctc atcacccagg
     3961 ttaagatcaa ggtcttttca cctggcccgc atggacaccc agaccaggtc ccctacatcg
     4021 tgacctggga agccttggct tttgaccccc ctccctgggt caagcccttt gtacacccta
     4081 agcctccgcc tcctcttcct ccatccgccc cgtctctccc ccttgaacct cctcgttcga
     4141 ccccgcctcg atcctccctt tatccagccc tcactccttc tctaggcgcc ggaattagat
     4201 ctctcgaggt taacgaattc gcggccgcgg tgagaaccgt taacgtctag aatggcgcac
     4261 gctgggagaa cagggtacga taaccgggag atagtgatga agtacatcca ttataagctg
     4321 tcgcagaggg gctacgagtg ggatgcggga gatgtgggcg ccgcgccccc gggggccgcc
     4381 cccgcaccgg gcatcttctc ctcccagccc gggcacacgc cccatccagc cgcatcccgg
     4441 gacccggtcg ccaggacctc gccgctgcag accccggctg cccccggcgc cgccgcgggg
     4501 cctgcgctca gcccggtgcc acctgtggtc cacctgaccc tccgccaggc cggcgacgac
     4561 ttctcccgcc gctaccgccg cgacttcgcc gagatgtcca gccagctgca cctgacgccc
     4621 ttcaccgcgc ggggacgctt tgccacggtg gtggaggagc tcttcaggga cggggtgaac
     4681 tgggggagga ttgtggcctt ctttgagttc ggtggggtca tgtgtgtgga gagcgtcaac
     4741 cgggagatgt cgcccctggt ggacaacatc gccctgtgga tgactgagta cctgaaccgg
     4801 cacctgcaca cctggatcca ggataacgga ggctgggatg cctttgtgga actgtacggc
     4861 cccagcatgc ggcctctgtt tgatttctcc tggctgtctc tgaagactct gctcagtttg
     4921 gccctggtgg gagcttgcat caccctgggt gcctatctgg gccacaagtg atctacggaa
     4981 tycctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt
     5041 tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc
     5101 tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca
     5161 aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac
     5221 gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg
     5281 ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt
     5341 gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct
     5401 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg
     5461 ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt
     5521 ggttttcctt tgaaaaacac gatgataata tggccacaac catggtgagc aagggcgagg
     5581 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     5641 agttcagcgt gtctggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     5701 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     5761 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     5821 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     5881 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     5941 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     6001 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gcgaacttca
     6061 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     6121 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     6181 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     6241 ccgccgggat cactctcggc atggacgagc tgtacaagta atgaattaat taagaattat
     6301 caccgcttct attcagccag taaggcctgt ctttaatggc ctccggcatg agacactttc
     6361 tagagtcgac ctgcagccaa gcttatcgat aaaataaaag attttattta gtctccagaa
     6421 aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt
     6481 ttgcaaggca tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca
     6541 gagagacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     6601 cagggccaag aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc
     6661 atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa
     6721 ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga
     6781 gcccacaacc cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg
     6841 tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag
     6901 ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcatgggta acagtttctt
     6961 gaagttggag aacaacattc tgagggtagg agtcgaatat taagtaatcc tgactcaatt
     7021 agccactgtt ttgaatccac atactccaat actcctgaaa tagttcatta tggacagcgc
     7081 agaaagagct ggggagaatt gtgaaattgt tatccgctca caattccaca caacatacga
     7141 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     7201 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct g
//
