LOCUS       sequence-220023-        8056 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 8056)
  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 8056)
  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..8056
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        3..382
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          660..1676
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1600..1622
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1684..1703
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(1734..2351,2352..2357,2358..2393,2394..2396,2397..2432,
                     2433..2435,2436..2471,2472..2480)
                     /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             2517..2558
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2576..2593)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2582..2806
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3017..3036)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(3130..3147)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3301..3889)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3381..3400)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4060..4851,4852..4920))
                     /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     4683..4702
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4921..5025)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5093..5111
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     polyA_signal    5138..5272
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5162..5181)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5216..5235
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5697..5717)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          5847..5912
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(6332..7126)
                     /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(6443..6462)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7053..7072
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(7487..7816)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7501..7636
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7801..7821
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(7859..7881)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8022..8041
                     /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 gggtggagta tttacggtaa actgcccact tggcagtaca
      241 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      301 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      361 attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat
      421 ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc
      481 gatgggggcg gggggggggg gggsgcgcgc crggsggggs gggggggggs gggggggggg
      541 gsggggsgrg gsggrgrggk gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
      601 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
      661 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
      721 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      781 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
      841 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct
      961 gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg
     1021 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
     1081 gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc
     1141 cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg
     1201 gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg
     1261 ggccgcctcg ggccggggrg ggctcggggg rggggcgcgg cggcccccgg agcgccggcg
     1321 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
     1381 agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc
     1441 cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg
     1501 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc
     1561 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga
     1621 ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc
     1681 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattc accatgtcta
     1741 gactggacaa gagcaaagtc ataaacggcg ctctggaatt actcaatgga gtcggtatcg
     1801 aaggcctgac gacaaggaaa ctcgctcaaa agctgggagt tgagcagcct accctgtact
     1861 ggcacgtgaa gaacaagcgg gccctgctcg atgccctgcc aatcgagatg ctggacaggc
     1921 atcataccca cttctgcccc ctggaaggcg agtcatggca agactttctg cggaacaacg
     1981 ccaagtcatt ccgctgtgct ctcctctcac atcgcgacgg ggctaaagtg catctcggca
     2041 cccgcccaac agagaaacag tacgaaaccc tggaaaatca gctcgcgttc ctgtgtcagc
     2101 aaggcttctc cctggagaac gcactgtacg ctctgtccgc cgtgggccac tttacactgg
     2161 gctgcgtatt ggaggaacag gagcatcaag tagcaaaaga ggaaagagag acacctacca
     2221 ccgattctat gcccccactt ctgagacaag caattgagct gttcgaccgg cagggagccg
     2281 aacctgcctt ccttttcggc ctggaactaa tcatatgtgg cctggagaaa cagctaaagt
     2341 gcgaaagcgg cgggccggcc gacgcccttg acgattttga cttagacatg ctcccagccg
     2401 atgcccttga cgactttgac cttgatatgc tgcctgctga cgctcttgac gattttgacc
     2461 ttgacatgct ccccgggtaa ctaagtaagg atctggccgg ctgggcccgt ttcgaaggta
     2521 agcctatccc taaccctctc ctcggtctcg attctacgcg aaacccgctg atcagcctcg
     2581 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     2641 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     2701 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     2761 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     2821 agaaccagtg gcggtaatac ggttatccac agaatcagaa gggcgaattc cagcacactg
     2881 gcggccgtta ctagtggatc cgagctcggt accaagcttg gcgagatttt caggagctaa
     2941 ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca
     3001 tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt
     3061 tcagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3121 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3181 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3241 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3301 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3361 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3421 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3481 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3541 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3601 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3661 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3721 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3781 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3841 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3901 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3961 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4021 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4081 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4141 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4201 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4261 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4321 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4381 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4441 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4501 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4561 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4621 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg
     4681 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4741 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4801 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4861 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4921 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4981 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5041 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     5101 tatcacgagg ccctttcgtc ttcactcgag gtcgagggat ccagacatga taagatacat
     5161 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     5221 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     5281 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     5341 gtaaaacctc tacaaatgtg gtatggctga ttatgatctc tagtcaaggc actatacatc
     5401 aaatattcct tattaacccc tttacaaatt aaaaagctaa aggtacacaa tttttgagca
     5461 tagttattaa tagcagacac tctatgcctg tgtggagtaa gaaaaaacag tatgttatga
     5521 ttataactgt tatgcctact tataaaggtt acagaatatt tttccataat tttcttgtat
     5581 agcagtgcag ctttttcctt tgtggtgtaa atagcaaagc aagcaagagt tctattacta
     5641 aacacagcat gactcaaaaa acttagcaat tctgaaggaa agtccttggg gtcttctacc
     5701 tttctcttct tttttggagg agtagaatgt tgagagtcag cagtagcctc atcatcacta
     5761 gatggcattt cttctgagca aaacaggttt tcctcattaa aggcattcca ccactgctcc
     5821 cattcatcag ttccataggt tggaatctaa aatacacaaa caattagaat cagtagttta
     5881 acacattata cacttaaaaa ttttatattt accttagagc tttaaatctc tgtaggtagt
     5941 ttgtccaatt atgtcacacc acagaagtaa ggttccttca caaagatccg gaccaaagcg
     6001 gccatcgtgc ctccccactc ctgcagttcg ggggcatgga tgcgcggata gccgctgctg
     6061 gtttcctgga tgccgacgga tttgcactgc cggtagaact ccgcgaggtc gtccagcctc
     6121 aggcagcagc tgaaccaact cgcgagggga tcgagcccgg ggtgggcgaa gaactccagc
     6181 atgagatccc cgcgctggag gatcatccag ccggcgtccc ggaaaacgat tccgaagccc
     6241 aacctttcat agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct
     6301 tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga
     6361 aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc
     6421 attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc tgatagcggt
     6481 ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt tccaccatga
     6541 tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg ggcatgcgcg
     6601 ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg tccagatcat
     6661 cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt
     6721 ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca
     6781 tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc cccggcactt
     6841 cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag
     6901 gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt tcattcaggg
     6961 caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca
     7021 cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat agcctctcca
     7081 cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga aacgatcctc
     7141 atcctgtctc ttgatcagat cttgatcccc tgcgccatca gatccttggc ggcaagaaag
     7201 ccatccagtt tactttgcag ggcttcccaa ccttaccaga gggcgcccca gctggcaatt
     7261 ccggttcgct tgctgtccat aaaaccgccc agtctagcta tcgccatgta agcccactgc
     7321 aagctacctg ctttctcttt gcgcttgcgt tttcccttgt ccagatagcc cagtagctga
     7381 cattcatccg gggtcagcac cgtttctgcg gactggcttt ctacgtgttc cgcttccttt
     7441 agcagccctt gcgccctgag tgcttgcggc agcgtgaagc tagctttttg caaaagccta
     7501 ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg
     7561 cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg
     7621 gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat
     7681 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg
     7741 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     7801 accctaactg acacacattc cacagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc
     7861 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     7921 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc
     7981 agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt
     8041 actgagagtg cacctc
//
