LOCUS       sequence-134938-        7236 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 7236)
  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 7236)
  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: 3541 pMIG Bcl-XL
FEATURES             Location/Qualifiers
     source          1..7236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1974..1992
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2030..2052)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2152..2171
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2365..2387
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2515..2534
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             2734..3250
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    3314..3655
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             3722..4138
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     4068..4090
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     4106..4122
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    4931..5483
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5075..5092)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5302..5321
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(5495..5497,5498..5500,5501..6214)
                     /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(5540..5561)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5801..5820)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6148..6169
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             6364..6878
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    7040..7056
                     /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(7064..7070,7071..7088,7089..7094))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7109..7130
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     1981 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     2041 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     2101 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     2161 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     2221 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     2281 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     2341 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gcgcaaggaa
     2401 tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc
     2461 cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
     2521 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
     2581 tccggcgtag aggcgattag tccaatttgt taaagacagg atatcagtgg tccaggctct
     2641 agttttgact caacaatatc accagctgaa gcctatagag tacgagccat agataaaata
     2701 aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg
     2761 caagctagct taagtaacgc cattttgcaa ggcatggaaa atacataact gagaatagag
     2821 aagttcagat caaggttagg aacagagaga cagcagaata tgggccaaac aggatatctg
     2881 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtccc
     2941 gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat
     3001 gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt
     3061 ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga
     3121 tagactgcgt cgcccgggta cccgtattcc caataaagcc tcttgctgtt tgcatccgaa
     3181 tcgtggactc gctgatcctt gggagggtct cctcagattg attgactgcc cacctcgggg
     3241 gtctttcatt tggaggttcc accgagattt ggagacccct gcccagggac caccgacccc
     3301 cccgccggga ggtaagctgg ccagcggtcg tttcgtgtct gtctctgtct ttgtgcgtgt
     3361 ttgtgccggc atctaatgtt tgcgcctgcg tctgtactag ttagctaact agctctgtat
     3421 ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg
     3481 tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt cgatgtggaa
     3541 tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac agttcccgcc
     3601 tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg
     3661 ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta
     3721 gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag atgtcgagcg
     3781 gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct tctgctctgc
     3841 agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc gagacctcat
     3901 cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag accaggtccc
     3961 ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca agccctttgt
     4021 acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc ttgaacctcc
     4081 tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc taggcgccgg
     4141 aattagatct ctcgaggtta acgaattgtc accatgtctc agagcaaccg ggagctggtg
     4201 gttgactttc tctcctacaa gctttcccag aaaggataca gctggagtca gtttagtgat
     4261 gtggaagaga acaggactga ggccccagaa gggactgaat cggagatgga gacccccagt
     4321 gccatcaatg gcaacccatc ctggcacctg gcagacagcc ccgcggtgaa tggagccact
     4381 ggccacagca gcagtttgga tgcccgggag gtgatcccca tggcagcagt aaagcaagcg
     4441 ctgagggagg caggcgacga gtttgaactg cggtaccggc gggcattcag tgacctgaca
     4501 tcccagctcc acatcacccc agggacagca tatcagagct ttgaacaggt agtgaatgaa
     4561 ctcttccggg atggggtaaa ctggggtcgc attgtggcct ttttctcctt cggcggggca
     4621 ctgtgcgtgg aaagcgtaga caaggagatg caggtattgg tgagtcggat cgcagcttgg
     4681 atggccactt acctgaatga ccacctagag ccttggatcc aggagaacgg cggctgggat
     4741 acttttgtgg aactctatgg gaacaatgca gcagccgaga gccgaaaggg ccaggaacgc
     4801 ttcaaccgct ggttcctgac gggcatgact gtggccggcg tggttctgct gggctcactc
     4861 ttcagtcgga aatgaccaga cactgaccat ccactctacg gtcacaattc cgccccctcc
     4921 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     4981 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     5041 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     5101 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     5161 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     5221 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     5281 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     5341 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca
     5401 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     5461 ctttgaaaaa cacgatgata atatggccac aaccatggtg agcaagggcg aggagctgtt
     5521 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     5581 cgtgtctggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     5641 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     5701 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     5761 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     5821 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     5881 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     5941 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggcgaact tcaagatccg
     6001 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     6061 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     6121 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     6181 gatcactctc ggcatggacg agctgtacaa gtaatgaatt aattaagaat tatcaccgct
     6241 tctattcagc cagtaaggcc tgtctttaat ggcctccggc atgagacact ttctagagtc
     6301 gacctgcagc caagcttatc gataaaataa aagattttat ttagtctcca gaaaaagggg
     6361 ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag
     6421 gcatggaaaa tacataactg agaatagaga agttcagatc aaggttagga acagagagac
     6481 agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     6541 aagaacagat ggtccccaga tgcggtcccg ccctcagcag tttctagaga accatcagat
     6601 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     6661 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
     6721 acccctcact cggcgcgcca gtcctccgat agactgcgtc gcccgggtac ccgtgtatcc
     6781 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
     6841 tctgagtgat tgactacccg tcagcggggg tctttcatgg gtaacagttt cttgaagttg
     6901 gagaacaaca ttctgagggt aggagtcgaa tattaagtaa tcctgactca attagccact
     6961 gttttgaatc cacatactcc aatactcctg aaatagttca ttatggacag cgcagaaaga
     7021 gctggggaga attgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     7081 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     7141 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     7201 aacgcgcggg gagaggcggt ttgcgtattg ggcgct
//
