LOCUS       sequence-401144-        6855 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Competitive assay of Oct4 and Oct6 binding to DNA motifs..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6855)
  AUTHORS   Mistri TK, Devasia AG, Chu LT, Ng WP, Halbritter F, Colby D, 
            Martynoga B, Tomlinson SR, Chambers I, Robson P, Wohland T
  TITLE     Selective influence of Sox2 on POU transcription factor binding in 
            embryonic and neural stem cells.
  JOURNAL   EMBO Rep. 2015 Sep;16(9):1177-91. doi: 10.15252/embr.201540467. Epub
            2015 Aug 11.
  PUBMED    26265007
REFERENCE   2  (bases 1 to 6855)
  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: 
            "10.15252/embr.201540467"; journalName: "EMBO Rep"; date: "2015-09";
            volume: "16"; issue: "9"; pages: "1177-91"
COMMENT     Sequence Label: pCAG eGFP-Oct4
FEATURES             Location/Qualifiers
     source          1..6855
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          644..1660
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1584..1606
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1668..1687
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(1733..1735,1736..1738,1739..2449)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1778..1799)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2039..2058)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2386..2407
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3643..3764
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3680..3699)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3734..3753
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3771..4226)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3908..3929)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4120..4139
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4253..4357
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4359..4716
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4359..4550
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4382..4402)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4567..4702
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4751..5545
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4751..5545
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4805..4824)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5415..5434
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5733..5752)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5777..5824
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6153..6741
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6642..6661
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc
      421 cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg
      481 gggggggcgc gcgccrggsg gggsggggsg gggsgggggg sggggsgggg sgrggsggrg
      541 rggkgcggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg
      601 gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg
      661 ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac
      721 cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg
      781 cttggtttaa tgacggcttg tttcttttct gtggctgcgt gaaagccttg aggggctccg
      841 ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg
      901 gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg
      961 gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg
     1021 cggggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc
     1081 agggggtgtg ggcgcgtcgg tcgggctgca accccccctg cacccccctc cccgagttgc
     1141 tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt
     1201 gccgggcggg gggtggcggc aggtgggggt gccgggcggg gcggggccgc ctcgggccgg
     1261 ggrgggctcg ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc
     1321 gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc
     1381 ccaaatctgt gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg
     1441 ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc
     1501 gcgccgccgt ccccttctcc ctctccagcc tcggggctgt ccgcgggggg acggctgcct
     1561 tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gctctagagc
     1621 ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt
     1681 tattgtgctg tctcatcatt ttggcaaaga attgagctct atggccacaa ccatggtgag
     1741 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     1801 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     1861 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     1921 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     1981 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     2041 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     2101 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     2161 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     2221 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     2281 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     2341 cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     2401 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg gcggcagcgg
     2461 catggctgga cacctggctt cagacttcgc cttctcaccc ccaccaggtg ggggtgatgg
     2521 gtcagcaggg ctggagccgg gctgggtgga tcctcgaacc tggctaagct tccaagggcc
     2581 tccaggtggg cctggaatcg gaccaggctc agaggtattg gggatctccc catgtccgcc
     2641 cgcatacgag ttctgcggag ggatggcata ctgtggacct caggttggac tgggcctagt
     2701 cccccaagtt ggcgtggaga ctttgcagcc tgagggccag gcaggagcac gagtggaaag
     2761 caactcagag ggaacctcct ctgagccctg tgccgaccgc cccaatgccg tgaagttgga
     2821 gaaggtggaa ccaactcccg aggagtccca ggacatgaaa gccctgcaga aggagctaga
     2881 acagtttgcc aagctgctga agcagaagag gatcaccttg gggtacaccc aggccgacgt
     2941 ggggctcacc ctgggcgttc tctttggaaa ggtgttcagc cagaccacca tctgtcgctt
     3001 cgaggccttg cagctcagcc ttaagaacat gtgtaagctg cggcccctgc tggagaagtg
     3061 ggtggaggaa gccgacaaca atgagaacct tcaggagata tgcaaatcgg agaccctggt
     3121 gcaggcccgg aagagaaagc gaactagcat tgagaaccgt gtgaggtgga gtctggagac
     3181 catgtttctg aagtgcccga agccctccct acagcagatc actcacatcg ccaatcagct
     3241 tgggctagag aaggatgtgg ttcgagtatg gttctgtaac cggcgccaga agggcaaaag
     3301 atcaagtatt gagtattccc aacgagaaga gtatgaggct acagggacac ctttcccagg
     3361 gggggctgta tcctttcctc tgcccccagg tccccacttt ggcaccccag gctatggaag
     3421 cccccacttc accacactct actcagtccc ttttcctgag ggcgaggcct ttccctctgt
     3481 tcccgtcact gctctgggct ctcccatgca ttcaaactga agcggccgcg actctagatc
     3541 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     3601 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     3661 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     3721 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag
     3781 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     3841 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     3901 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     3961 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     4021 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     4081 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     4141 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     4201 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     4261 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4321 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     4381 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4441 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4501 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4561 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4621 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     4681 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     4741 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4801 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4861 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4921 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4981 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     5041 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     5101 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     5161 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     5221 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     5281 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     5341 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     5401 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     5461 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     5521 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     5581 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     5641 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     5701 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     5761 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     5821 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     5881 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     5941 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     6001 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6061 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6121 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6181 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6241 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6301 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6361 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6421 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6481 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6541 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6601 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6661 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6721 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6781 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     6841 attaccgcca tgcat
//
