LOCUS       sequence-193259-        7703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses putative SOX2 DBD with a V5 tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7703)
  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 7703)
  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: pSN65
FEATURES             Location/Qualifiers
     source          1..7703
                     /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="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     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"
     CDS             1519..1560
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1570..2158
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1623..1643)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2041..2052)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2311..2491
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    2563..2684
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2600..2619)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2654..2673
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2724..2859
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2879..2898)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2880..2898)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2908..2925)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2908..2924)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2917..2939)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3066..3521
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3153..3172)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3363..3384
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3547..3651
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3652..3720,3721..4512)
                     /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(3870..3889)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4683..5271
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5172..5191
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5425..5442
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5559..5580
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5595..5600,5601..5618,5619..5625)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5633..5649
                     /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     5638..5660
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5657..5673
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5657..5673
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5692..5712
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5694..5712
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5740..5966
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5967..6147
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6194..6319
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6812..7045
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7230..7274
                     /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    7541..7658
                     /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 ggctacggcc tgatgtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
      781 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
      841 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
      901 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
      961 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     1021 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     1081 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag
     1141 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     1201 accgccgccg ggatcactct cggcatggac gagctgtaca agggatctgg agcaacaaac
     1261 ttctcactac tcaaacaagc aggtgacgtg gaggagaatc ccggccctat ggtcaagcgg
     1321 cccatgaatg ccttcatggt gtggtcccgc gggcagcggc gcaagatggc ccaggagaac
     1381 cccaagatgc acaactcgga gatcagcaag cgcctgggcg ccgagtggaa acttttgtcg
     1441 gagacggaga agcggccgtt catcgacgag gctaagcggc tgcgagcgct gcacatgaag
     1501 gagcacccgg attataaagg caagcccatc cccaaccccc tgctgggcct ggactccacc
     1561 tgagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     1621 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     1681 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     1741 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     1801 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     1861 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     1921 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     1981 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     2041 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     2101 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     2161 ggaattcgag ctcggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     2221 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagata
     2281 tcctgctagc tccggatccg gacctgtact gggtctctct ggttagacca gatctgagcc
     2341 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     2401 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     2461 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag
     2521 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct
     2581 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     2641 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag
     2701 ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     2761 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     2821 agctattcca gaagtagtga ggaggctttt ttggaggcct agggacgtac ccaattcgcc
     2881 ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga
     2941 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     3001 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     3061 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3121 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3181 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3241 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3301 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3361 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3421 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3481 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
     3541 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3601 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3661 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3721 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3781 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3841 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     3901 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3961 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     4021 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     4081 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     4141 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     4201 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4261 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4321 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4381 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4441 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4501 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4561 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4621 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4681 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4741 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4801 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4861 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4921 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4981 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5041 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5101 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5161 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5221 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5281 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5341 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     5401 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     5461 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     5521 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     5581 taggcacccc aggttttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     5641 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac
     5701 cctcactaaa gggaacaaaa gctggagctg caagcttaat gtagtcttat gcaatactct
     5761 tgtagtcttg caacatggta acgatgagtt agcaacatgc cttacaagga gagaaaaagc
     5821 accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac
     5881 agacgggtct gacatggatt ggacgaacca ctgaattgcc gcattgcaga gatattgtat
     5941 ttaagtgcct agctcgatac ataaacgggt ctctctggtt agaccagatc tgagcctggg
     6001 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     6061 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     6121 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
     6181 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
     6241 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
     6301 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
     6361 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
     6421 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
     6481 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
     6541 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
     6601 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
     6661 cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata
     6721 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag
     6781 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag
     6841 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat
     6901 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc
     6961 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa
     7021 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca
     7081 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc
     7141 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
     7201 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
     7261 aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat
     7321 tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag
     7381 tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga
     7441 ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat
     7501 ccattcgatt agtgaacgga tctcgacggt atcggttaac ttttaaaaga aaagggggga
     7561 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
     7621 aagaattaca aaaacaaatt acaaaaattc aaaattttat cgatcacgag actagcctcg
     7681 agaagcttga tatcgaattc cac
//
