LOCUS       sequence-98026-e        7397 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral DOX-inducible expression vector for human NANOG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7397)
  AUTHORS   Theunissen TW, Powell BE, Wang H, Mitalipova M, Faddah DA, Reddy J, 
            Fan ZP, Maetzel D, Ganz K, Shi L, Lungjangwa T, Imsoonthornruksa S, 
            Stelzer Y, Rangarajan S, D'Alessio A, Zhang J, Gao Q, Dawlaty MM, 
            Young RA, Gray NS, Jaenisch R
  TITLE     Systematic identification of culture conditions for induction and 
            maintenance of naive human pluripotency.
  JOURNAL   Cell Stem Cell. 2014 Oct 2;15(4):471-87. doi: 
            10.1016/j.stem.2014.07.002. Epub 2014 Jul 24.
  PUBMED    25090446
REFERENCE   2  (bases 1 to 7397)
  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: 
            "10.1016/j.stem.2014.07.002"; journalName: "Cell Stem Cell"; date: 
            "2014-10-2- 2"; volume: "15"; issue: "4"; pages: "471-87"
COMMENT     Sequence Label: FUW-tetO-loxP-hNANOG
FEATURES             Location/Qualifiers
     source          1..7397
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        6..385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..585
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     540..560
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             603..783
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    830..955
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1448..1681
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1866..1910
                     /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    2208..2325
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    join(2384..2402,2403..2425,2426..2444,2445..2467,2468..2486,
                     2487..2509,2510..2528,2529..2551,2552..2570,2571..2593,
                     2594..2612,2613..2635,2636..2654)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2687..2724
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2721..2745
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     misc_feature    3786..4374
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3839..3859)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4257..4268)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4377..4393)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4378..4394)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    4898..4931
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             4945..5125
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5151..5168)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5157..5268
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(5414..6002)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5494..5513)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6173..6964,6965..7033))
                     /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     6796..6815
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7034..7138)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7213..7232)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
       61 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      121 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      181 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      361 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      421 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      481 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      541 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagcgcgt tttgcctgta
      601 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
      661 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
      721 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
      781 gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg
      841 caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac
      901 gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat
      961 taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa
     1021 aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa
     1081 tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc
     1141 ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg
     1201 tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga
     1261 gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag
     1321 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac
     1381 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag
     1441 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag
     1501 cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga
     1561 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca
     1621 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc
     1681 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta
     1741 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca
     1801 gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc
     1861 aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt
     1921 ttaacataac aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg
     1981 taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt
     2041 caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa
     2101 tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg aacggatcgg
     2161 cactgcgtgc gccaattctg cagacaaatg gcagtattca tccacaattt taaaagaaaa
     2221 ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata
     2281 caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg
     2341 gacagcagag atccagtttg gttaatgatc tcgagtttac cactccctat cagtgataga
     2401 gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt
     2461 ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt
     2521 gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag
     2581 tcgagtttac cactccctat cagtgataga gaaaagtgaa agtcgagttt accactccct
     2641 atcagtgata gagaaaagtg aaagtcgagc tcggtacccg ggtcgagtag gcgtgtacgg
     2701 tgggaggcct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
     2761 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg cggccccgaa
     2821 ttcactagtg attcatgagt gtggatccag cttgtcccca aagcttgcct tgctttgaag
     2881 catccgactg taaagaatct tcacctatgc ctgtgatttg tgggcctgaa gaaaactatc
     2941 catccttgca aatgtcttct gctgagatgc ctcacacgga gactgtctct cctcttcctt
     3001 cctccatgga tctgcttatt caggacagcc ctgattcttc caccagtccc aaaggcaaac
     3061 aacccacttc tgcagagaag agtgtcgcaa aaaaggaaga caaggtcccg gtcaagaaac
     3121 agaagaccag aactgtgttc tcttccaccc agctgtgtgt actcaatgat agatttcaga
     3181 gacagaaata cctcagcctc cagcagatgc aagaactctc caacatcctg aacctcagct
     3241 acaaacaggt gaagacctgg ttccagaacc agagaatgaa atctaagagg tggcagaaaa
     3301 acaactggcc gaagaatagc aatggtgtga cgcagaaggc ctcagcacct acctacccca
     3361 gcctttactc ttcctaccac cagggatgcc tggtgaaccc gactgggaac cttccaatgt
     3421 ggagcaacca gacctggaac aattcaacct ggagcaacca gacccagaac atccagtcct
     3481 ggagcaacca ctcctggaac actcagacct ggtgcaccca atcctggaac aatcaggcct
     3541 ggaacagtcc cttctataac tgtggagagg aatctctgca gtcctgcatg cagttccagc
     3601 caaattctcc tgccagtgac ttggaggctg ccttggaagc tgctggggaa ggccttaatg
     3661 taatacagca gaccactagg tattttagta ctccacaaac catggattta ttcctaaact
     3721 actccatgaa catgcaacct gaagacgtgt gaagatgaat cgaattcgat atcaagctta
     3781 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     3841 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     3901 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     3961 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     4021 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     4081 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     4141 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     4201 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     4261 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     4321 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     4381 accgtcgacc tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta
     4441 ccaatgctga ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca
     4501 cacctcaggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     4561 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg
     4621 atctgtggat ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc
     4681 cagggatcag atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag
     4741 agaaggtaga agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc
     4801 atgggatgga tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat
     4861 ttcatcacat ggcccgagag ctgcatccgg actcgagata acttcgtata atgtatgcta
     4921 tacgaagtta ttccggactg tactgggtct ctctggttag accagatctg agcctgggag
     4981 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     5041 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     5101 tagtcagtgt ggaaaatctc tagcagggcc cgtttaaacc cgctgatcag cctcgactgt
     5161 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     5221 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     5281 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     5341 agacaatagc aggcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5401 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5461 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5521 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5581 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5641 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5701 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5761 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5821 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5881 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5941 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6001 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6061 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6121 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6181 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6241 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6301 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6361 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6421 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6481 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6541 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6601 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6661 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6721 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6781 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6841 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6901 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6961 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7021 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7081 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7141 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     7201 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     7261 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     7321 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
     7381 tgtacgggcc agatata
//
