LOCUS       sequence-263603-        8740 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dox-inducible polycistronic lentiviral vector expressing mouse Sox2,
            Klf4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8740)
  AUTHORS   Velychko S, Adachi K, Kim KP, Hou Y, MacCarthy CM, Wu G, Scholer HR
  TITLE     Excluding Oct4 from Yamanaka Cocktail Unleashes the Developmental 
            Potential of iPSCs.
  JOURNAL   Cell Stem Cell. 2019 Oct 30. pii: S1934-5909(19)30423-0. doi: 
            10.1016/j.stem.2019.10.002.
  PUBMED    31708402
REFERENCE   2  (bases 1 to 8740)
  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: "Cell Stem 
            Cell. 2019 Oct 30. pii: S1934-5909(19)30423-0. doi: 
            10.1016/j.stem.2019.10.002."
COMMENT     Sequence Label: pHAGE2-TetOminiCMV-SK
FEATURES             Location/Qualifiers
     source          1..8740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    join(2251..2269,2270..2292,2293..2311,2312..2334,2335..2353,
                     2354..2376,2377..2395,2396..2418,2419..2437,2438..2460,
                     2461..2479,2480..2502,2503..2521)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2554..2592
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2589..2613
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3657..3710
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     misc_feature    5182..5770
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5235..5255)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5653..5664)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5918..6098
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6145..6163)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6231..6335
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6336..6404,6405..7196)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6554..6573)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7367..7955
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8109..8126
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8366..8387
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8402..8407,8408..8425,8426..8432)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8440..8456
                     /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     8445..8467
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8464..8480
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8464..8480
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8530..8549
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtcca
     2221 caccctaact gacacactcg agtttaccac tccctatcag tgatagagaa aagtgaaagt
     2281 cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta
     2341 tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt
     2401 gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac
     2461 tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag
     2521 aaaagtgaaa gtcgagctcg gtacccgggt cgaggtaggc gtgtacggtg ggaggcctat
     2581 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
     2641 gacctccata gaagacaccg ggaccgatcc agcctgcggc cgccatgtat aacatgatgg
     2701 agacggagct gaagccgccg ggcccgcagc aagcttcggg gggcggcggc ggaggaggca
     2761 acgccacggc ggcggcgacc ggcggcaacc agaagaacag cccggaccgc gtcaagaggc
     2821 ccatgaacgc cttcatggta tggtcccggg ggcagcggcg taagatggcc caggagaacc
     2881 ccaagatgca caactcggag atcagcaagc gcctgggcgc ggagtggaaa cttttgtccg
     2941 agaccgagaa gcggccgttc atcgacgagg ccaagcggct gcgcgctctg cacatgaagg
     3001 agcacccgga ttataaatac cggccgcggc ggaaaaccaa gacgctcatg aagaaggata
     3061 agtacacgct tcccggaggc ttgctggccc ccggcgggaa cagcatggcg agcggggttg
     3121 gggtgggcgc cggcctgggt gcgggcgtga accagcgcat ggacagctac gcgcacatga
     3181 acggctggag caacggcagc tacagcatga tgcaggagca gctgggctac ccgcagcacc
     3241 cgggcctcaa cgctcacggc gcggcacaga tgcaaccgat gcaccgctac gacgtcagcg
     3301 ccctgcagta caactccatg accagctcgc agacctacat gaacggctcg cccacctaca
     3361 gcatgtccta ctcgcagcag ggcacccccg gtatggcgct gggctccatg ggctctgtgg
     3421 tcaagtccga ggccagctcc agcccccccg tggttacctc ttcctcccac tccagggcgc
     3481 cctgccaggc cggggacctc cgggacatga tcagcatgta cctccccggc gccgaggtgc
     3541 cggagcccgc tgcgcccagt agactgcaca tggcccagca ctaccagagc ggcccggtgc
     3601 ccggcacggc cattaacggc acactgcccc tgtcgcacat ggcatgcggc tccggcgagg
     3661 gcaggggaag tcttctaaca tgcggggacg tggaggaaaa tcccggccca ctcgagatga
     3721 ggcagccacc tggcgagtct gacatggctg tcagcgacgc tctgctcccg tccttctcca
     3781 cgttcgcgtc cggcccggcg ggaagggaga agacactgcg tccagcaggt gccccgacta
     3841 accgttggcg tgaggaactc tctcacatga agcgacttcc cccacttccc ggccgcccct
     3901 acgacctggc ggcgacggtg gccacagacc tggagagtgg cggagctggt gcagcttgca
     3961 gcagtaacaa cccggccctc ctagcccgga gggagaccga ggagttcaac gacctcctgg
     4021 acctagactt tatcctttcc aactcgctaa cccaccagga atcggtggcc gccaccgtga
     4081 ccacctcggc gtcagcttca tcctcgtctt ccccagcgag cagcggccct gccagcgcgc
     4141 cctccacctg cagcttcagc tatccgatcc gggccggggg tgacccgggc gtggctgcca
     4201 gcaacacagg tggagggctc ctctacagcc gagaatctgc gccacctccc acggccccct
     4261 tcaacctggc ggacatcaat gacgtgagcc cctcgggcgg cttcgtggct gagctcctgc
     4321 ggccggagtt ggacccagta tacattccgc cacagcagcc tcagccgcca ggtggcgggc
     4381 tgatgggcaa gtttgtgctg aaggcgtctc tgaccacccc tggcagcgag tacagcagcc
     4441 cttcggtcat cagtgttagc aaaggaagcc cagacggcag ccaccccgtg gtagtggcgc
     4501 cctacagcgg tggcccgccg cgcatgtgcc ccaagattaa gcaagaggcg gtcccgtcct
     4561 gcacggtcag ccggtcccta gaggcccatt tgagcgctgg accccagctc agcaacggcc
     4621 accggcccaa cacacacgac ttccccctgg ggcggcagct ccccaccagg actaccccta
     4681 cactgagtcc cgaggaactg ctgaacagca gggactgtca ccctggcctg cctcttcccc
     4741 caggattcca tccccatccg gggcccaact accctccttt cctgccagac cagatgcagt
     4801 cacaagtccc ctctctccat tatcaagagc tcatgccacc gggttcctgc ctgccagagg
     4861 agcccaagcc aaagagggga agaaggtcgt ggccccggaa aagaacagcc acccacactt
     4921 gtgactatgc aggctgtggc aaaacctata ccaagagttc tcatctcaag gcacacctgc
     4981 gaactcacac aggcgagaaa ccttaccact gtgactggga cggctgtggg tggaaattcg
     5041 cccgctccga tgaactgacc aggcactacc gcaaacacac agggcaccgg ccctttcagt
     5101 gccagaagtg cgacagggcc ttttccaggt cggaccacct tgccttacac atgaagaggc
     5161 acttttaaat cgatagatcc taatcaacct ctggattaca aaatttgtga aagattgact
     5221 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     5281 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     5341 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     5401 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     5461 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     5521 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     5581 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     5641 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     5701 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     5761 gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     5821 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatca
     5881 cctgcaggac aggcgcgccc tgctttttgc ttgtactggg tctctctggt tagaccagat
     5941 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     6001 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     6061 cctcagaccc ttttagtcag tgtggaaaat ctctagcacc cgggcgatta aggaaagggc
     6121 tagatcattc ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     6181 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     6241 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     6301 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     6361 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     6421 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     6481 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     6541 cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac
     6601 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     6661 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     6721 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     6781 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     6841 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     6901 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     6961 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     7021 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     7081 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     7141 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     7201 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     7261 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     7321 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     7381 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     7441 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     7501 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     7561 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     7621 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     7681 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     7741 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     7801 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     7861 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     7921 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     7981 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     8041 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     8101 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     8161 tccccgcgcg ttggccgatt cattaatgca gcaagctcat ggctgactaa ttttttttat
     8221 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     8281 ttttggaggc ctaggctttt gcaaaaagct ccccgtggca cgacaggttt cccgactgga
     8341 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     8401 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     8461 acacaggaaa cagctatgac atgattacga atttcacaaa taaagcattt ttttcactgc
     8521 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaactgga
     8581 taactcaagc taaccaaaat catcccaaac ttcccacccc ataccctatt accactgcca
     8641 attacctgtg gtttcattta ctctaaacct gtgattcctc tgaattattt tcattttaaa
     8701 gaaattgtat ttgttaaata tgtactacaa acttagtagt
//
