LOCUS       sequence-222587-        8407 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of a SOX2 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 8407)
  AUTHORS   Deluz C, Friman ET, Strebinger D, Benke A, Raccaud M, Callegari A, 
            Leleu M, Manley S, Suter DM
  TITLE     A role for mitotic bookmarking of SOX2 in pluripotency and 
            differentiation.
  JOURNAL   Genes Dev. 2016 Nov 15;30(22):2538-2550. doi: 
            10.1101/gad.289256.116. Epub 2016 Dec 5.
  PUBMED    27920086
REFERENCE   2  (bases 1 to 8407)
  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: 
            "10.1101/gad.289256.116"; journalName: "Genes Dev"; date: 
            "2016-11-15- 15"; volume: "30"; issue: "22"; pages: "2538-2550"
COMMENT     Sequence Label: pLV-PGK-SNAP-MDMUT-SOX2
FEATURES             Location/Qualifiers
     source          1..8407
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    88..109
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(124..129,130..147,148..154)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    162..178
                     /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     167..189
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     186..202
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     186..202
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     221..241
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        223..241
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        269..495
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             496..676
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    723..848
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1341..1574
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1759..1803
                     /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    2070..2187
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2233..2743
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2662..2680
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2758..3303
                     /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    4519..5107
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4572..4592)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4990..5001)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5194..5427
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5499..5620
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5536..5555)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5590..5609
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5660..5795
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5815..5834)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5816..5834)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5844..5861)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5844..5860)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5853..5875)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6002..6457
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6089..6108)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6299..6320
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6483..6587
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6588..6656,6657..7448)
                     /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(6806..6825)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7619..8207
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8108..8127
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8361..8378
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
       61 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
      121 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
      181 tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag
      241 ggaacaaaag ctggagctgc aagcttaatg tagtcttatg caatactctt gtagtcttgc
      301 aacatggtaa cgatgagtta gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc
      361 cgattggtgg aagtaaggtg gtacgatcgt gccttattag gaaggcaaca gacgggtctg
      421 acatggattg gacgaaccac tgaattgccg cattgcagag atattgtatt taagtgccta
      481 gctcgataca taaacgggtc tctctggtta gaccagatct gagcctggga gctctctggc
      541 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
      601 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
      661 tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg
      721 agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc
      781 gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg
      841 agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc
      901 cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac
      961 gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac
     1021 agctacaacc atcccttcag acaggatcag aagaacttag atcattatat aatacagtag
     1081 caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca
     1141 agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc
     1201 agacctggag gaggagatat gagggacaat tggagaagtg aattatataa atataaagta
     1261 gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga
     1321 gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc
     1381 actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata
     1441 gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc
     1501 acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag
     1561 gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg
     1621 ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg
     1681 atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa
     1741 tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt
     1801 ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata
     1861 gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt
     1921 aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac
     1981 aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta
     2041 gtgaacggat ctcgacggta tcggttaact tttaaaagaa aaggggggat tggggggtac
     2101 agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa
     2161 aaacaaatta caaaaattca aaattttatc gatcacgaga ctagcctcga gaagcttgat
     2221 atcgaattcc acggggttgg ggttgcgcct tttccaaggc agccctgggt ttgcgcaggg
     2281 acgcggctgc tctgggcgtg gttccgggaa acgcagcggc gccgaccctg ggtctcgcac
     2341 attcttcacg tccgttcgca gcgtcacccg gatcttcgcc gctacccttg tgggcccccc
     2401 ggcgacgctt cctgctccgc ccctaagtcg ggaaggttcc ttgcggttcg cggcgtgccg
     2461 gacgtgacaa acggaagccg cacgtctcac tagtaccctc gcagacggac agcgccaggg
     2521 agcaatggca gcgcgccgac cgcgatgggc tgtggccaat agcggctgct cagcagggcg
     2581 cgccgagagc agcggccggg aaggggcggt gcgggaggcg gggtgtgggg cggtagtgtg
     2641 ggccctgttc ctgcccgcgc ggtgttccgc attctgcaag cctccggagc gcacgtcggc
     2701 agtcggctcc ctcgttgacc gaatcaccga cctctctccc cagggggatc caccggtatg
     2761 gacaaagact gcgaaatgaa gcgcaccacc ctggatagcc ctctgggcaa gctggaactg
     2821 tctgggtgcg aacagggcct gcaccgtatc atcttcctgg gcaaaggaac atctgccgcc
     2881 gacgccgtgg aagtgcctgc cccagccgcc gtgctgggcg gaccagagcc actgatgcag
     2941 gccaccgcct ggctcaacgc ctactttcac cagcctgagg ccatcgagga gttccctgtg
     3001 ccagccctgc accacccagt gttccagcag gagagcttta cccgccaggt gctgtggaaa
     3061 ctgctgaaag tggtgaagtt cggagaggtc atcagctaca gccacctggc cgccctggcc
     3121 ggcaatcccg ccgccaccgc cgccgtgaaa accgccctga gcggaaatcc cgtgcccatt
     3181 ctgatcccct gccaccgggt ggtgcagggc gacctggacg tggggggcta cgagggcggg
     3241 ctcgccgtga aagagtggct gctggcccac gagggccaca gactgggcaa gcctgggctg
     3301 ggttctagag cagaaaataa ggctaaagtc tcaatggccg gcgctaagcg cgttcccgtg
     3361 accgtaactg ccgcttctaa acccgggttg cgcccggcga ctgctcttgg agatattggc
     3421 aacaaggtgt ctgaagagct gcaggctcgc gtgccactga agcgagaggc taagacgctg
     3481 ggaactggaa agggtacggt taaggccctg ccgaagccag tggagaaagt tcctgtctgc
     3541 gaaccggcta gaatgtataa catgatggag acggagctga agccgccggg cccgcagcaa
     3601 gcttcggggg gcggcggcgg aggaggcaac gccacggcgg cggcgaccgg cggcaaccag
     3661 aagaacagcc cggaccgcgt caagaggccc atgaacgcct tcatggtatg gtcccggggg
     3721 cagcggcgta agatggccca ggagaacccc aagatgcaca actcggagat cagcaagcgc
     3781 ctgggcgcgg agtggaaact tttgtccgag accgagaagc ggccgttcat cgacgaggcc
     3841 aagcggctgc gcgctctgca catgaaggag cacccggatt ataaataccg gccgcggcgg
     3901 aaaaccaaga cgctcatgaa gaaggataag tacacgcttc ccggaggctt gctggccccc
     3961 ggcgggaaca gcatggcgag cggggttggg gtgggcgccg gcctgggtgc gggcgtgaac
     4021 cagcgcatgg acagctacgc gcacatgaac ggctggagca acggcagcta cagcatgatg
     4081 caggagcagc tgggctaccc gcagcacccg ggcctcaacg ctcacggcgc ggcacagatg
     4141 caaccgatgc accgctacga cgtcagcgcc ctgcagtaca actccatgac cagctcgcag
     4201 acctacatga acggctcgcc cacctacagc atgtcctact cgcagcaggg cacccccggt
     4261 atggcgctgg gctccatggg ctctgtggtc aagtccgagg ccagctccag cccccccgtg
     4321 gttacctctt cctcccactc cagggcgccc tgccaggccg gggacctccg ggacatgatc
     4381 agcatgtacc tccccggcgc cgaggtgccg gagcccgctg cgcccagtag actgcacatg
     4441 gcccagcact accagagcgg cccggtgccc ggcacggcca ttaacggcac actgcccctg
     4501 tcgcacatgt gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     4561 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     4621 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     4681 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     4741 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     4801 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     4861 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
     4921 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     4981 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     5041 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     5101 tccccgcctg gaattcgagc tcggtacctt taagaccaat gacttacaag gcagctgtag
     5161 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     5221 gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     5281 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     5341 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     5401 tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt
     5461 tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata
     5521 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5581 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat
     5641 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     5701 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     5761 attccagaag tagtgaggag gcttttttgg aggcctaggg acgtacccaa ttcgccctat
     5821 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     5881 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     5941 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     6001 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     6061 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     6121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     6181 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     6241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     6301 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     6361 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     6421 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
     6481 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     6541 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     6601 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     6661 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     6721 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     6781 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     6841 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     6901 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     6961 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     7021 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     7081 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     7141 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     7201 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     7261 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     7321 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     7381 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     7441 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     7501 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     7561 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     7621 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     7681 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     7741 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     7801 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     7861 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     7921 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     7981 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     8041 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     8101 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     8161 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     8221 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     8281 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     8341 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     8401 gcaaacc
//
