LOCUS       sequence-199066-        7063 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 7063)
  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 7063)
  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-Sox2-IRES-Neo
FEATURES             Location/Qualifiers
     source          1..7063
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     336..358
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     420..439
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    469..493
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1454..1478)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     intron          1557..1786
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     1769..1788
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    1844..2417
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2011..2028)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2238..2257
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2453..3256
                     /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(2516..2535)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3126..3145
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3288..3305)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3294..3518
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3565..3584
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3644..3660)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3644..3660)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3657..3679)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3668..3684
                     /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(3692..3698,3699..3716,3717..3722))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3737..3758
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3875..3892)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4046..4634)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4126..4145)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4805..5596,5597..5665))
                     /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     5428..5447
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5666..5770)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5802..6181
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6183..6458
                     /label=chicken beta-actin promoter
     intron          join(6459..7063,1..412)
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
ORIGIN
        1 gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg ccggcggctg
       61 tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg
      121 acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc
      181 tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct
      241 tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg
      301 ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg
      361 cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg
      421 caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattatcac aagtttgtac
      481 aaaaaagcag gcttgtataa catgatggag acggagctga agccgccggg cccgcagcaa
      541 gcttcggggg gcggcggcgg aggaggcaac gccacggcgg cggcgaccgg cggcaaccag
      601 aagaacagcc cggaccgcgt caagaggccc atgaacgcct tcatggtatg gtcccggggg
      661 cagcggcgta agatggccca ggagaacccc aagatgcaca actcggagat cagcaagcgc
      721 ctgggcgcgg agtggaaact tttgtccgag accgagaagc ggccgttcat cgacgaggcc
      781 aagcggctgc gcgctctgca catgaaggag cacccggatt ataaataccg gccgcggcgg
      841 aaaaccaaga cgctcatgaa gaaggataag tacacgcttc ccggaggctt gctggccccc
      901 ggcgggaaca gcatggcgag cggggttggg gtgggcgccg gcctgggtgc gggcgtgaac
      961 cagcgcatgg acagctacgc gcacatgaac ggctggagca acggcagcta cagcatgatg
     1021 caggagcagc tgggctaccc gcagcacccg ggcctcaacg ctcacggcgc ggcacagatg
     1081 caaccgatgc accgctacga cgtcagcgcc ctgcagtaca actccatgac cagctcgcag
     1141 acctacatga acggctcgcc cacctacagc atgtcctact cgcagcaggg cacccccggt
     1201 atggcgctgg gctccatggg ctctgtggtc aagtccgagg ccagctccag cccccccgtg
     1261 gttacctctt cctcccactc cagggcgccc tgccaggccg gggacctccg ggacatgatc
     1321 agcatgtacc tccccggcgc cgaggtgccg gagcccgctg cgcccagtag actgcacatg
     1381 gcccagcact accagagcgg cccggtgccc ggcacggcca ttaacggcac actgcccctg
     1441 tcgcacatgt gaaacccagc tttcttgtac aaagtggtga taattcagtg gatccactag
     1501 taacggccgc cagtgtgctg gaattaattc gctgtctgcg agggccagct gttggggtga
     1561 gtactccctc tcaaaagcgg gcatgacttc tgcgctaaga ttgtcagttt ccaaaaacga
     1621 ggaggatttg atattcacct ggcccgcggt gatgcctttg agggtggccg cgtccatctg
     1681 gtcagaaaag acaatctttt tgttgtcaag cttgaggtgt ggcaggcttg agatctggcc
     1741 atacacttga gtgacaatga catccacttt gcctttctct ccacaggtgt ccactcccag
     1801 gtccaactgc aggtcgagca tgcatctagg gcggccaatt ccgcccctct cccccccccc
     1861 cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt
     1921 tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct
     1981 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga
     2041 atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga
     2101 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac
     2161 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag
     2221 ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc
     2281 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg
     2341 tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt
     2401 gaaaaacacg atgataagct tgccacaacc cgggataatt cctgcagcca atatgggatc
     2461 ggccattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     2521 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     2581 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     2641 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     2701 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     2761 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     2821 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     2881 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     2941 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     3001 cggcgatgat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     3061 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     3121 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     3181 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     3241 tgacgagttc ttctgagggg atcaattctc tagagctcgc tgatcagcct cgactgtgcc
     3301 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3361 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3421 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3481 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     3541 ctggggctcg agtgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     3601 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     3661 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3721 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3781 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3841 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3901 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3961 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4021 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4081 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4141 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4201 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4261 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4321 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4381 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4441 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4501 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4561 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4621 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4681 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4741 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4801 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4861 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4921 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4981 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5041 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5101 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5161 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5221 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5281 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5341 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5401 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5461 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5521 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5581 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5641 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5701 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5761 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatat tattgactag
     5821 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     5881 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     5941 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     6001 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     6061 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     6121 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     6181 ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc
     6241 aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg
     6301 ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg cggggcgagg cggagaggtg
     6361 cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc
     6421 ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc
     6481 gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt
     6541 tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg
     6601 tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg
     6661 gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg
     6721 ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc gcggggcttt
     6781 gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg cggtgccccg cggtgcgggg
     6841 ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg
     6901 gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga gttgctgagc
     6961 acggcccggc ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc gccgtgccgg
     7021 gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gcc
//
