LOCUS       sequence-421245-        8098 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 8098)
  AUTHORS   Maruyama M, Ichisaka T, Nakagawa M, Yamanaka S
  TITLE     Differential roles for Sox15 and Sox2 in transcriptional control in 
            mouse embryonic stem cells.
  JOURNAL   J Biol Chem. 2005 Jul 1. 280(26):24371-9.
  PUBMED    15863505
REFERENCE   2  (bases 1 to 8098)
  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. 2005 Jul 1. 280(26):24371-9."
COMMENT     Sequence Label: pCAG-Myc-Oct3/4-IP
FEATURES             Location/Qualifiers
     source          1..8098
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          227..1243
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1167..1189
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1251..1270
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1318..1347
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1357..1386
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1396..1425
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1435..1464
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1474..1503
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1537..1566
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    1576..1600
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2829..2853)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3036..3588
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3180..3197)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3407..3426
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3600..4199
                     /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(3600..3619)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4096..4116
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(4280..4297)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4286..4510
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4557..4573)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4557..4573)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4570..4592)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4581..4597
                     /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(4605..4611,4612..4629,4630..4635))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4650..4671
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4730..4925
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4776..4911
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4931..5065
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5133..5150)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5304..5892)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5384..5403)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6063..6854,6855..6923))
                     /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     6686..6705
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    7516..7650
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        7664..8043
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(8045..8098,1..226)
                     /label=chicken beta-actin promoter
ORIGIN
        1 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg
       61 gggggggggg cgsgcgccrg gsggggcggg gcggggcgag gggcggggcg gggcgaggcg
      121 gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag
      181 gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg
      241 ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact
      301 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta
      361 gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct
      421 ccgggagggc cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt
      481 ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc
      541 ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg
      601 gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg
      661 agcagggggt gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc ctccccgagt
      721 tgctgagcac ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc
      781 cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc
      841 cggggagggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc
      901 ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg gacttccttt
      961 gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg
     1021 cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc
     1081 gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg gggacggctg
     1141 ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag
     1201 agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct
     1261 ggttattgtg ctgtctcatc attttggcaa agaattcctc gacgatttaa agctatggag
     1321 caaaagctca tttctgaaga ggacttgaat gaaatggagc aaaagctcat ttctgaagag
     1381 gacttgaatg aaatggagca aaagctcatt tctgaagagg acttgaatga aatggagcaa
     1441 aagctcattt ctgaagagga cttgaatgaa atggagcaaa agctcatttc tgaagaggac
     1501 ttgaatgaaa tggagagctt gggcgacctc accatggagc aaaagctcat ttctgaagag
     1561 gacttgaatt atcaaacaag tttgtacaaa aaagcaggct ccaccttccc catggctgga
     1621 cacctggctt cagacttcgc cttctcaccc ccaccaggtg ggggtgatgg gtcagcaggg
     1681 ctggagccgg gctgggtgga ttctcgaacc tggctaagct tccaagggcc tccaggtggg
     1741 cctggaatcg gaccaggctc agaggtattg gggatctccc catgtccgcc cgcatacgag
     1801 ttctgcggag ggatggcata ctgtggacct caggttggac tgggcctagt cccccaagtt
     1861 ggcgtggaga ctttgcagcc tgagggccag gcaggagcac gagtggaaag caactcagag
     1921 ggaacctcct ctgagccctg tgccgaccgc cccaatgccg tgaagttgga gaaggtggaa
     1981 ccaactcccg aggagtccca ggacatgaaa gccctgcaga aggagctaga acagtttgcc
     2041 aagctgctga agcagaagag gatcaccttg gggtacaccc aggccgacgt ggggctcacc
     2101 ctgggcgttc tctttggaaa ggtgttcagc cagaccacca tctgtcgctt cgaggccttg
     2161 cagctcagcc ttaagaacat gtgtaagctg cggcccctgc tggagaagtg ggtggaggaa
     2221 gccgacaaca atgagaacct tcaggagata tgcaaatcgg agaccctggt gcaggcccgg
     2281 aagagaaagc gaactagcat tgagaaccgt gtgaggtgga gtctggagac catgtttctg
     2341 aagtgcccga agccctccct acagcagatc actcacatcg ccaatcagct tgggctagag
     2401 aaggatgtgg ttcgagtatg gttctgtaac cggcgccaga agggcaaaag atcaagtatt
     2461 gagtattccc aacgagaaga gtatgaggct acagggacac ctttcccagg gggggctgta
     2521 tcctttcctc tgcccccagg tccccacttt ggcaccccag gctatggaag cccccacttc
     2581 accacactct actcagtccc ttttcctgag ggcgaggcct ttccctctgt tcccgtcact
     2641 gctctgggct ctcccatgca ttcaaactga ggcaccagcc ctccctgggg atgctgtgag
     2701 ccaaggcaag ggaggtagac aagagaacct ggagctttgg ggttaaattc ttttactgag
     2761 gagggattaa aagcacaaca ggggtggggg gtgggatggg gaaagaagct cagtgatgct
     2821 gttgatcaac ccagctttct tgtacaaagt ggttcgataa ttcaaggcct ctcgatcgag
     2881 gaattcactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt ggccaactgg
     2941 ctcacaaata ccactgagat cgacggtatc gagcggccgc tcgataagct tgatatcgaa
     3001 ttccgccccc ccccncyctc mctccccccc ccctaacgtt actggccgaa gccgcttgga
     3061 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa
     3121 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
     3181 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc
     3241 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
     3301 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca
     3361 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag
     3421 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct
     3481 ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc
     3541 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg gccacaacca
     3601 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac
     3661 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc
     3721 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     3781 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     3841 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     3901 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     3961 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     4021 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     4081 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     4141 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac
     4201 gcccgcccca cgacccgcag cgcccgaccg aaaggagcgc acgaccccat gcatcgatga
     4261 tctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     4321 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     4381 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     4441 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     4501 ggctctatgg cttctgaggc ggaaagaacc tgcagcccaa gcttggcgta atcatggtca
     4561 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4621 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4681 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt
     4741 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4801 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4861 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4921 gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4981 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5041 caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga
     5101 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5161 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5221 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5281 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5341 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5401 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5461 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5521 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5581 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5641 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5701 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5761 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5821 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5881 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5941 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6001 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6061 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6121 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6181 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6241 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6301 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6361 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6421 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6481 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6541 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6601 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6661 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6721 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6781 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6841 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6901 acacggaaat gttgaatact catactcttc ctttttcaat gaccgctttg gaaaaacaaa
     6961 gactgtattt cctggaaatt aatgtttatt caataaactg tgtattcagc tatattcaca
     7021 tagtggtgag gctgaaatga ggcgggaaga ggcggttggg gcttaattat atcaatttgg
     7081 gtggccccac agcgcctcca aggcgccagt cctgttttga caagttgcct ctggaagcct
     7141 ctcttctttt tctccagagt aagcggaggc caggggcccc cggcctctgc ttaatactaa
     7201 aaaaaacagc tgttgtcata gtaatgattg ggtggaaaca ttccaggcct gggtggagag
     7261 gctttttgct tcctcttgca aaaccacact gacattccag gcctgggtgg agaggctttt
     7321 tgcttcctct tgcaaaacca cactgccctc tggagggcag ttgcctagca actaattaaa
     7381 agaggatgtc gcacggccag ctgcggtcag ttagtcactt cctgcttaac tgacttgaca
     7441 ttttctattt taagagtcgg gaggaaaatt actgtgttgg aggccctccg ccatcttctg
     7501 aagctgaatc gaattaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     7561 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     7621 ctcatcaatg tatcttatca tgtctggatc tgatatcatc gtcgacattg attattgact
     7681 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     7741 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     7801 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     7861 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     7921 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     7981 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     8041 atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc ctccccac
//
