LOCUS       sequence-404877-        7922 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Oct4, Klf4, Sox2 and c-Myc by tet-on promoter in mammalian
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7922)
  AUTHORS   Matsuya S, Fujino K, Imai H, Kusakabe KT, Fujii W, Kano K
  TITLE     Establishment of African pygmy mouse induced pluripotent stem cells 
            using defined doxycycline inducible transcription factors.
  JOURNAL   Sci Rep. 2024 Feb 8;14(1):3204. doi: 10.1038/s41598-024-53687-9.
  PUBMED    38331995
REFERENCE   2  (bases 1 to 7922)
  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: "Sci Rep."; 
            date: "2024-02-8"; volume: "14(1):3204. doi"; pages: " 
            10.1038/s41598-024-53687-9"
COMMENT     Sequence Label: PB-TRE-OKSM
FEATURES             Location/Qualifiers
     source          1..7922
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   13..47
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        336..650
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    344..362
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    380..398
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    415..433
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    451..469
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    487..505
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    522..540
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    558..576
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     623..647
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      661..670
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1756..1809
                     /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"
     CDS             3256..3309
                     /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"
     CDS             4270..4329
                     /codon_start=1
                     /product="2A peptide from equine rhinitis A virus 
                     polyprotein"
                     /label=E2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="QCTNYALLKLAGDVESNPGP"
     CDS             4330..5649
                     /codon_start=1
                     /product="human c-Myc proto-oncogene"
                     /label=c-Myc
                     /translation="MPLNVSFTNRNYDLDYDSVQPYFYCDEEENFYQQQQQSELQPPAP
                     SEDIWKKFELLPTPPLSPSRRSGLCSPSYVAVTPFSLRGDNDGGGGSFSTADQLEMVTE
                     LLGGDMVNQSFICDPDDETFIKNIIIQDCMWSGFSAAAKLVSEKLASYQAARKDSGSPN
                     PARGHSVCSTSSLYLQDLSAAASECIDPSVVFPYPLNDSSSPKSCASQDSSAFSPSSDS
                     LLSSTESSPQGSPEPLVLHEETPPTTSSDSEEEQEDEEEIDVVSVEKRQAPGKRSESGS
                     PSAGGHSKPPHSPLVLKRCHVSTHQHNYAAPPSTRKDYPAAKRVKLDSVRVLRQISNNR
                     KCTSPRSSDTEENVKRRTHNVLERQRRNELKRSFFALRDQIPELENNEKAPKVVILKKA
                     TAYILSVQAEEQKLISEEDLLRKRREQLKHKLEQLRNSCA"
     polyA_signal    5773..5854
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5810..5829)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   6042..6104
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     rep_origin      complement(6173..6761)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6253..6272)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6932..7723,7724..7792))
                     /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     7555..7574
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7793..7897)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgatt aaccctagaa agatagtctg cgtaaaattg acgcatgcat tcttgaaata
       61 ttgctctctc tttctaaata gcgcgaatcc gtcgctgtgc atttaggaca tctcagtcgc
      121 cgcttggagc tcccgtgagg cgtgcttgtc aatgcggtaa gtgtcactga ttttgaacta
      181 taacgaccgc gtgagtcaaa atgacgcatg attatctttt acgtgacttt taagatttaa
      241 ctcatacgat aattatattg ttatttcatg ttctacttac gtgataactt attatatata
      301 tattttcttg ttatagatat caacgggccc atctcgagtt tactccctat cagtgataga
      361 gaacgtatgt cgagtttact ccctatcagt gatagagaac gatgtcgagt ttactcccta
      421 tcagtgatag agaacgtatg tcgagtttac tccctatcag tgatagagaa cgtatgtcga
      481 gtttactccc tatcagtgat agagaacgta tgtcgagttt atccctatca gtgatagaga
      541 acgtatgtcg agtttactcc ctatcagtga tagagaacgt atgtcgaggt aggcgtgtac
      601 ggtgggaggc ctatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagaattc
      661 gccaccatgg cgggacacct ggcttcggat ttcgccttct cgccccctcc aggtggtgga
      721 ggtgatgggc caggggggcc ggagccgggc tgggttgatc ctcggacctg gctaagcttc
      781 caaggccctc ctggagggcc aggaatcggg ccgggggttg ggccaggctc tgaggtgtgg
      841 gggattcccc catgcccccc gccgtatgag ttctgtgggg ggatggcgta ctgtgggccc
      901 caggttggag tggggctagt gccccaaggc ggcttggaga cctctcagcc tgagggcgaa
      961 gcaggagtcg gggtggagag caactccgat ggggcctccc cggagccctg caccgtcacc
     1021 cctggtgccg tgaagctgga gaaggagaag ctggagcaaa acccggagga gtcccaggac
     1081 atcaaagctc tgcagaaaga actcgagcaa tttgccaagc tcctgaagca gaagaggatc
     1141 accctgggat atacacaggc cgatgtgggg ctcaccctgg gggttctatt tgggaaggta
     1201 ttcagccaaa cgaccatctg ccgctttgag gctctgcagc ttagcttcaa gaacatgtgt
     1261 aagctgcggc ccttgctgca gaagtgggtg gaggaagctg acaacaatga aaatcttcag
     1321 gagatatgca aagcagaaac cctcgtgcag gcccgaaaga gaaagcgaac cagtatcgag
     1381 aaccgagtga gaggcaacct ggagaatttg ttcctgcagt gcccgaaacc cacactgcag
     1441 cagatcagcc acatcgccca gcagcttggg ctcgagaagg atgtggtccg agtgtggttc
     1501 tgtaaccggc gccagaaggg caagcgatca agcagcgact atgcacaacg agaggatttt
     1561 gaggctgctg ggtctccttt ctcaggggga ccagtgtcct ttcctctggc cccagggccc
     1621 cattttggta ccccaggcta tgggagccct cacttcactg cactgtactc ctcggtccct
     1681 ttccctgagg gggaagcctt tccccctgtc tccgtcacca ctctgggctc tcccatgcat
     1741 tcaaacggaa gcggagaggg cagaggaagt ctgctaacat gcggtgacgt cgaggagaat
     1801 cctggaccta tgaggcagcc acctggcgag tctgacatgg ctgtcagcga cgcgctgctc
     1861 ccatctttct ccacgttcgc gtctggcccg gcgggaaggg agaagacact gcgtcaagca
     1921 ggtgccccga ataaccgctg gcgggaggag ctctcccaca tgaagcgact tcccccagtg
     1981 cttcccggcc gcccctatga cctggcggcg gcgaccgtgg ccacagacct ggagagcggc
     2041 ggagccggtg cggcttgcgg cggtagcaac ctggcgcccc tacctcggag agagaccgag
     2101 gagttcaacg atctcctgga cctggacttt attctctcca attcgctgac ccatcctccg
     2161 gagtcagtgg ccgccaccgt gtcctcgtca gcgtcagcct cctcttcgtc gtcgccgtcg
     2221 agcagcggcc ctgccagcgc gccctccacc tgcagcttca cctatccgat ccgggccggg
     2281 aacgacccgg gcgtggcgcc gggcggcacg ggcggaggcc tcctctatgg cagggagtcc
     2341 gctccccctc cgacggctcc cttcaacctg gcggacatca acgacgtgag cccctcgggc
     2401 ggcttcgtgg ccgagctcct gcggccagaa ttggacccgg tgtacattcc gccgcagcag
     2461 ccgcagccgc caggtggcgg gctgatgggc aagttcgtgc tgaaggcgtc gctgagcgcc
     2521 cctggcagcg agtacggcag cccgtcggtc atcagcgtca gcaaaggcag ccctgacggc
     2581 agccacccgg tggtggtggc gccctacaac ggcgggccgc cgcgcacgtg ccccaagatc
     2641 aagcaggagg cggtctcttc gtgcacccac ttgggcgctg gaccccctct cagcaatggc
     2701 caccggccgg ctgcacacga cttccccctg gggcggcagc tccccagcag gactaccccg
     2761 accctgggtc ttgaggaagt gctgagcagc agggactgtc accctgccct gccgcttcct
     2821 cccggcttcc atccccaccc ggggcccaat tacccatcct tcctgcccga tcagatgcag
     2881 ccgcaagtcc cgccgctcca ttaccaagag ctcatgccac ccggttcctg catgccagag
     2941 gagcccaagc caaagagggg aagacgatcg tggccccgga aaaggaccgc cacccacact
     3001 tgtgattacg cgggctgcgg caaaacctac acaaagagtt cccatctcaa ggcacacctg
     3061 cgaacccaca caggtgagaa accttaccac tgtgactggg acggctgtgg atggaaattc
     3121 gcccgctcag atgaactgac caggcactac cgtaaacaca cggggcaccg cccgttccag
     3181 tgccaaaaat gcgaccgagc attttccagg tcggaccacc tcgccttaca catgaagagg
     3241 cattttggca gcggcgaggg caggggcagc ctgctgacct gcggcgacgt ggaggagaac
     3301 cccggcccca tgtacaacat gatggagacg gagctgaagc cgccgggccc gcagcaaact
     3361 tcggggggcg gcggcggcaa ctccaccgcg gcggcggccg gcggcaacca gaaaaacagc
     3421 ccggaccgcg tcaagcggcc catgaatgcc ttcatggtgt ggtcccgcgg gcagcggcgc
     3481 aagatggccc aggagaaccc caagatgcac aactcggaga tcagcaagcg cctgggcgcc
     3541 gagtggaaac ttttgtcgga gacggagaag cggccgttca tcgacgaggc taagcggctg
     3601 cgagcgctgc acatgaagga gcacccggat tataaatacc ggccccggcg gaaaaccaag
     3661 acgctcatga agaaggataa gtacacgctg cccggcgggc tgctggcccc cggcggcaat
     3721 agcatggcga gcggggtcgg ggtgggcgcc ggcctgggcg cgggcgtgaa ccagcgcatg
     3781 gacagttacg cgcacatgaa cggctggagc aacggcagct acagcatgat gcaggaccag
     3841 ctgggctacc cgcagcaccc gggcctcaat gcgcacggcg cagcgcagat gcagcccatg
     3901 caccgctacg acgtgagcgc cctgcagtac aactccatga ccagctcgca gacctacatg
     3961 aacggctcgc ccacctacag catgtcctac tcgcagcagg gcacccctgg catggctctt
     4021 ggctccatgg gttcggtggt caagtccgag gccagctcca gcccccctgt ggttacctct
     4081 tcctcccact ccagggcgcc ctgccaggcc ggggacctcc gggacatgat cagcatgtat
     4141 ctccccggcg ccgaggtgcc ggaacccgcc gcccccagca gacttcacat gtcccagcac
     4201 taccagagcg gcccggtgcc cggcacggcc attaacggca cactgcccct ctcacacatg
     4261 ggtagtgggc aatgtactaa ctacgctttg ttgaaactcg ctggcgatgt tgaaagtaac
     4321 cccggtccta tgcccctcaa cgttagcttc accaacagga actatgacct cgactacgac
     4381 tcggtgcagc cgtatttcta ctgcgacgag gaggagaact tctaccagca gcagcagcag
     4441 agcgagctgc agcccccggc gcccagcgag gatatctgga agaaattcga gctgctgccc
     4501 accccgcccc tgtcccctag ccgccgctcc gggctctgct cgccctccta cgttgcggtc
     4561 acacccttct cccttcgggg agacaacgac ggcggtggcg ggagcttctc cacggccgac
     4621 cagctggaga tggtgaccga gctgctggga ggagacatgg tgaaccagag tttcatctgc
     4681 gacccggacg acgagacctt catcaaaaac atcatcatcc aggactgtat gtggagcggc
     4741 ttctcggccg ccgccaagct cgtctcagag aagctggcct cctaccaggc tgcgcgcaaa
     4801 gacagcggca gcccgaaccc cgcccgcggc cacagcgtct gctccacctc cagcttgtac
     4861 ctgcaggatc tgagcgccgc cgcctcagag tgcatcgacc cctcggtggt cttcccctac
     4921 cctctcaacg acagcagctc gcccaagtcc tgcgcctcgc aagactccag cgccttctct
     4981 ccgtcctcgg attctctgct ctcctcgacg gagtcctccc cgcagggcag ccccgagccc
     5041 ctggtgctcc atgaggagac accgcccacc accagcagcg actctgagga ggaacaagaa
     5101 gatgaggaag aaatcgatgt tgtttctgtg gaaaagaggc aggctcctgg caaaaggtca
     5161 gagtctggat caccttctgc tggaggccac agcaaacctc ctcacagccc actggtcctc
     5221 aagaggtgcc acgtctccac acatcagcac aactacgcag cgcctccctc cactcggaag
     5281 gactatcctg ctgccaagag ggtcaagttg gacagtgtca gagtcctgag acagatcagc
     5341 aacaaccgaa aatgcaccag ccccaggtcc tcggacaccg aggagaatgt caagaggcga
     5401 acacacaacg tcttggagcg ccagaggagg aacgagctaa aacggagctt ttttgccctg
     5461 cgtgaccaga tcccggagtt ggaaaacaat gaaaaggccc ccaaggtagt tatccttaaa
     5521 aaagccacag catacatcct gtccgtccaa gcagaggagc aaaagctcat ttctgaagag
     5581 gacttgttgc ggaaacgacg agaacagttg aaacacaaac ttgaacagct acggaactct
     5641 tgtgcgtaat agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt
     5701 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     5761 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5821 acaaatttca caaataaagc atttttttca ctgcctcgac actagttaaa agttttgtta
     5881 ctttatagaa gaaattttga gtttttgttt ttttttaata aataaataaa cataaataaa
     5941 ttgtttgttg aatttattat tagtatgtaa gtgtaaatat aataaaactt aatatctatt
     6001 caaattaata aataaacctc gatatacaga ccgataaaac acatgcgtca attttacgca
     6061 tgattatctt taacgtacgt cacaatatga ttatctttct agggttaatc gacatgtgag
     6121 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6181 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6241 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6301 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6361 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6421 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6481 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6541 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6601 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6661 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6721 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6781 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6841 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6901 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6961 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     7021 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     7081 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     7141 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7201 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7261 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7321 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7381 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7441 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7501 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7561 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7621 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7681 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7741 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7801 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7861 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     7921 ct
//
