LOCUS       sequence-263722-        9228 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dox-inducible polycistronic reprogramming lentiviral vector 
            expressing mouse Klf4, Sox2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9228)
  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 9228)
  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-KS
FEATURES             Location/Qualifiers
     source          1..9228
                     /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"
     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    5670..6258
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5723..5743)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6141..6152)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6406..6586
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6633..6651)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6719..6823
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6824..6892,6893..7684)
                     /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(7042..7061)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7855..8443
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8597..8614
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8854..8875
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8890..8895,8896..8913,8914..8920)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8928..8944
                     /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     8933..8955
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8952..8968
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8952..8968
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9018..9037
                     /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 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 gcccgctccg atgaactgac caggcactac cgcaaacaca
     4021 cagggcaccg gccctttcag tgccagaagt gtgacagggc 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 gcggcggcga ccggcggcaa
     4801 ccagaagaac agcccggacc gcgtcaagag gcccatgaac gccttcatgg tatggtcccg
     4861 ggggcagcgg cgtaagatgg cccaggagaa ccccaagatg cacaactcgg agatcagcaa
     4921 gcgcctgggc gcggagtgga aacttttgtc cgagaccgag aagcggccgt tcatcgacga
     4981 ggccaagcgg ctgcgcgctc tgcacatgaa ggagcacccg gattataaat accggccgcg
     5041 gcggaaaacc aagacgctca tgaagaagga taagtacacg cttcccggag gcttgctggc
     5101 ccccggcggg aacagcatgg cgagcggggt tggggtgggc gccggcctgg gtgcgggcgt
     5161 gaaccagcgc atggacagct acgcgcacat gaacggctgg agcaacggca gctacagcat
     5221 gatgcaggag cagctgggct acccgcagca cccgggcctc aacgctcacg gcgcggcaca
     5281 gatgcaaccg atgcaccgct acgacgtcag cgccctgcag tacaactcca tgaccagctc
     5341 gcagacctac atgaacggct cgcccaccta cagcatgtcc tactcgcagc agggcacccc
     5401 cggtatggcg ctgggctcca tgggctctgt ggtcaagtcc gaggccagct ccagcccccc
     5461 cgtggttacc tcttcctccc actccagggc gccctgccag gccggggacc tccgggacat
     5521 gatcagcatg tacctccccg gcgccgaggt gccggagccc gctgcgccca gtagactgca
     5581 catggcccag cactaccaga gcggcccggt gcccggcacg gccattaacg gcacactgcc
     5641 cctgtcgcac atgtaaatcg atagatccta atcaacctct ggattacaaa atttgtgaaa
     5701 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     5761 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     5821 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     5881 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     5941 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     6001 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     6061 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     6121 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     6181 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     6241 tttgggccgc ctccccgcct ggtaccttta agaccaatga cttacaaggc agctgtagat
     6301 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
     6361 caagatcacc tgcaggacag gcgcgccctg ctttttgctt gtactgggtc tctctggtta
     6421 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     6481 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     6541 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcacccg ggcgattaag
     6601 gaaagggcta gatcattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     6661 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     6721 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     6781 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     6841 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     6901 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     6961 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     7021 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     7081 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     7141 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     7201 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     7261 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     7321 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     7381 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     7441 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     7501 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     7561 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     7621 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     7681 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     7741 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     7801 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     7861 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     7921 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     7981 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     8041 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     8101 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     8161 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     8221 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     8281 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     8341 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     8401 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     8461 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     8521 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     8581 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     8641 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc aagctcatgg ctgactaatt
     8701 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     8761 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc ccgtggcacg acaggtttcc
     8821 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     8881 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     8941 acaatttcac acaggaaaca gctatgacat gattacgaat ttcacaaata aagcattttt
     9001 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     9061 caactggata actcaagcta accaaaatca tcccaaactt cccaccccat accctattac
     9121 cactgccaat tacctgtggt ttcatttact ctaaacctgt gattcctctg aattattttc
     9181 attttaaaga aattgtattt gttaaatatg tactacaaac ttagtagt
//
