LOCUS       sequence-379818-        9456 bp    DNA     circular SYN 15-APR-2025
DEFINITION  tetO-KS* (Dox-inducible reprogramming lentiviral vector expressing 
            mouse Klf4 + super-Sox).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9456)
  AUTHORS   MacCarthy CM, Wu G, Malik V, Menuchin-Lasowski Y, Velychko T, Keshet
            G, Fan R, Bedzhov I, Church GM, Jauch R, Cojocaru V, Scholer HR, 
            Velychko S
  TITLE     Highly cooperative chimeric super-SOX induces naive pluripotency 
            across species.
  JOURNAL   Cell Stem Cell. 2024 Jan 4;31(1):127-147.e9. doi: 
            10.1016/j.stem.2023.11.010. Epub 2023 Dec 22.
  PUBMED    38141611
REFERENCE   2  (bases 1 to 9456)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.stem.2023.11.010"; journalName: "Cell Stem Cell"; date: 
            "2024-01-4- 4"; volume: "31"; issue: "1"; pages: "127-147.e9"
COMMENT     Sequence Label: pHAGE2-TetOminiCMV-Klf4-IRES-Sox2-17
FEATURES             Location/Qualifiers
     source          1..9456
                     /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"
     primer_bind     2589..2613
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2679..2688
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    4113..4680
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4130..4682
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4274..4291)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4501..4520
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     misc_feature    5898..6486
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5951..5971)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6369..6380)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6634..6814
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6861..6879)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6947..7051
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7052..7120,7121..7912)
                     /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(7270..7289)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8083..8671
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8825..8842
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9082..9103
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9118..9123,9124..9141,9142..9148)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9156..9172
                     /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     9161..9183
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9180..9196
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9180..9196
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9246..9265
                     /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 atgtacggtg ggaggcctat
     2581 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
     2641 gacctccata gaagacaccg ggaccgatcc agcctgcggc cgccatggct gtcagcgacg
     2701 ctctgctccc gtccttctcc acgttcgcgt ccggcccggc gggaagggag aagacactgc
     2761 gtccagcagg tgccccgact aaccgttggc gtgaggaact ctctcacatg aagcgacttc
     2821 ccccacttcc cggccgcccc tacgacctgg cggcgacggt ggccacagac ctggagagtg
     2881 gcggagctgg tgcagcttgc agcagtaaca acccggccct cctagcccgg agggagaccg
     2941 aggagttcaa cgacctcctg gacctagact ttatcctttc caactcgcta acccaccagg
     3001 aatcggtggc cgccaccgtg accacctcgg cgtcagcttc atcctcgtct tccccagcga
     3061 gcagcggccc tgccagcgcg ccctccacct gcagcttcag ctatccgatc cgggccgggg
     3121 gtgacccggg cgtggctgcc agcaacacag gtggagggct cctctacagc cgagaatctg
     3181 cgccacctcc cacggccccc ttcaacctgg cggacatcaa tgacgtgagc ccctcgggcg
     3241 gcttcgtggc tgagctcctg cggccggagt tggacccagt atacattccg ccacagcagc
     3301 ctcagccgcc aggtggcggg ctgatgggca agtttgtgct gaaggcgtct ctgaccaccc
     3361 ctggcagcga gtacagcagc ccttcggtca tcagtgttag caaaggaagc ccagacggca
     3421 gccaccccgt ggtagtggcg ccctacagcg gtggcccgcc gcgcatgtgc cccaagatta
     3481 agcaagaggc ggtcccgtcc tgcacggtca gccggtccct agaggcccat ttgagcgctg
     3541 gaccccagct cagcaacggc caccggccca acacacacga cttccccctg gggcggcagc
     3601 tccccaccag gactacccct acactgagtc ccgaggaact gctgaacagc agggactgtc
     3661 accctggcct gcctcttccc ccaggattcc atccccatcc ggggcccaac taccctcctt
     3721 tcctgccaga ccagatgcag tcacaagtcc cctctctcca ttatcaagag ctcatgccac
     3781 cgggttcctg cctgccagag gagcccaagc caaagagggg aagaaggtcg tggccccgga
     3841 aaagaacagc cacccacact tgtgactatg caggctgtgg caaaacctat accaagagtt
     3901 ctcatctcaa ggcacacctg cgaactcaca caggcgagaa accttaccac tgtgactggg
     3961 acggctgtgg gtggaaattc gcccgctcgg atgaactgac caggcactac cgcaaacaca
     4021 cagggcaccg gccctttcag tgccagaagt gcgacagggc cttttccagg tcggaccacc
     4081 ttgccttaca catgaagagg cacttttaaa gatccctccc ccccccctaa cgttactggc
     4141 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg
     4201 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct
     4261 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca
     4321 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg
     4381 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct
     4441 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
     4501 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
     4561 atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa
     4621 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa
     4681 tatggccaca catatgatgt ataacatgat ggagacggag ctgaagccgc cgggcccgca
     4741 gcaagcttcg gggggcggcg gcggaggagg caacgccacg gcggcggcaa ccggcggcaa
     4801 ccagaagaac agcccggacc gcgtcaagag gcccatgaac gccttcatgg tatggtcccg
     4861 ggggcagcgg cgtaagatgg cccaggagaa ccccaagatg cacaactcgg agatcagcaa
     4921 gcgcctgggc gcggagtgga aagcgttgac cttggcagag aagcggccgt tcatcgacga
     4981 ggccaagcgg ctgcgcgtgc tgcatatgca ggaccacccc aactacaagt accggccgcg
     5041 gcggcgcaag caggtgaagc gcatgaagcg ggtggaggga ggcttcctgc acgctctcgt
     5101 cgagccccag gccggcgcgc ttggtcccga gggcggccgc gtggccatgg atggcctggg
     5161 tctgcctttc ccggagccgg gctatccggc cggtcctccg ctgatgtctc cgcacatggg
     5221 cccccactat cgggactgcc agggactggg cgctcccgcg ctcgacggct accctctgcc
     5281 cactccggac acatccccgc tggatggcgt ggagcaggac ccggctttct ttgcagcccc
     5341 gctgccaggg gactgcccgg cggccggcac ctacacttac gctccagtct cggactatgc
     5401 agtgtccgta gagccgcccg ctggccccat gcgagtgggg ccggacccct cgggccctgc
     5461 gatgccgggg atcctggcgc cccccagcgc tctgcacctg tactacggcg cgatgggctc
     5521 gcccgccgca agtgcggggc gcggtttcca cgcgcaaccc cagcagccgc tgcaaccgca
     5581 ggcaccgccc ccgccaccgc agcagcagca cccagcgcac ggccccgggc aaccttcgcc
     5641 ccctcccgag gctctgccct gccgggatgg cacggaatcc aaccagccca ctgagctcct
     5701 aggggaggtg gaccgcacgg aatttgaaca gtatctgccc tttgtgtata agcccgagat
     5761 gggtcttccc taccagggac acgactgcgg agtgaacctc tcagacagcc acggagccat
     5821 ttcctccgtg gtgtccgacg ctagctcagc ggtctactat tgcaactacc ccgacatttg
     5881 ataaatcgat agatcctaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     5941 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     6001 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     6061 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     6121 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     6181 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     6241 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     6301 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     6361 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     6421 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     6481 ccccgcctgg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     6541 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca agatcacctg
     6601 caggacaggc gcgccctgct ttttgcttgt actgggtctc tctggttaga ccagatctga
     6661 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     6721 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     6781 agaccctttt agtcagtgtg gaaaatctct agcacccggg cgattaagga aagggctaga
     6841 tcattcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     6901 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     6961 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     7021 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     7081 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     7141 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     7201 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     7261 taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg
     7321 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     7381 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     7441 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     7501 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     7561 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     7621 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     7681 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     7741 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     7801 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     7861 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     7921 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     7981 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     8041 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     8101 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     8161 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     8221 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     8281 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     8341 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     8401 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     8461 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     8521 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     8581 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     8641 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     8701 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     8761 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     8821 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     8881 cgcgcgttgg ccgattcatt aatgcagcaa gctcatggct gactaatttt ttttatttat
     8941 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     9001 ggaggcctag gcttttgcaa aaagctcccc gtggcacgac aggtttcccg actggaaagc
     9061 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     9121 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     9181 aggaaacagc tatgacatga ttacgaattt cacaaataaa gcattttttt cactgcattc
     9241 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatca actggataac
     9301 tcaagctaac caaaatcatc ccaaacttcc caccccatac cctattacca ctgccaatta
     9361 cctgtggttt catttactct aaacctgtga ttcctctgaa ttattttcat tttaaagaaa
     9421 ttgtatttgt taaatatgta ctacaaactt agtagt
//
