LOCUS       sequence-222649-        8506 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 8506)
  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 8506)
  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..8506
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    102..123
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(138..143,144..161,162..168)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    176..192
                     /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     181..203
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     200..216
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     200..216
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     235..255
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        237..255
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        283..509
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             510..690
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    737..862
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1355..1588
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1773..1817
                     /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    2084..2201
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2247..2757
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2676..2694
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2772..3317
                     /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    4632..5220
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4685..4705)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5103..5114)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5307..5540
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5612..5733
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5649..5668)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5703..5722
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5773..5908
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5928..5947)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5929..5947)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5957..5974)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5957..5973)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5966..5988)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6115..6570
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6202..6221)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6412..6433
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6596..6700
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6701..6769,6770..7561)
                     /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(6919..6938)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7732..8320
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8221..8240
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8474..8491
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
       61 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
      121 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
      181 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
      241 aaccctcact aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac
      301 tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa
      361 agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc
      421 aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg
      481 tatttaagtg cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct
      541 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
      601 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
      661 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
      721 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
      781 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
      841 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
      901 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
      961 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     1021 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     1081 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     1141 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     1201 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     1261 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     1321 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     1381 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     1441 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     1501 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     1561 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     1621 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     1681 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     1741 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     1801 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     1861 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
     1921 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
     1981 cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca
     2041 gatccattcg attagtgaac ggatctcgac ggtatcggtt aacttttaaa agaaaagggg
     2101 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     2161 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tatcgatcac gagactagcc
     2221 tcgagaagct tgatatcgaa ttccacgggg ttggggttgc gccttttcca aggcagccct
     2281 gggtttgcgc agggacgcgg ctgctctggg cgtggttccg ggaaacgcag cggcgccgac
     2341 cctgggtctc gcacattctt cacgtccgtt cgcagcgtca cccggatctt cgccgctacc
     2401 cttgtgggcc ccccggcgac gcttcctgct ccgcccctaa gtcgggaagg ttccttgcgg
     2461 ttcgcggcgt gccggacgtg acaaacggaa gccgcacgtc tcactagtac cctcgcagac
     2521 ggacagcgcc agggagcaat ggcagcgcgc cgaccgcgat gggctgtggc caatagcggc
     2581 tgctcagcag ggcgcgccga gagcagcggc cgggaagggg cggtgcggga ggcggggtgt
     2641 ggggcggtag tgtgggccct gttcctgccc gcgcggtgtt ccgcattctg caagcctccg
     2701 gagcgcacgt cggcagtcgg ctccctcgtt gaccgaatca ccgacctctc tccccagggg
     2761 gatccaccgg tatggacaaa gactgcgaaa tgaagcgcac caccctggat agccctctgg
     2821 gcaagctgga actgtctggg tgcgaacagg gcctgcaccg tatcatcttc ctgggcaaag
     2881 gaacatctgc cgccgacgcc gtggaagtgc ctgccccagc cgccgtgctg ggcggaccag
     2941 agccactgat gcaggccacc gcctggctca acgcctactt tcaccagcct gaggccatcg
     3001 aggagttccc tgtgccagcc ctgcaccacc cagtgttcca gcaggagagc tttacccgcc
     3061 aggtgctgtg gaaactgctg aaagtggtga agttcggaga ggtcatcagc tacagccacc
     3121 tggccgccct ggccggcaat cccgccgcca ccgccgccgt gaaaaccgcc ctgagcggaa
     3181 atcccgtgcc cattctgatc ccctgccacc gggtggtgca gggcgacctg gacgtggggg
     3241 gctacgaggg cgggctcgcc gtgaaagagt ggctgctggc ccacgagggc cacagactgg
     3301 gcaagcctgg gctgggttct agagcagaaa ataaggctaa agtctcaatg gccggcgcta
     3361 agcgcgttcc cgtgaccgta actgccgctt ctaaacccgg gttgcgcccg aggactgctc
     3421 ttggagatat tggcaacaag gtgtctgaag agctgcaggc tcgcgtgcca ctgaagcgag
     3481 aggctaagac gctgggaact ggaaagggta cggttaaggc cctgccgaag ccagtggaga
     3541 aagttcctgt ctgcgaaccg gctagaatgg ctggacacct ggcttcagac ttcgccttct
     3601 cacccccacc aggtgggggt gatgggtcag cagggctgga gccgggctgg gtggatcctc
     3661 gaacctggct aagcttccaa gggcctccag gtgggcctgg aatcggacca ggctcagagg
     3721 tattggggat ctccccatgt ccgcccgcat acgagttctg cggagggatg gcatactgtg
     3781 gacctcaggt tggactgggc ctagtccccc aagttggcgt ggagactttg cagcctgagg
     3841 gccaggcagg agcacgagtg gaaagcaact cagagggaac ctcctctgag ccctgtgccg
     3901 accgccccaa tgccgtgaag ttggagaagg tggaaccaac tcccgaggag tcccaggaca
     3961 tgaaagccct gcagaaggag ctagaacagt ttgccaagct gctgaagcag aagaggatca
     4021 ccttggggta cacccaggcc gacgtggggc tcaccctggg cgttctcttt ggaaaggtgt
     4081 tcagccagac caccatctgt cgcttcgagg ccttgcagct cagccttaag aacatgtgta
     4141 agctgcggcc cctgctggag aagtgggtgg aggaagccga caacaatgag aaccttcagg
     4201 agatatgcaa atcggagacc ctggtgcagg cccggaagag aaagcgaact agcattgaga
     4261 accgtgtgag gtggagtctg gagaccatgt ttctgaagtg cccgaagccc tccctacagc
     4321 agatcactca catcgccaat cagcttgggc tagagaagga tgtggttcga gtatggttct
     4381 gtaaccggcg ccagaagggc aaaagatcaa gtattgagta ttcccaacga gaagagtatg
     4441 aggctacagg gacacctttc ccaggggggg ctgtatcctt tcctctgccc ccaggtcccc
     4501 actttggcac cccaggctat ggaagccccc acttcaccac actctactca gtcccttttc
     4561 ctgagggcga ggcctttccc tctgttcccg tcactgctct gggctctccc atgcattcaa
     4621 actgagtcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     4681 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     4741 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     4801 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     4861 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     4921 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     4981 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     5041 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     5101 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     5161 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     5221 ctggaattcg agctcggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
     5281 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
     5341 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     5401 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     5461 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     5521 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact
     5581 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta
     5641 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5701 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc
     5761 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     5821 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     5881 aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt
     5941 cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     6001 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     6061 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
     6121 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     6181 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     6241 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     6301 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     6361 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     6421 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     6481 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     6541 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     6601 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     6661 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     6721 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     6781 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     6841 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     6901 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     6961 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     7021 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     7081 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     7141 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     7201 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     7261 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     7321 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     7381 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     7441 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     7501 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     7561 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     7621 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     7681 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     7741 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     7801 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     7861 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     7921 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     7981 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     8041 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     8101 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     8161 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     8221 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     8281 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     8341 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     8401 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     8461 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgc
//
