LOCUS       sequence-210993-        7671 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 7671)
  AUTHORS   Tokuzawa Y, Kaiho E, Maruyama M, Takahashi K, Mitsui K, Maeda M, 
            Niwa H, Yamanaka S
  TITLE     Fbx15 is a novel target of Oct3/4 but is dispensable for embryonic 
            stem cell self-renewal and mouse development.
  JOURNAL   Mol Cell Biol. 2003 Apr . 23(8):2699-708.
  PUBMED    12665572
REFERENCE   2  (bases 1 to 7671)
  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 
            Biol. 2003 Apr . 23(8):2699-708."
COMMENT     Sequence Label: pCAG-Oct3/4-IRES-Neo
FEATURES             Location/Qualifiers
     source          1..7671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          894..1910
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1834..1856
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     intron          3429..3658
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     3641..3660
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3716..4289
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3883..3900)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4110..4129
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4325..5128
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4388..4407)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4998..5017
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5160..5177)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5166..5390
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5437..5456
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5516..5532)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5516..5532)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5529..5551)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5540..5556
                     /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(5564..5570,5571..5588,5589..5594))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5609..5630
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5747..5764)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5918..6506)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5998..6017)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6677..7468,7469..7537))
                     /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     7300..7319
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7538..7642)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc
      661 cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg
      721 ggcggggggg gggggggcgc gcgccagggg gggggggggg gggggggggg gggggggggg
      781 gggggggggg ggggggggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta
      841 tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg
      901 ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc
      961 tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct
     1021 gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt gaaagccttg
     1081 aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg tgcgtgtgtg
     1141 tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag cgctgcgggc
     1201 gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg
     1261 ccccgcggtg cggggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg
     1321 gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca accccccctg cacccccctc
     1381 cccgagttgc tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg
     1441 ggctcgccgt gccgggcggg gggtggcggc aggtgggggt gccgggcggg gcggggccgc
     1501 ctcgggccgg ggagggctcg ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc
     1561 gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac
     1621 ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc accccctcta
     1681 gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg gagggccttc
     1741 gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt ccgcgggggg
     1801 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1861 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1921 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attccgtgag ccgtctttcc
     1981 accaggcccc cggctcgggg tgcccacctt ccccatggct ggcatgcctt cttctttttc
     2041 ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattc
     2101 cgtgagccgt ctttccacca ggcccccggc tcggggtgcc caccttcccc atggctggac
     2161 acctggcttc agacttcgcc ttctcacccc caccaggtgg gggtgatggg tcagcagggc
     2221 tggagccggg ctgggtggat tctcgaacct ggctaagctt ccaagggcct ccaggtgggc
     2281 ctggaatcgg accaggctca gaggtattgg ggatctcccc atgtccgccc gcatacgagt
     2341 tctgcggagg gatggcatac tgtggacctc aggttggact gggcctagtc ccccaagttg
     2401 gcgtggagac tttgcagcct gagggccagg caggagcacg agtggaaagc aactcagagg
     2461 gaacctcctc tgagccctgt gccgaccgcc ccaatgccgt gaagttggag aaggtggaac
     2521 caactcccga ggagtcccag gacatgaaag ccctgcagaa ggagctagaa cagtttgcca
     2581 agctgctgaa gcagaagagg atcaccttgg ggtacaccca ggccgacgtg gggctcaccc
     2641 tgggcgttct ctttggaaag gtgttcagcc agaccaccat ctgtcgcttc gaggccttgc
     2701 agctcagcct taagaacatg tgtaagctgc ggcccctgct ggagaagtgg gtggaggaag
     2761 ccgacaacaa tgagaacctt caggagatat gcaaatcgga gaccctggtg caggcccgga
     2821 agagaaagcg aactagcatt gagaaccgtg tgaggtggag tctggagacc atgtttctga
     2881 agtgcccgaa gccctcccta cagcagatca ctcacatcgc caatcagctt gggctagaga
     2941 aggatgtggt tcgagtatgg ttctgtaacc ggcgccagaa gggcaaaaga tcaagtattg
     3001 agtattccca acgagaagag tatgaggcta cagggacacc tttcccaggg ggggctgtat
     3061 cctttcctct gcccccaggt ccccactttg gcaccccagg ctatggaagc ccccacttca
     3121 ccacactcta ctcagtccct tttcctgagg gcgaggcctt tccctctgtt cccgtcactg
     3181 ctctgggctc tcccatgcat tcaaactgag gcaccagccc tccctgggga tgctgtgagc
     3241 caaggcaagg gaggtagaca agagaacctg gagctttggg gttaaattct tttactgagg
     3301 agggattaaa agcacaacag gggtgggggg tgggatgggg aaagaagctc agtgatgctg
     3361 ttgatccact agtaacggcc gccagtgtgc tggaattaat tcgctgtctg cgagggccag
     3421 ctgttggggt gagtactccc tctcaaaagc gggcatgact tctgcgctaa gattgtcagt
     3481 ttccaaaaac gaggaggatt tgatattcac ctggcccgcg gtgatgcctt tgagggtggc
     3541 cgcgtccatc tggtcagaaa agacaatctt tttgttgtca agcttgaggt gtggcaggct
     3601 tgagatctgg ccatacactt gagtgacaat gacatccact ttgcctttct ctccacaggt
     3661 gtccactccc aggtccaact gcaggtcgag catgcatcta gggcggccaa ttccgcccct
     3721 ctcccccccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     3781 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     3841 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     3901 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
     3961 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     4021 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     4081 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     4141 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat
     4201 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
     4261 tggttttcct ttgaaaaaca cgatgataag cttgccacaa cccgggataa ttcctgcagc
     4321 caatatggga tcggccattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4381 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4441 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4501 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4561 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4621 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4681 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4741 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4801 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4861 gcgcatgccc gacggcgatg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4921 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4981 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5041 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5101 ctatcgcctt cttgacgagt tcttctgagg ggatcaattc tctagagctc gctgatcagc
     5161 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     5221 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     5281 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     5341 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     5401 ggaaagaacc agctggggct cgagtgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5461 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     5521 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5581 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5641 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5701 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5761 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5821 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5881 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5941 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6001 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6061 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6121 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6181 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6241 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6301 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6361 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6421 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6481 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6541 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6601 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6661 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6721 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6781 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6841 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6901 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6961 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7021 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7081 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7141 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7201 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7261 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7321 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7381 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7441 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7501 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7561 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7621 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac g
//
