LOCUS       sequence-220070-        8049 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express reverse tetracycline inducible rtTA transcriptional 
            activator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8049)
  AUTHORS   Ye L, Chang JC, Lin C, Qi Z, Yu J, Kan YW
  TITLE     Generation of induced pluripotent stem cells using site-specific 
            integration with phage integrase.
  JOURNAL   Proc Natl Acad Sci U S A. 2010 Nov 9;107(45):19467-72. Epub 2010 Oct
            25.
  PUBMED    20974949
REFERENCE   2  (bases 1 to 8049)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2010-11-9"; volume: "107(45)"; pages: 
            "19467-72. Epub 2010 Oct 25"
COMMENT     Sequence Label: pCAG-Tet-on Advanced -BGH-PA
FEATURES             Location/Qualifiers
     source          1..8049
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        3..382
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..660
                     /label=chicken beta-actin promoter
     intron          661..1669
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1593..1615
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1677..1696
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(1727..2344,2345..2350,2351..2386,2387..2389,2390..2425,
                     2426..2428,2429..2464,2465..2473)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             2510..2551
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2569..2586)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2575..2799
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3010..3029)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(3123..3140)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3294..3882)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3374..3393)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4053..4844,4845..4913))
                     /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     4676..4695
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4914..5018)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5086..5104
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     polyA_signal    5131..5265
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5155..5174)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5209..5228
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5690..5710)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          5840..5905
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(6325..7119)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6436..6455)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7046..7065
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(7480..7809)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7494..7629
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7794..7814
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(7852..7874)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8015..8034
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tcgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
       61 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      121 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      181 gactttccat tgacgtcaat gggtggacta tttacggtaa actgcccact tggcagtaca
      241 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      301 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      361 attagtcatc gctattacca tgggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg
      541 ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
      601 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
      661 ggagtcgctg cgttgccttc gccccgtgcc ccgctccgcg ccgcctcgcg ccgcccgccc
      721 cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg
      781 ggctgtaatt agcgcttggt ttaatgacgg ctcgtttctt ttctgtggct gcgtgaaagc
      841 cttaaagggc tccgggaggg ccctttgtgc gggggggagc ggctcggggg gtgcgtgcgt
      901 gtgtgtgtgc gtggggagcg ccgcgtgcgg cccgcgctgc ccggcggctg tgagcgctgc
      961 gggcgcggcg cggggctttg tgcgctccgc gtgtgcgcga ggggagcgcg gccgggggcg
     1021 gtgccccgcg gtgcgggggg gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt
     1081 gggggggtga gcagggggtg tgggcgcggc ggtcgggctg taaccccccc ctgcaccccc
     1141 ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtgcg gggcgtggcg
     1201 cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc
     1261 cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg ccccggagcg ccggcggctg
     1321 tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg
     1381 acttcctttg tcccaaatct ggcggagccg aaatctggga ggcgccgccg caccccctct
     1441 agcgggcgcg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg gagggccttc
     1501 gtgcgtcgcc gcgccgccgt ccccttctcc atctccagcc tcggggctgc cgcaggggga
     1561 cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg
     1621 ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc tcctgggcaa
     1681 cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcaccatgt ctagactgga
     1741 caagagcaaa gtcataaacg gcgctctgga attactcaat ggagtcggta tcgaaggcct
     1801 gacgacaagg aaactcgctc aaaagctggg agttgagcag cctaccctgt actggcacgt
     1861 gaagaacaag cgggccctgc tcgatgccct gccaatcgag atgctggaca ggcatcatac
     1921 ccacttctgc cccctggaag gcgagtcatg gcaagacttt ctgcggaaca acgccaagtc
     1981 attccgctgt gctctcctct cacatcgcga cggggctaaa gtgcatctcg gcacccgccc
     2041 aacagagaaa cagtacgaaa ccctggaaaa tcagctcgcg ttcctgtgtc agcaaggctt
     2101 ctccctggag aacgcactgt acgctctgtc cgccgtgggc cactttacac tgggctgcgt
     2161 attggaggaa caggagcatc aagtagcaaa agaggaaaga gagacaccta ccaccgattc
     2221 tatgccccca cttctgagac aagcaattga gctgttcgac cggcagggag ccgaacctgc
     2281 cttccttttc ggcctggaac taatcatatg tggcctggag aaacagctaa agtgcgaaag
     2341 cggcgggccg gccgacgccc ttgacgattt tgacttagac atgctcccag ccgatgccct
     2401 tgacgacttt gaccttgata tgctgcctgc tgacgctctt gacgattttg accttgacat
     2461 gctccccggg taactaagta aggatctggc cggctgggcc cgtttcgaag gtaagcctat
     2521 ccctaaccct ctcctcggtc tcgattctac gcgaaacccg ctgatcagcc tcgactgtgc
     2581 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     2641 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     2701 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     2761 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     2821 gtggcggtaa tacggttatc cacagaatca gaagggcgaa ttccagcaca ctggcggccg
     2881 ttactagtgg atccgagctc ggtaccaagc ttggcgagat tttcaggagc taaggaagct
     2941 aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa
     3001 gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg
     3061 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3121 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3181 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3241 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3301 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3361 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3421 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3481 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3541 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3601 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3661 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3721 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3781 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3841 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3901 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3961 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4021 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4081 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4141 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     4201 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     4261 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     4321 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     4381 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4441 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4501 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4561 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4621 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat
     4681 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4741 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4801 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4861 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4921 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4981 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5041 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     5101 aggccctttc gtcttcactc gaggtcgagg gatccagaca tgataagata cattgatgag
     5161 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     5221 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     5281 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     5341 ctctacaaat gtggtatggc tgattatgat ctctagtcaa ggcactatac atcaaatatt
     5401 ccttattaac ccctttacaa attaaaaagc taaaggtaca caatttttga gcatagttat
     5461 taatagcaga cactctatgc ctgtgtggag taagaaaaaa cagtatgtta tgattataac
     5521 tgttatgcct acttataaag gttacagaat atttttccat aattttcttg tatagcagtg
     5581 cagctttttc ctttgtggtg taaatagcaa agcaagcaag agttctatta ctaaacacag
     5641 catgactcaa aaaacttagc aattctgaag gaaagtcctt ggggtcttct acctttctct
     5701 tcttttttgg aggagtagaa tgttgagagt cagcagtagc ctcatcatca ctagatggca
     5761 tttcttctga gcaaaacagg ttttcctcat taaaggcatt ccaccactgc tcccattcat
     5821 cagttccata ggttggaatc taaaatacac aaacaattag aatcagtagt ttaacacatt
     5881 atacacttaa aaattttata tttaccttag agctttaaat ctctgtaggt agtttgtcca
     5941 attatgtcac accacagaag taaggttcct tcacaaagat ccggaccaaa gcggccatcg
     6001 tgcctcccca ctcctgcagt tcgggggcat ggatgcgcgg atagccgctg ctggtttcct
     6061 ggatgccgac ggatttgcac tgccggtaga actccgcgag gtcgtccagc ctcaggcagc
     6121 agctgaacca actcgcgagg ggatcgagcc cggggtgggc gaagaactcc agcatgagat
     6181 ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt
     6241 catagaaggc ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg
     6301 tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat
     6361 gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc
     6421 gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac
     6481 acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg
     6541 caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc gcgccttgag
     6601 cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc
     6661 gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc
     6721 gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga
     6781 tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa
     6841 tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc
     6901 cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga
     6961 caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc
     7021 atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc
     7081 ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt
     7141 ctcttgatca gatcttgatc ccctgcgcca tcagatcctt ggcggcaaga aagccatcca
     7201 gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca attccggttc
     7261 gcttgctgtc cataaaaccg cccagtctag ctatcgccat gtaagcccac tgcaagctac
     7321 ctgctttctc tttgcgcttg cgttttccct tgtccagata gcccagtagc tgacattcat
     7381 ccggggtcag caccgtttct gcggactggc tttctacgtg ttccgcttcc tttagcagcc
     7441 cttgcgccct gagtgcttgc ggcagcgtga agctagcttt ttgcaaaagc ctaggcctcc
     7501 aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc
     7561 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
     7621 gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct
     7681 ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt
     7741 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa
     7801 ctgacacaca ttccacagct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     7861 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     7921 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg
     7981 tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga
     8041 gtgcacctc
//
