LOCUS       sequence-468211-        5542 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector of AcGFP1 under control of tetracycline responsive
            element (TRE). Also express Tet-on advenced transactivator (rtTA) 
            under control of minimal CMV promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5542)
  AUTHORS   Tsujioka H, Kunieda T, Katou Y, Shirahige K, Fukazawa T, Kubo T
  TITLE     Interleukin-11 induces and maintains progenitors of different cell 
            lineages during Xenopus tadpole tail regeneration.
  JOURNAL   Nat Commun. 2017 Sep 8;8(1):495. doi: 10.1038/s41467-017-00594-5.
  PUBMED    28887447
REFERENCE   2  (bases 1 to 5542)
  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: "Nat 
            Commun."; date: "2017-09-8"; volume: "8(1):495. doi"; pages: " 
            10.1038/s41467-017-00594-5"
COMMENT     Sequence Label: pTon1(acgfp1)
FEATURES             Location/Qualifiers
     source          1..5542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        4..318
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    12..30
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    48..66
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    83..101
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    119..137
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    155..173
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    190..208
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    226..244
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             328..1047
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    1169..1250
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        1339..1718
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1719..1922
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             join(2025..2642,2643..2648,2649..2684,2685..2687,2688..2723,
                     2724..2726,2727..2762,2763..2771)
                     /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"
     polyA_signal    2781..2915
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2805..2824)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    3456..3537
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3548..3565)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3719..4307)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3799..3818)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4478..5269,5270..5338))
                     /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     5101..5120
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5339..5443)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5511..5529
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagttta ctccctatca gtgatagaga acgtatgtcg agtttactcc ctatcagtga
       61 tagagaacga tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttactc
      121 cctatcagtg atagagaacg tatgtcgagt ttactcccta tcagtgatag agaacgtatg
      181 tcgagtttat ccctatcagt gatagagaac gtatgtcgag tttactccct atcagtgata
      241 gagaacgtat gtcgaggtag gcgtgtacgg tgggaggcct atataagcag agctcgttta
      301 gtgaaccgtc agatcgcctg gagaattatg gtgagcaagg gcgccgagct gttcaccggc
      361 atcgtgccca tcctgatcga gctgaatggc gatgtgaatg gccacaagtt cagcgtgagc
      421 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
      481 ggcaagctgc ctgtgccctg gcccaccctg gtgaccaccc tgagctacgg cgtgcagtgc
      541 ttctcacgct accccgatca catgaagcag cacgacttct tcaagagcgc catgcctgag
      601 ggctacatcc aggagcgcac catcttcttc gaggatgacg gcaactacaa gtcgcgcgcc
      661 gaggtgaagt tcgagggcga taccctggtg aatcgcatcg agctgaccgg caccgatttc
      721 aaggaggatg gcaacatcct gggcaataag atggagtaca actacaacgc ccacaatgtg
      781 tacatcatga ccgacaaggc caagaatggc atcaaggtga acttcaagat ccgccacaac
      841 atcgaggatg gcagcgtgca gctggccgac cactaccagc agaatacccc catcggcgat
      901 ggccctgtgc tgctgcccga taaccactac ctgtccaccc agagcgccct gtccaaggac
      961 cccaacgaga agcgcgatca catgatctac ttcggcttcg tgaccgccgc cgccatcacc
     1021 cacggcatgg atgagctgta caagtgagcg gccgcgactc tagatcataa tcagccatac
     1081 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     1141 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     1201 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     1261 tggtttgtcc aattcgagct cggtacccgg ggatcctcta gtcagctgac gcgtcatgtc
     1321 caacattacc gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
     1381 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     1441 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     1501 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     1561 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     1621 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     1681 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgtggttt tggcagtaca
     1741 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     1801 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     1861 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     1921 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
     1981 gaagacaccg ggaccgatcc agcctccgcg gccccgaatt caccatgtct agactggaca
     2041 agagcaaagt cataaacggc gctctggaat tactcaatgg agtcggtatc gaaggcctga
     2101 cgacaaggaa actcgctcaa aagctgggag ttgagcagcc taccctgtac tggcacgtga
     2161 agaacaagcg ggccctgctc gatgccctgc caatcgagat gctggacagg catcataccc
     2221 acttctgccc cctggaaggc gagtcatggc aagactttct gcggaacaac gccaagtcat
     2281 tccgctgtgc tctcctctca catcgcgacg gggctaaagt gcatctcggc acccgcccaa
     2341 cagagaaaca gtacgaaacc ctggaaaatc agctcgcgtt cctgtgtcag caaggcttct
     2401 ccctggagaa cgcactgtac gctctgtccg ccgtgggcca ctttacactg ggctgcgtat
     2461 tggaggaaca ggagcatcaa gtagcaaaag aggaaagaga gacacctacc accgattcta
     2521 tgcccccact tctgagacaa gcaattgagc tgttcgaccg gcagggagcc gaacctgcct
     2581 tccttttcgg cctggaacta atcatatgtg gcctggagaa acagctaaag tgcgaaagcg
     2641 gcgggccggc cgacgccctt gacgattttg acttagacat gctcccagcc gatgcccttg
     2701 acgactttga ccttgatatg ctgcctgctg acgctcttga cgattttgac cttgacatgc
     2761 tccccgggta actaagtaag gatccagaca tgataagata cattgatgag tttggacaaa
     2821 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     2881 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     2941 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     3001 gtggtatggc tgattatgat cctgcaagcc tcgtcgtcct ggccggacca cgctatctgt
     3061 gcaaggtccc cggccccgga cgcgcgctcc atgagcagag cgcccgccgc cgaggcgaag
     3121 actcgggcgg cgccctgccc gtcccaccag gtcaacaggc ggtaaccggc ctcttcatcg
     3181 ggaatgcgcg cgaccttcag catcgccggc atgtccccct ggcggacggg aagtatccag
     3241 ctcgaccaag ctggctccat gtcttccgtc caaggagctt ggcgagattt tcaggagctc
     3301 gcgtgctagc gcggccgcat cgataagctt gtcgacgata tctctagagg atcataatca
     3361 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     3421 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     3481 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcctc
     3541 gagcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3601 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3661 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3721 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3781 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3841 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3901 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3961 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4021 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4081 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4141 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4201 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4261 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4321 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4381 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4441 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4501 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4561 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4621 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4681 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4741 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4801 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4861 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4921 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4981 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5041 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaacacggg
     5101 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5161 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5221 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5281 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5341 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5401 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5461 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     5521 tcacgaggcc ctttcgtctt ca
//
