LOCUS       sequence-97397-e        8168 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for constitutive expression of Mbd3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8168)
  AUTHORS   Rais Y, Zviran A, Geula S, Gafni O, Chomsky E, Viukov S, Mansour AA,
            Caspi I, Krupalnik V, Zerbib M, Maza I, Mor N, Baran D, Weinberger 
            L, Jaitin DA, Lara-Astiaso D, Blecher-Gonen R, Shipony Z, Mukamel Z,
            Hagai T, Gilad S, Amann-Zalcenstein D, Tanay A, Amit I, Novershtern 
            N, Hanna JH
  TITLE     Deterministic direct reprogramming of somatic cells to pluripotency.
  JOURNAL   Nature. 2013 Oct 3;502(7469):65-70. doi: 10.1038/nature12587. Epub 
            2013 Sep 18.
  PUBMED    24048479
REFERENCE   2  (bases 1 to 8168)
  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: 
            "10.1038/nature12587"; journalName: "Nature"; date: "2013-10-3- 3"; 
            volume: "502"; issue: "7469"; pages: "65-70"
COMMENT     Sequence Label: pBRPy CAGGS-Mbd3-IRES-PURO
FEATURES             Location/Qualifiers
     source          1..8168
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    576..710
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        724..1107
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1112..1389
                     /label=chicken beta-actin promoter
     intron          1390..2406
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2330..2352
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2414..2433
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_feature    join(3388..3962,3963..3965,3966..3974)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3555..3572)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3782..3801
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3975..4574
                     /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(3975..3994)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4471..4491
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(4655..4672)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4661..4885
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4932..4948)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4932..4948)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4945..4967)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4956..4972
                     /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(4980..4986,4987..5004,5005..5010))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5025..5046
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5105..5304
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5151..5290
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5310..5444
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             5913..6104
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(6098..6120)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6206..6346
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6261..6280
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6361..6378)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6532..7120)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6612..6631)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7291..8082,8083..8151))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7914..7933
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gaccgctttg gaaaaacaaa gactgtattt cctggaaatt aatgtttatt caataaactg
       61 tgtattcagc tatattcaca tagtggtgag gctgaaatga ggcgggaaga ggcggttggg
      121 gcttaattat atcaatttgg gtggccccac agcgcctcca aggcgccagt cctgttttga
      181 caagttgcct ctggaagcct ctcttctttt tctccagagt aagcggaggc caggggcccc
      241 cggcctctgc ttaatactaa aaaaaacagc tgttgtcata gtaatgattg ggtggaaaca
      301 ttccaggcct gggtggagag gctttttgct tcctcttgca aaaccacact gacattccag
      361 gcctgggtgg agaggctttt tgcttcctct tgcaaaacca cactgccctc tggagggcag
      421 ttgcctagca actaattaaa agaggatgtc gcacggccag ctgcggtcag ttagtcactt
      481 cctgcttaac tgacttgaca ttttctattt taagagtcgg gaggaaaatt actgtgttgg
      541 aggccctccg ccatcttctg aagctgaatc gaattaactt gtttattgca gcttataatg
      601 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
      661 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc tgatatcatc
      721 gtcgacattg attattgact agctagttat taatagtaat caattacggg gtcattagtt
      781 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      841 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      901 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      961 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     1021 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     1081 tacgtattag tcatcgctat taccatgatg gtcgaggtga gccccacgtt ctgcttcact
     1141 ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt
     1201 tgtgcagcga tgggggcggg gggggggggg gggcgcgcgc caggcggggc ggggcggggc
     1261 gaggggcggg gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc
     1321 cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg
     1381 cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg
     1441 cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc
     1501 ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg
     1561 ctgcgtgaaa gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg
     1621 ggtgcgtgcg tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc
     1681 tgtgagcgct gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc
     1741 gcggccgggg gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg
     1801 cggggtgtgt gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc
     1861 cccctgcacc cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt
     1921 acggggcgtg gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg
     1981 ggcggggcgg ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggcccccgg
     2041 agcgccggcg gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc
     2101 gagagggcgc agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc
     2161 cgccgcaccc cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg
     2221 ggcggggagg gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg
     2281 ggctgtccgc ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc
     2341 tggcgtgtga ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc
     2401 ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattc
     2461 atggactaca aggacgacga caaggactac aaggacgacg acaagggcag cggaatggag
     2521 cggaagaggt gggagtgccc ggcgctcccg cagggctggg aaagggaaga agtgcccagg
     2581 aggtcggggc tgtcggccgg ccacagggat gtcttttact atagccccag cgggaagaag
     2641 ttccgcagca agccacaact ggcacgttac ctgggcggat ccatggacct cagcaccttc
     2701 gacttccgca ccggaaagat gttgatgaac aagatgaata agagtcgcca gcgtgtgcgc
     2761 tatgattctt ccaaccaggt caagggcaag cctgacctga acaccgcgct gcctgtacgg
     2821 cagactgcat ccatcttcaa gcaaccggtg accaagatca ccaaccaccc cagcaacaag
     2881 gtcaagagcg acccgcagaa ggcagtggac cagccgaggc agcttttctg ggagaagaag
     2941 ctaagtggat tgagtgcctt tgacattgca gaagaactgg tcaggaccat ggacttgccc
     3001 aagggcctgc agggagtggg ccctggctgt acagatgaga cgctgctgtc agccattgcg
     3061 agtgctctac acaccagcac cctgcccatt acaggccagc tctctgcagc cgtggagaag
     3121 aaccctggtg tgtggctgaa cactgcacag ccactgtgca aagccttcat ggtgacagat
     3181 gacgacatca ggaagcagga ggagctggta cagcaggtac ggaagcgcct ggaggaggca
     3241 ctgatggccg acatgctagc tcatgtggag gagcttgccc gagacgggga ggcaccactg
     3301 gacaaggcct gtgcagagga ggaagaggag gaggaagagg aggaggaaga gccggagcca
     3361 gagcgagtgt aggaattccg cccccccccc ctctccctcc ccccccccta acgttactgg
     3421 ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt
     3481 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc
     3541 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc
     3601 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg
     3661 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc
     3721 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa
     3781 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg
     3841 tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa
     3901 aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata
     3961 atatggccac aaccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
     4021 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
     4081 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
     4141 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
     4201 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
     4261 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
     4321 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
     4381 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
     4441 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
     4501 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
     4561 agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac
     4621 cccatgcatc gatgatctag agctcgctga tcagcctcga ctgtgccttc tagttgccag
     4681 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     4741 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     4801 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     4861 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaacctgcag cccaagcttg
     4921 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4981 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5041 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg
     5101 atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     5161 taactccgcc cagttccgcc cattctccgc cccatgcatg gctgactaat tttttttatt
     5221 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     5281 tttggaggcc taggcttttg caaaaagcta acttgtttat tgcagcttat aatggttaca
     5341 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5401 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcccacgg gtgcgcatga
     5461 tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact ggttagcaga
     5521 atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct
     5581 gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt
     5641 cagcgccctg caccattatg ttccggatct gcatcgcagg atgctgctgg ctaccctgtg
     5701 gaacacctac atctgtatta acgaagcgct ggcattgacc ctgagtgatt tttctctggt
     5761 cccgccgcat ccataccgcc agttgtttac cctcacaacg ttccagtaac cgggcatgtt
     5821 catcatcagt aacccgtatc gtgagcatcc tctctcgttt catcggtatc attaccccca
     5881 tgaacagaaa tcccccttac acggaggcat cagtgaccaa acaggaaaaa accgccctta
     5941 acatggcccg ctttatcaga agccagacat taacgcttct ggagaaactc aacgagctgg
     6001 acgcggatga acaggcagac atctgtgaat cgcttcacga ccacgctgat gagctttacc
     6061 gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     6121 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     6181 cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag
     6241 tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     6301 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc
     6361 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6421 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6481 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6541 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6601 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6661 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6721 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6781 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6841 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6901 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6961 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7021 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7081 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7141 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7201 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7261 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7321 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7381 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7441 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7501 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7561 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt
     7621 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7681 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7741 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7801 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7861 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7921 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7981 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8041 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8101 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8161 ttttcaat
//
