LOCUS       sequence-193257-        7943 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses putative OCT4 DBD with a T7 tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7943)
  AUTHORS   Narayan S, Bryant G, Shah S, Berrozpe G, Ptashne M
  TITLE     OCT4 and SOX2 Work as Transcriptional Activators in Reprogramming 
            Human Fibroblasts.
  JOURNAL   Cell Rep. 2017 Aug 15;20(7):1585-1596. doi: 
            10.1016/j.celrep.2017.07.071.
  PUBMED    28813671
REFERENCE   2  (bases 1 to 7943)
  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: 
            "10.1016/j.celrep.2017.07"; journalName: "Cell Rep"; date: 
            "2017-08-15- 15"; volume: "20"; issue: "7"; pages: "1585-1596"
COMMENT     Sequence Label: pSN61
FEATURES             Location/Qualifiers
     source          1..7943
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..511
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     430..448
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             join(526..528,529..531,532..1242)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLFIQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(571..592)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(832..851)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1179..1200
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1252..1308
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     misc_feature    1810..2398
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1863..1883)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2281..2292)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2551..2731
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    2803..2924
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2840..2859)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2894..2913
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2964..3099
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3119..3138)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3120..3138)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3148..3165)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3148..3164)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3157..3179)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3306..3761
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3393..3412)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3603..3624
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3787..3891
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3892..3960,3961..4752)
                     /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     complement(4110..4129)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4923..5511
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5412..5431
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5665..5682
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5799..5820
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5835..5840,5841..5858,5859..5865)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5873..5889
                     /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)."
     primer_bind     5878..5900
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5897..5913
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5897..5913
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5932..5952
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5934..5952
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5980..6206
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6207..6387
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6434..6559
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7052..7285
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7470..7514
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    7781..7898
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ggggttgggg ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc
       61 tgggcgtggt tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc
      121 cgttcgcagc gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc
      181 tgctccgccc ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac
      241 ggaagccgca cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc
      301 gcgccgaccg cgatgggctg tggccaatag cggctgctca gcagggcgcg ccgagagcag
      361 cggccgggaa ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct
      421 gcccgcgcgg tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct
      481 cgttgaccga atcaccgacc tctctcccca gggggatcca ccggtatggt gagcaagggc
      541 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
      601 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
      661 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttc
      721 agctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
      781 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcagcttcaa ggacgacggc
      841 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
      901 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
      961 tacaacagcc acaacgtcta tatcacggcc gacaagcaga agaacggcat caaggcgaac
     1021 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     1081 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gttcatccag
     1141 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     1201 accgccgccg ggatcactca cggcatggac gagctgtaca agggatctgg agcaacaaac
     1261 ttctcactac tcaaacaagc aggtgacgtg gaggagaatc ccggccctat ggacatcaaa
     1321 gctctgcaga aagaactcga gcaatttgcc aagctcctga agcagaagag gatcaccctg
     1381 ggatatacac aggccgatgt ggggctcacc ctgggggttc tatttgggaa ggtattcagc
     1441 caaacgacca tctgccgctt tgaggctctg cagcttagct tcaagaacat gtgtaagctg
     1501 cggcccttgc tgcagaagtg ggtggaggaa gctgacaaca atgaaaatct tcaggagata
     1561 tgcaaagcag aaaccctcgt gcaggcccga aagagaaagc gaaccagtat cgagaaccga
     1621 gtgagaggca acctggagaa tttgttcctg cagtgcccga aacccacact gcagcagatc
     1681 agccacatcg cccagcagct tgggctcgag aaggatgtgg tccgagtgtg gttctgtaac
     1741 cggcgccaga agggcaagcg atcaagcatg gcctccatga ccggcggcca gcagatgggc
     1801 tgagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     1861 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     1921 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     1981 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     2041 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     2101 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     2161 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     2221 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     2281 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     2341 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     2401 ggaattcgag ctcggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     2461 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagata
     2521 tcctgctagc tccggatccg gacctgtact gggtctctct ggttagacca gatctgagcc
     2581 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     2641 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     2701 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag
     2761 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct
     2821 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     2881 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag
     2941 ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3001 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     3061 agctattcca gaagtagtga ggaggctttt ttggaggcct agggacgtac ccaattcgcc
     3121 ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga
     3181 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     3241 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     3301 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3361 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3421 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3481 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3541 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3601 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3661 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3721 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
     3781 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3841 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3901 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3961 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     4021 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     4081 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     4141 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     4201 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     4261 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     4321 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     4381 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     4441 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4501 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4561 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4621 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4681 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4741 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4801 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4861 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4921 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4981 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5041 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     5101 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5161 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5221 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5281 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5341 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5401 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5461 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5521 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5581 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     5641 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     5701 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     5761 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     5821 taggcacccc aggttttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     5881 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac
     5941 cctcactaaa gggaacaaaa gctggagctg caagcttaat gtagtcttat gcaatactct
     6001 tgtagtcttg caacatggta acgatgagtt agcaacatgc cttacaagga gagaaaaagc
     6061 accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac
     6121 agacgggtct gacatggatt ggacgaacca ctgaattgcc gcattgcaga gatattgtat
     6181 ttaagtgcct agctcgatac ataaacgggt ctctctggtt agaccagatc tgagcctggg
     6241 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     6301 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     6361 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
     6421 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
     6481 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
     6541 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
     6601 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
     6661 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
     6721 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
     6781 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
     6841 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
     6901 cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata
     6961 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag
     7021 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag
     7081 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat
     7141 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc
     7201 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa
     7261 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca
     7321 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc
     7381 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
     7441 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
     7501 aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat
     7561 tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag
     7621 tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga
     7681 ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat
     7741 ccattcgatt agtgaacgga tctcgacggt atcggttaac ttttaaaaga aaagggggga
     7801 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
     7861 aagaattaca aaaacaaatt acaaaaattc aaaattttat cgatcacgag actagcctcg
     7921 agaagcttga tatcgaattc cac
//
