LOCUS       sequence-223323-        8166 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral expression of Bcl2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8166)
  AUTHORS   Cheng Z, Gong Y, Ma Y, Lu K, Lu X, Pierce LA, Thompson RC, Muller S,
            Knapp S, Wang J
  TITLE     Inhibition of BET bromodomain targets genetically diverse 
            glioblastoma.
  JOURNAL   Clin Cancer Res. 2013 Apr 1;19(7):1748-59. doi: 
            10.1158/1078-0432.CCR-12-3066. Epub 2013 Feb 12.
  PUBMED    23403638
REFERENCE   2  (bases 1 to 8166)
  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"; doi: 
            "10.1158/1078-0432.CCR-12-3066"; journalName: "Clin Cancer Res"; 
            date: "2013-04-1- 1"; volume: "19"; issue: "7"; pages: "1748-59"
COMMENT     Sequence Label: pCDH-puro-Bcl2
FEATURES             Location/Qualifiers
     source          1..8166
                     /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"
     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     2379..2396
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2380..2396
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2411..2637
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2638..2818
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2865..2990
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3483..3716
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3901..3945
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    4212..4328
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4418..4621
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4572..4592
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4618..4642
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        5525..5736
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             5749..6017
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             6044..6643
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6044..6063)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6540..6560
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6650..7238
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6703..6723)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7121..7132)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7312..7545
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7617..7738
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7654..7673)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7708..7727
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7778..7913
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7946..7962)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7946..7962)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7959..7981)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7970..7986
                     /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(7994..8000,8001..8018,8019..8024))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8039..8060
                     /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 gccagtgcca
     2401 agctgacgcg tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt
     2461 tagcaacatg ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg
     2521 tggtacgatc gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc
     2581 actgaattgc cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg
     2641 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     2701 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     2761 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     2821 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
     2881 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
     2941 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
     3001 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     3061 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     3121 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     3181 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     3241 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     3301 acaaaagtaa gaccaccgca cagcaagcgg ccactgatct tcagacctgg aggaggagat
     3361 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     3421 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     3481 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     3541 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     3601 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     3661 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     3721 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     3781 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     3841 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     3901 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     3961 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     4021 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     4081 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     4141 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg
     4201 tatcggttaa cttttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata
     4261 gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaattc
     4321 aaaattttat cgatactagt attatgccca gtacatgacc ttatgggact ttcctacttg
     4381 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
     4441 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     4501 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     4561 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtttata taagcagagc
     4621 tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag
     4681 aagattctag agctagcgaa ttcgcggccg cggtgagaac cgttaacgtc tagaatggcg
     4741 cacgctggga gaacagggta cgataaccgg gagatagtga tgaagtacat ccattataag
     4801 ctgtcgcaga ggggctacga gtgggatgcg ggagatgtgg gcgccgcgcc cccgggggcc
     4861 gcccccgcac cgggcatctt ctcctcccag cccgggcaca cgccccatcc agccgcatcc
     4921 cgggacccgg tcgccaggac ctcgccgctg cagaccccgg ctgcccccgg cgccgccgcg
     4981 gggcctgcgc tcagcccggt gccacctgtg gtccacctga ccctccgcca ggccggcgac
     5041 gacttctccc gccgctaccg ccgcgacttc gccgagatgt ccagccagct gcacctgacg
     5101 cccttcaccg cgcggggacg ctttgccacg gtggtggagg agctcttcag ggacggggtg
     5161 aactggggga ggattgtggc cttctttgag ttcggtgggg tcatgtgtgt ggagagcgtc
     5221 aaccgggaga tgtcgcccct ggtggacaac atcgccctgt ggatgactga gtacctgaac
     5281 cggcacctgc acacctggat ccaggataac ggaggctggg atgcctttgt ggaactgtac
     5341 ggccccagca tgcggcctct gtttgatttc tcctggctgt ctctgaagac tctgctcagt
     5401 ttggccctgg tgggagcttg catcaccctg ggtgcctatc tgggccacaa gtgatctacg
     5461 gaattcgaat ttaaatcgga tccgcggccg cgaaggatct gcgatcgctc cggtgcccgt
     5521 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
     5581 tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
     5641 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
     5701 gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc
     5761 tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt
     5821 ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag
     5881 ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg
     5941 ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt
     6001 ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagatgaccg agtacaagcc
     6061 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc
     6121 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt
     6181 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     6241 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     6301 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     6361 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     6421 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     6481 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     6541 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     6601 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgagtcgaca atcaacctct
     6661 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     6721 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     6781 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     6841 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     6901 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     6961 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     7021 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     7081 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     7141 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     7201 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggtaccttta agaccaatga
     7261 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
     7321 taattcactc ccaacgaaga taagatctgc tttttgcttg tactgggtct ctctggttag
     7381 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     7441 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     7501 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc
     7561 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact
     7621 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     7681 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     7741 atgtctggct ctagctatcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     7801 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     7861 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagactt
     7921 ttgcagagac caaattcgta atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc
     7981 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     8041 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     8101 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     8161 ggcgct
//
