LOCUS       sequence-222650-        8506 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of an OCT4 (Pou5f1) protein tagged with 
            SNAP-tag and a non-functional mitotic degron (MDMUT) for control.
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-MDMUT-OCT4
FEATURES             Location/Qualifiers
     source          1..8506
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    49..70
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(85..90,91..108,109..115)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    123..139
                     /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     128..150
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     147..163
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     147..163
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     182..202
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        184..202
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        230..456
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             457..637
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    684..809
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1302..1535
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1720..1764
                     /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    2031..2148
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2194..2704
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2623..2641
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2719..3264
                     /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    4579..5167
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4632..4652)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5050..5061)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5254..5487
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5559..5680
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5596..5615)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5650..5669
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5720..5855
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5875..5894)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5876..5894)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5904..5921)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5904..5920)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5913..5935)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6062..6517
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6149..6168)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6359..6380
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6543..6647
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6648..6716,6717..7508)
                     /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(6866..6885)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7679..8267
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8168..8187
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8421..8438
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
       61 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
      121 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg
      181 cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttaat gtagtcttat
      241 gcaatactct tgtagtcttg caacatggta acgatgagtt agcaacatgc cttacaagga
      301 gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg tgccttatta
      361 ggaaggcaac agacgggtct gacatggatt ggacgaacca ctgaattgcc gcattgcaga
      421 gatattgtat ttaagtgcct agctcgatac ataaacgggt ctctctggtt agaccagatc
      481 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
      541 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
      601 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
      661 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
      721 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
      781 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
      841 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
      901 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
      961 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
     1021 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
     1081 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
     1141 agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
     1201 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     1261 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     1321 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     1381 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     1441 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     1501 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     1561 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     1621 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     1681 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     1741 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     1801 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     1861 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     1921 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     1981 agagacagat ccattcgatt agtgaacgga tctcgacggt atcggttaac ttttaaaaga
     2041 aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac
     2101 atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttat cgatcacgag
     2161 actagcctcg agaagcttga tatcgaattc cacggggttg gggttgcgcc ttttccaagg
     2221 cagccctggg tttgcgcagg gacgcggctg ctctgggcgt ggttccggga aacgcagcgg
     2281 cgccgaccct gggtctcgca cattcttcac gtccgttcgc agcgtcaccc ggatcttcgc
     2341 cgctaccctt gtgggccccc cggcgacgct tcctgctccg cccctaagtc gggaaggttc
     2401 cttgcggttc gcggcgtgcc ggacgtgaca aacggaagcc gcacgtctca ctagtaccct
     2461 cgcagacgga cagcgccagg gagcaatggc agcgcgccga ccgcgatggg ctgtggccaa
     2521 tagcggctgc tcagcagggc gcgccgagag cagcggccgg gaaggggcgg tgcgggaggc
     2581 ggggtgtggg gcggtagtgt gggccctgtt cctgcccgcg cggtgttccg cattctgcaa
     2641 gcctccggag cgcacgtcgg cagtcggctc cctcgttgac cgaatcaccg acctctctcc
     2701 ccagggggat ccaccggtat ggacaaagac tgcgaaatga agcgcaccac cctggatagc
     2761 cctctgggca agctggaact gtctgggtgc gaacagggcc tgcaccgtat catcttcctg
     2821 ggcaaaggaa catctgccgc cgacgccgtg gaagtgcctg ccccagccgc cgtgctgggc
     2881 ggaccagagc cactgatgca ggccaccgcc tggctcaacg cctactttca ccagcctgag
     2941 gccatcgagg agttccctgt gccagccctg caccacccag tgttccagca ggagagcttt
     3001 acccgccagg tgctgtggaa actgctgaaa gtggtgaagt tcggagaggt catcagctac
     3061 agccacctgg ccgccctggc cggcaatccc gccgccaccg ccgccgtgaa aaccgccctg
     3121 agcggaaatc ccgtgcccat tctgatcccc tgccaccggg tggtgcaggg cgacctggac
     3181 gtggggggct acgagggcgg gctcgccgtg aaagagtggc tgctggccca cgagggccac
     3241 agactgggca agcctgggct gggttctaga gcagaaaata aggctaaagt ctcaatggcc
     3301 ggcgctaagc gcgttcccgt gaccgtaact gccgcttcta aacccgggtt gcgcccggcg
     3361 actgctcttg gagatattgg caacaaggtg tctgaagagc tgcaggctcg cgtgccactg
     3421 aagcgagagg ctaagacgct gggaactgga aagggtacgg ttaaggccct gccgaagcca
     3481 gtggagaaag ttcctgtctg cgaaccggct agaatggctg gacacctggc ttcagacttc
     3541 gccttctcac ccccaccagg tgggggtgat gggtcagcag ggctggagcc gggctgggtg
     3601 gatcctcgaa cctggctaag cttccaaggg cctccaggtg ggcctggaat cggaccaggc
     3661 tcagaggtat tggggatctc cccatgtccg cccgcatacg agttctgcgg agggatggca
     3721 tactgtggac ctcaggttgg actgggccta gtcccccaag ttggcgtgga gactttgcag
     3781 cctgagggcc aggcaggagc acgagtggaa agcaactcag agggaacctc ctctgagccc
     3841 tgtgccgacc gccccaatgc cgtgaagttg gagaaggtgg aaccaactcc cgaggagtcc
     3901 caggacatga aagccctgca gaaggagcta gaacagtttg ccaagctgct gaagcagaag
     3961 aggatcacct tggggtacac ccaggccgac gtggggctca ccctgggcgt tctctttgga
     4021 aaggtgttca gccagaccac catctgtcgc ttcgaggcct tgcagctcag ccttaagaac
     4081 atgtgtaagc tgcggcccct gctggagaag tgggtggagg aagccgacaa caatgagaac
     4141 cttcaggaga tatgcaaatc ggagaccctg gtgcaggccc ggaagagaaa gcgaactagc
     4201 attgagaacc gtgtgaggtg gagtctggag accatgtttc tgaagtgccc gaagccctcc
     4261 ctacagcaga tcactcacat cgccaatcag cttgggctag agaaggatgt ggttcgagta
     4321 tggttctgta accggcgcca gaagggcaaa agatcaagta ttgagtattc ccaacgagaa
     4381 gagtatgagg ctacagggac acctttccca gggggggctg tatcctttcc tctgccccca
     4441 ggtccccact ttggcacccc aggctatgga agcccccact tcaccacact ctactcagtc
     4501 ccttttcctg agggcgaggc ctttccctct gttcccgtca ctgctctggg ctctcccatg
     4561 cattcaaact gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     4621 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     4681 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     4741 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     4801 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     4861 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     4921 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
     4981 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     5041 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     5101 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     5161 tccccgcctg gaattcgagc tcggtacctt taagaccaat gacttacaag gcagctgtag
     5221 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     5281 gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     5341 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     5401 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     5461 tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt
     5521 tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata
     5581 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5641 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat
     5701 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     5761 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     5821 attccagaag tagtgaggag gcttttttgg aggcctaggg acgtacccaa ttcgccctat
     5881 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     5941 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     6001 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     6061 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     6121 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     6181 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     6241 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     6301 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     6361 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     6421 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     6481 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
     6541 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     6601 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     6661 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     6721 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     6781 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     6841 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     6901 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     6961 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     7021 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     7081 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     7141 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     7201 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     7261 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     7321 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     7381 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     7441 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     7501 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     7561 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     7621 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     7681 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     7741 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     7801 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     7861 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     7921 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     7981 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     8041 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     8101 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     8161 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     8221 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     8281 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     8341 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     8401 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     8461 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctg
//
