LOCUS       sequence-218576-        8502 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of an OCT4 (Pou5f1) protein tagged with 
            SNAP-tag and a mitotic degron (MD).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8502)
  AUTHORS   Friman ET, Deluz C, Meireles-Filho AC, Govindan S, Gardeux V, 
            Deplancke B, Suter DM
  TITLE     Dynamic regulation of chromatin accessibility by pluripotency 
            transcription factors across the cell cycle.
  JOURNAL   eLife. 2019 Dec 3;8. pii: 50087. doi: 10.7554/eLife.50087.
  PUBMED    31794382
REFERENCE   2  (bases 1 to 8502)
  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: "eLife."; 
            date: "2019-12-3"; volume: "8. pii: 50087. doi"; pages: " 
            10.7554/eLife.50087"
COMMENT     Sequence Label: pLV-PGK-SNAP-MD-OCT4
FEATURES             Location/Qualifiers
     source          1..8502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             236..416
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    460..585
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1078..1311
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1496..1540
                     /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    1807..1924
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1971..2481
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2400..2418
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2496..3041
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     misc_feature    4356..4944
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4409..4429)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4827..4838)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5031..5264
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5336..5457
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5373..5392)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5427..5446
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5476..5611
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5643..5662)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5644..5662)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5672..5689)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5672..5688)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5681..5703)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5833..6288
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5920..5939)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6130..6151
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6317..6421
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6422..6490,6491..7282)
                     /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(6640..6659)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7453..8041
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7942..7961
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8195..8212
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8329..8350
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8365..8370,8371..8388,8389..8395)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8403..8419
                     /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     8408..8430
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8427..8443
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8427..8443
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8462..8482
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8464..8482
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
       61 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
      121 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
      181 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacaa taaacgggtc
      241 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
      301 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
      361 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
      421 cgcccgaaca gggacctgaa agcgaaaggg aaaccagagc tctctcgacg caggactcgg
      481 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt
      541 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg
      601 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa
      661 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg
      721 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca
      781 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa
      841 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa
      901 agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag gagatatgag
      961 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
     1021 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
     1081 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cctcaatgac
     1141 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
     1201 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
     1261 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
     1321 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
     1381 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
     1441 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
     1501 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
     1561 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
     1621 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
     1681 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
     1741 aggtggagag agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg
     1801 gttaactttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
     1861 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa
     1921 ttttatcgat cacgagacta gcctcgagaa gcttgatatc gaattcccac ggggttgggg
     1981 ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc tgggcgtggt
     2041 tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc
     2101 gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc tgctccgccc
     2161 ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca
     2221 cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc gcgccgaccg
     2281 cgatgggctg tggccaatag cggctgctca gcggggcgcg ccgagagcag cggccgggaa
     2341 ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg
     2401 tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct cgttgaccga
     2461 atcaccgacc tctctcccca gggggatcca ccggtatgga caaagactgc gaaatgaagc
     2521 gcaccaccct ggatagccct ctgggcaagc tggaactgtc tgggtgcgaa cagggcctgc
     2581 accgtatcat cttcctgggc aaaggaacat ctgccgccga cgccgtggaa gtgcctgccc
     2641 cagccgccgt gctgggcgga ccagagccac tgatgcaggc caccgcctgg ctcaacgcct
     2701 actttcacca gcctgaggcc atcgaggagt tccctgtgcc agccctgcac cacccagtgt
     2761 tccagcagga gagctttacc cgccaggtgc tgtggaaact gctgaaagtg gtgaagttcg
     2821 gagaggtcat cagctacagc cacctggccg ccctggccgg caatcccgcc gccaccgccg
     2881 ccgtgaaaac cgccctgagc ggaaatcccg tgcccattct gatcccctgc caccgggtgg
     2941 tgcagggcga cctggacgtg gggggctacg agggcgggct cgccgtgaaa gagtggctgc
     3001 tggcccacga gggccacaga ctgggcaagc ctgggctggg ttctagagca gaaaataagg
     3061 ctaaagtctc aatggccggc gctaagcgcg ttcccgtgac cgtaactgcc gcttctaaac
     3121 ccgggttgcg cccgaggact gctcttggag atattggcaa caaggtgtct gaagagctgc
     3181 aggctcgcgt gccactgaag cgagaggcta agacgctggg aactggaaag ggtacggtta
     3241 aggccctgcc gaagccagtg gagaaagttc ctgtctgcga accggctaga atggctggac
     3301 acctggcttc agacttcgcc ttctcacccc caccaggtgg gggtgatggg tcagcagggc
     3361 tggagccggg ctgggtggat cctcgaacct ggctaagctt ccaagggcct ccaggtgggc
     3421 ctggaatcgg accaggctca gaggtattgg ggatctcccc atgtccgccc gcatacgagt
     3481 tctgcggagg gatggcatac tgtggacctc aggttggact gggcctagtc ccccaagttg
     3541 gcgtggagac tttgcagcct gagggccagg caggagcacg agtggaaagc aactcagagg
     3601 gaacctcctc tgagccctgt gccgaccgcc ccaatgccgt gaagttggag aaggtggaac
     3661 caactcccga ggagtcccag gacatgaaag ccctgcagaa ggagctagaa cagtttgcca
     3721 agctgctgaa gcagaagagg atcaccttgg ggtacaccca ggccgacgtg gggctcaccc
     3781 tgggcgttct ctttggaaag gtgttcagcc agaccaccat ctgtcgcttc gaggccttgc
     3841 agctcagcct taagaacatg tgtaagctgc ggcccctgct ggagaagtgg gtggaggaag
     3901 ccgacaacaa tgagaacctt caggagatat gcaaatcgga gaccctggtg caggcccgga
     3961 agagaaagcg aactagcatt gagaaccgtg tgaggtggag tctggagacc atgtttctga
     4021 agtgcccgaa gccctcccta cagcagatca ctcacatcgc caatcagctt gggctagaga
     4081 aggatgtggt tcgagtatgg ttctgtaacc ggcgccagaa gggcaaaaga tcaagtattg
     4141 agtattccca acgagaagag tatgaggcta cagggacacc tttcccaggg ggggctgtat
     4201 cctttcctct gcccccaggt ccccactttg gcaccccagg ctatggaagc ccccacttca
     4261 ccacactcta ctcagtccct tttcctgagg gcgaggcctt tccctctgtt cccgtcactg
     4321 ctctgggctc tcccatgcat tcaaactgag tcgacaatca acctctggat tacaaaattt
     4381 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     4441 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     4501 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     4561 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     4621 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     4681 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     4741 tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc
     4801 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     4861 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     4921 tctccctttg ggccgcctcc ccgcctggaa ttcgagctcg gtacctttaa gaccaatgac
     4981 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     5041 aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga
     5101 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     5161 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     5221 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca
     5281 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt
     5341 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5401 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5461 tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     5521 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     5581 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcgtcgagac gtacccaatt
     5641 cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact
     5701 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     5761 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     5821 gcgaatggcg cgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     5881 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     5941 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     6001 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     6061 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     6121 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     6181 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     6241 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttcc caggtggcac
     6301 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     6361 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     6421 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     6481 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     6541 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     6601 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     6661 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     6721 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     6781 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     6841 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     6901 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     6961 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     7021 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     7081 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     7141 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     7201 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     7261 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     7321 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     7381 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     7441 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     7501 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     7561 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     7621 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     7681 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     7741 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     7801 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     7861 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     7921 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     7981 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     8041 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     8101 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     8161 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     8221 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     8281 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     8341 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     8401 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg
     8461 cgcaattaac cctcactaaa gggaacaaaa gctggagctg ca
//
