LOCUS       sequence-404825-        6488 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Glis1, Nanog and lin28a by tet-on promoter in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6488)
  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 6488)
  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-GNL
FEATURES             Location/Qualifiers
     source          1..6488
                     /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"
     CDS             2539..2592
                     /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             3517..3573
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     polyA_signal    4339..4420
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4376..4395)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4608..4670
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     rep_origin      complement(4739..5327)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4819..4838)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5498..6289,6290..6358))
                     /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     6121..6140
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6359..6463)
                     /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 actagtgatt ccaccatggt cagcgcccac acacccctgc ccacccactg ccgagcccca
      721 tcgtccatgg gtctgccctc agacctggac tttccagacc gaggcctcac caaccctgca
      781 ccttcctgct accttctggg caatgaaccc atctcagacc tgggtcccca acccgaggcc
      841 cacctccccg agggcagcct gaaacgctgc tgcctcctgg gcctgccccc cacctcttca
      901 gcctcctcct caccctgtgc ctcctcagat atcaatcctg tcatccactc ctcccagaca
      961 gctctagtta gctgtgtaaa tggactccga agcccacctc tgccgggaga cctggggggc
     1021 cctcccaagc ggtcacggcc cgggcctgca tccagtgacg gccaggaggg cagcttgcag
     1081 cttgaagcat gccggaagtc aggcttcctg aagcaggagc ccatggacga gttttcagag
     1141 ctttttgctc cacaccacca gggtttgcca cccccttacc ccttgcctca gttgccaact
     1201 ggccccggcc tcggaggcct agggctgggc ctggcaggta ggatggttgc cggtcggcag
     1261 gcatgccgct gggtggactg ctgcgcagcc tacgagcagc aggaggagct ggtgcggcac
     1321 atcgagaaga gccacatcga ccagcgcaag ggcgaagact tcacctgctt ctgggccggg
     1381 tgtgtgcggc gctacaagcc cttcaatgcc cgctacaagc tgctcatcca catgagggta
     1441 cactcaggcg agaagcccaa caagtgcatg ttcgaaggct gcagtaaagc cttttcccgt
     1501 ctggagaacc tgaagatcca tctgcggagc cacacaggcg agaaaccata cctgtgccag
     1561 cacccaggct gccagaaggc cttcagcaac tccagcgacc gtgccaagca ccaacgcacc
     1621 cacctcgaca cgaagccata tgcttgtcag atccctggct gctccaagcg ctacacggac
     1681 cccagctccc tccgcaagca cgtgaaggcc cactcagcca aagagcagca ggtgcgtaag
     1741 aagctgcaca caggtgccga cccagaggct gatgttctgt ccgagtgtct gtccctgcag
     1801 cagctccaag catccacact gttgccggcc agcagaggga agggcagcca aaccctgagc
     1861 caggagctcc tcccaggtgt gtatcctggc tccgtcaccc cacaaaacgg gcttgcttca
     1921 ggcatcctgt ccccctccca cgatgtccct tccaggcacc acccactgga ggtccccact
     1981 ggttcccacc accacctgtc ccctctgccc acagctgaga gcaccaggga tggcctgggg
     2041 cccagtctcc tttcacccat ggtcagccca ctgaaggggc ttggtccccc accgctacca
     2101 ccagcctccc agagtcagtc tccaggggga cagtcattct ctacagtccc cagcaagcct
     2161 acctacccat ccttccaaag cccaccacct ctgcccagcc cccaaggcta ccaaggcagt
     2221 ttccattcca tccagaactg cttcccctac gctgactgct accgggccac tgagccagca
     2281 gcctccaggg atggactggt gggtgatgcc cacggtttca accccttgcg acccagcaca
     2341 tactccagcc tcagcacacc tttatccgca ccaggctacg agaccctggc agaaacgccg
     2401 tgtcccccag cgctgcagcc acagccagct gaagacctgg tacctagtgg tcctgaggac
     2461 tgtggcttct tccccaatgg ggcctttgac cactgtctga gtcacatccc gtccatctac
     2521 actgacaccg gatccggaga gggcagagga agtctgctaa catgcggtga cgtcgaggag
     2581 aatcctggac ctatgagtgt ggatccagct tgtccccaaa gcttgccttg ctttgaagca
     2641 tccgactgta aagaatcttc acctatgcct gtgatttgtg ggcctgaaga aaactatcca
     2701 tccttgcaaa tgtcttctgc tgagatgcct cacacggaga ctgtctctcc tcttccttcc
     2761 tccatggatc tgcttattca ggacagccct gattcttcca ccagtcccaa aggcaaacaa
     2821 cccacttctg cagagaagag tgtcgcaaaa aaggaagaca aggtcccggt caagaaacag
     2881 aagaccagaa ctgtgttctc ttccacccag ctgtgtgtac tcaatgatag atttcagaga
     2941 cagaaatacc tcagcctcca gcagatgcaa gaactctcca acatcctgaa cctcagctac
     3001 aaacaggtga agacctggtt ccagaaccag agaatgaaat ctaagaggtg gcagaaaaac
     3061 aactggccga agaatagcaa tggtgtgacg cagaaggcct cagcacctac ctaccccagc
     3121 ctttactctt cctaccacca gggatgcctg gtgaacccga ctgggaacct tccaatgtgg
     3181 agcaaccaga cctggaacaa ttcaacctgg agcaaccaga cccagaacat ccagtcctgg
     3241 agcaaccact cctggaacac tcagacctgg tgcacccaat cctggaacaa tcaggcctgg
     3301 aacagtccct tctataactg tggagaggaa tctctgcagt cctgcatgca gttccagcca
     3361 aattctcctg ccagtgactt ggaggctgcc ttggaagctg ctggggaagg ccttaatgta
     3421 atacagcaga ccactaggta ttttagtact ccacaaacca tggatttatt cctaaactac
     3481 tccatgaaca tgcaacctga agacgtggga agcggagcta ctaacttcag cctgctgaag
     3541 caggctggag acgtggagga gaaccctgga cctatgggct ccgtgtccaa ccagcagttt
     3601 gcaggtggct gcgccaaggc ggcagaagag gcgcccgagg aggcgccgga ggacgcggcc
     3661 cgggcggcgg acgagcctca gctgctgcac ggtgcgggca tctgtaagtg gttcaacgtg
     3721 cgcatggggt tcggcttcct gtccatgacc gcccgcgccg gggtcgcgct cgacccccca
     3781 gtggatgtct ttgtgcacca gagtaagctg cacatggaag ggttccggag cttgaaggag
     3841 ggtgaggcag tggagttcac ctttaagaag tcagccaagg gtctggaatc catccgtgtc
     3901 accggacctg gtggagtatt ctgtattggg agtgagaggc ggccaaaagg aaagagcatg
     3961 cagaagcgca gatcaaaagg agacaggtgc tacaactgtg gaggtctaga tcatcatgcc
     4021 aaggaatgca agctgccacc ccagcccaag aagtgccact tctgccagag catcagccat
     4081 atggtagcct catgtccgct gaaggcccag cagggcccta gtgcacaggg aaagccaacc
     4141 tactttcgag aggaagaaga agaaatccac agccctaccc tgctcccgga ggcacagaat
     4201 tgataagata aacgtcgacg atatctctag aggatcataa tcagccatac cacatttgta
     4261 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     4321 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4381 agcatcacaa atttcacaaa taaagcattt ttttcactgc ctcgacacta gttaaaagtt
     4441 ttgttacttt atagaagaaa ttttgagttt ttgttttttt ttaataaata aataaacata
     4501 aataaattgt ttgttgaatt tattattagt atgtaagtgt aaatataata aaacttaata
     4561 tctattcaaa ttaataaata aacctcgata tacagaccga taaaacacat gcgtcaattt
     4621 tacgcatgat tatctttaac gtacgtcaca atatgattat ctttctaggg ttaatcgaca
     4681 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4741 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4801 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4861 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4921 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4981 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5041 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5101 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5161 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5221 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5281 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5341 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5401 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5461 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5521 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5581 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5641 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5701 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5761 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5821 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5881 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5941 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6001 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6061 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6121 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6181 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6241 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6301 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6361 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6421 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6481 tgccacct
//
