LOCUS       sequence-218656-        8403 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of a SOX2 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 8403)
  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 8403)
  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-MD-SOX2
FEATURES             Location/Qualifiers
     source          1..8403
                     /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    4257..4845
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4310..4330)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4728..4739)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4932..5165
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5237..5358
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5274..5293)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5328..5347
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5377..5512
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5544..5563)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5545..5563)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5573..5590)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5573..5589)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5582..5604)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5734..6189
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5821..5840)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6031..6052
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6218..6322
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6323..6391,6392..7183)
                     /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(6541..6560)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7354..7942
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7843..7862
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8096..8113
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8230..8251
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8266..8271,8272..8289,8290..8296)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8304..8320
                     /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     8309..8331
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8328..8344
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8328..8344
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8363..8383
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8365..8383
                     /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 atgtataaca
     3301 tgatggagac ggagctgaag ccgccgggcc cgcagcaagc ttcggggggc ggcggcggag
     3361 gaggcaacgc cacggcggcg gcgaccggcg gcaaccagaa gaacagcccg gaccgcgtca
     3421 agaggcccat gaacgccttc atggtatggt cccgggggca gcggcgtaag atggcccagg
     3481 agaaccccaa gatgcacaac tcggagatca gcaagcgcct gggcgcggag tggaaacttt
     3541 tgtccgagac cgagaagcgg ccgttcatcg acgaggccaa gcggctgcgc gctctgcaca
     3601 tgaaggagca cccggattat aaataccggc cgcggcggaa aaccaagacg ctcatgaaga
     3661 aggataagta cacgcttccc ggaggcttgc tggcccccgg cgggaacagc atggcgagcg
     3721 gggttggggt gggcgccggc ctgggtgcgg gcgtgaacca gcgcatggac agctacgcgc
     3781 acatgaacgg ctggagcaac ggcagctaca gcatgatgca ggagcagctg ggctacccgc
     3841 agcacccggg cctcaacgct cacggcgcgg cacagatgca accgatgcac cgctacgacg
     3901 tcagcgccct gcagtacaac tccatgacca gctcgcagac ctacatgaac ggctcgccca
     3961 cctacagcat gtcctactcg cagcagggca cccccggtat ggcgctgggc tccatgggct
     4021 ctgtggtcaa gtccgaggcc agctccagcc cccccgtggt tacctcttcc tcccactcca
     4081 gggcgccctg ccaggccggg gacctccggg acatgatcag catgtacctc cccggcgccg
     4141 aggtgccgga gcccgctgcg cccagtagac tgcacatggc ccagcactac cagagcggcc
     4201 cggtgcccgg cacggccatt aacggcacac tgcccctgtc gcacatgtga gtcgacaatc
     4261 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     4321 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     4381 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     4441 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     4501 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     4561 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     4621 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct
     4681 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     4741 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     4801 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga attcgagctc
     4861 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     4921 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg
     4981 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     5041 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     5101 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     5161 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     5221 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5281 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5341 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt
     5401 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     5461 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     5521 tgcgtcgaga cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg
     5581 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     5641 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     5701 aacagttgcg cagcctgaat ggcgaatggc gcgacgcgcc ctgtagcggc gcattaagcg
     5761 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     5821 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     5881 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     5941 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     6001 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     6061 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     6121 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     6181 ttacaatttc ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     6241 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     6301 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     6361 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     6421 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     6481 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     6541 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     6601 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     6661 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     6721 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     6781 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     6841 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     6901 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     6961 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     7021 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     7081 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     7141 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     7201 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     7261 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     7321 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     7381 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     7441 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     7501 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     7561 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     7621 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     7681 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     7741 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     7801 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     7861 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     7921 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     7981 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     8041 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     8101 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     8161 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     8221 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     8281 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     8341 tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa agctggagct
     8401 gca
//
