LOCUS       sequence-401148-        7201 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 7201)
  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 7201)
  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-Oct6
FEATURES             Location/Qualifiers
     source          1..7201
                     /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(1727..1729,1730..1732,1733..2443)
                     /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(1772..1793)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2033..2052)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2380..2401
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3770..3787
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3989..4110
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4026..4045)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4080..4099
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4117..4572)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4254..4275)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4466..4485
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4599..4703
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4705..5062
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4705..4896
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4728..4748)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4913..5048
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5097..5891
                     /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             5097..5891
                     /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(5151..5170)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5761..5780
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6079..6098)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6123..6170
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6499..7087
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6988..7007
                     /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 gggsgggggg 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 aggkgggggt gccgggcggg gcggggccgc ctcgggccgg
     1261 ggrgggctcg ggggrggggc 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 attgagctcc acaaccatgg tgagcaaggg
     1741 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     1801 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     1861 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     1921 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     1981 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     2041 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     2101 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     2161 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     2221 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     2281 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     2341 gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     2401 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggcggca gcggcatggc
     2461 caccaccgcg cagtatctgc cgcggggccc cggcggcgga gctgggggca cagggccgct
     2521 catgcatccc gatgccgccg cggcggcggc agcggcggcg gcggccgagc ggctgcacgc
     2581 gggggccgcg taccgcgaag tgcagaagct gatgcaccac gagtggctgg gcgcgggcgc
     2641 gggccacccc gtgggcctag cgcaccctca atggctaccc acgggaggag gcggcggcgg
     2701 cgactgggcg ggcggcccgc acctggaaca cggcaaggca ggcggtggcg gtaccggccg
     2761 agctgacgac ggcggcggtg gcggcggttt ccacgcccgc ctggtgcacc aaggggcggc
     2821 ccacgcgggc gcggcatggg cacaaggcgg cacagcgcac cacttgggcc ccgccatgtc
     2881 gccgtcgccc ggggccggcg ggggtcacca gccccagccg ctcgggctgt acgctcaggc
     2941 ggcctacccc ggtggcggcg gcggcggcct ggccgggatg ctggcggcgg gaggcggcgg
     3001 cgcgggaccc ggcctgcacc acgcactgca cgaggacggc cacgaggcac agctggagcc
     3061 gtcgccacca ccgcacctgg gcgcacacgg acacgcacac ggacatgcac acgcgggcgg
     3121 cctgcacgcg gcggcggcgc acctgcaccc gggcgcgggc ggtggtggct cgtcggtggg
     3181 cgagcactcg gacgaggatg ctcccagctc cgacgacctg gagcagttcg ccaagcagtt
     3241 caagcaacga cgcatcaagc tgggcttcac ccaggccgac gtgggactgg cgctgggcac
     3301 cctctacggt aacgtgttct cgcagaccac catctgccgt ttcgaggccc tgcagctgag
     3361 cttcaagaac atgtgcaagc tcaagccgct gctcaacaag tggctggagg agaccgactc
     3421 gtccagcggc agccccacca acctggacaa gatcgcggcg cagggccgca agcgcaagaa
     3481 gcgcacgtcc atcgaggtgg gtgtcaaagg cgcgctcgag agccactttc tcaagtgtcc
     3541 caagccgtct gcgcacgaga tcaccggcct ggccgacagc ctgcaactgg agaaggaggt
     3601 ggtgcgtgtc tggttctgca accggcggca gaaggagaag cgcatgaccc ccgcggccgg
     3661 cgcgggccac ccgcccatgg acgacgttta tgcgcctggg gagctggggc ctggcggggg
     3721 cagcgcgtca ccaccttctg cgcccccgcc acccccgccg gccgcgctgc accaccacca
     3781 ccaccacaca ctgcccggct ctgtgcagtg accctgcgga ctgggttccc cgccggcgca
     3841 gcggtgcctc cggcgcgcag ttagacggcg gccgcgactc tagatcataa tcagccatac
     3901 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     3961 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     4021 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4081 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt
     4141 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     4201 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     4261 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     4321 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     4381 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     4441 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     4501 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     4561 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4621 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4681 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     4741 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4801 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4861 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4921 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4981 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     5041 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     5101 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     5161 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     5221 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     5281 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     5341 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     5401 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     5461 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     5521 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     5581 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     5641 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     5701 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     5761 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     5821 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     5881 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     5941 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     6001 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     6061 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     6121 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     6181 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     6241 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     6301 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     6361 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6421 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6481 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6541 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6601 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     6661 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6721 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6781 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6841 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6901 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6961 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     7021 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     7081 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     7141 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     7201 t
//
