LOCUS       sequence-468199-        6194 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector of X. laevis il11.L-P2A-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 6194)
  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 6194)
  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(il11)
FEATURES             Location/Qualifiers
     source          1..6194
                     /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             923..979
                     /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"
     CDS             980..1699
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    1821..1902
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        1991..2370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2371..2574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             join(2677..3294,3295..3300,3301..3336,3337..3339,3340..3375,
                     3376..3378,3379..3414,3415..3423)
                     /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    3433..3567
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3457..3476)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    4108..4189
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4200..4217)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4371..4959)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4451..4470)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5130..5921,5922..5990))
                     /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     5753..5772
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5991..6095)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6163..6181
                     /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 gagaattccc caaagagcca aaacaaaata tgaaaagcag
      361 tttgtgccag atcgctctta cagtactgag cctttgtgag ggactacggg ctgttccctt
      421 gaaaccaaaa gtggaatttg atgaccttct aaacatggct atgcaccttc taagagatac
      481 caaacaatca taccacaact ttaaagccaa atatccatca gaaggagagc acaaactgga
      541 cagcctgcct gtagtctcaa tgaatgctgt agaaatctct aatgtacaga tacacacaag
      601 cctcctgaag ctaagcactg acctctttat atatcagaag cattttgact ggctgagaaa
      661 ggcagcacat gccattcgcc ccatggacca cgaatttaac ggaattcaca atcgcattga
      721 aaagctgttg aaaaaaattg atctcctgat gatcaggcac catatgccac gtgtcacgga
      781 acccgttgct ccacagttat ctaactcaac cactccatgg ggggtcattc agagcggtca
      841 tgccattttc cgtcatttcc acctcttctt ggactatgca acacgtgttc tggttctgat
      901 gaagaacaag ttgggaagcg gagctactaa cttcagcctg ctgaagcagg ctggagacgt
      961 ggaggagaac cctggaccta tggtgagcaa gggcgccgag ctgttcaccg gcatcgtgcc
     1021 catcctgatc gagctgaatg gcgatgtgaa tggccacaag ttcagcgtga gcggcgaggg
     1081 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     1141 gcctgtgccc tggcccaccc tggtgaccac cctgagctac ggcgtgcagt gcttctcacg
     1201 ctaccccgat cacatgaagc agcacgactt cttcaagagc gccatgcctg agggctacat
     1261 ccaggagcgc accatcttct tcgaggatga cggcaactac aagtcgcgcg ccgaggtgaa
     1321 gttcgagggc gataccctgg tgaatcgcat cgagctgacc ggcaccgatt tcaaggagga
     1381 tggcaacatc ctgggcaata agatggagta caactacaac gcccacaatg tgtacatcat
     1441 gaccgacaag gccaagaatg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     1501 tggcagcgtg cagctggccg accactacca gcagaatacc cccatcggcg atggccctgt
     1561 gctgctgccc gataaccact acctgtccac ccagagcgcc ctgtccaagg accccaacga
     1621 gaagcgcgat cacatgatct acttcggctt cgtgaccgcc gccgccatca cccacggcat
     1681 ggatgagctg tacaagtgag cggccgcgac tctagatcat aatcagccat accacatttg
     1741 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     1801 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     1861 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     1921 ccaattcgag ctcggtaccc ggggatcctc tagtcagctg acgcgtcatg tccaacatta
     1981 ccgccatgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     2041 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     2101 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     2161 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     2221 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     2281 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     2341 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2401 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2461 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2521 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
     2581 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
     2641 cgggaccgat ccagcctccg cggccccgaa ttcaccatgt ctagactgga caagagcaaa
     2701 gtcataaacg gcgctctgga attactcaat ggagtcggta tcgaaggcct gacgacaagg
     2761 aaactcgctc aaaagctggg agttgagcag cctaccctgt actggcacgt gaagaacaag
     2821 cgggccctgc tcgatgccct gccaatcgag atgctggaca ggcatcatac ccacttctgc
     2881 cccctggaag gcgagtcatg gcaagacttt ctgcggaaca acgccaagtc attccgctgt
     2941 gctctcctct cacatcgcga cggggctaaa gtgcatctcg gcacccgccc aacagagaaa
     3001 cagtacgaaa ccctggaaaa tcagctcgcg ttcctgtgtc agcaaggctt ctccctggag
     3061 aacgcactgt acgctctgtc cgccgtgggc cactttacac tgggctgcgt attggaggaa
     3121 caggagcatc aagtagcaaa agaggaaaga gagacaccta ccaccgattc tatgccccca
     3181 cttctgagac aagcaattga gctgttcgac cggcagggag ccgaacctgc cttccttttc
     3241 ggcctggaac taatcatatg tggcctggag aaacagctaa agtgcgaaag cggcgggccg
     3301 gccgacgccc ttgacgattt tgacttagac atgctcccag ccgatgccct tgacgacttt
     3361 gaccttgata tgctgcctgc tgacgctctt gacgattttg accttgacat gctccccggg
     3421 taactaagta aggatccaga catgataaga tacattgatg agtttggaca aaccacaact
     3481 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     3541 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     3601 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg
     3661 gctgattatg atcctgcaag cctcgtcgtc ctggccggac cacgctatct gtgcaaggtc
     3721 cccggccccg gacgcgcgct ccatgagcag agcgcccgcc gccgaggcga agactcgggc
     3781 ggcgccctgc ccgtcccacc aggtcaacag gcggtaaccg gcctcttcat cgggaatgcg
     3841 cgcgaccttc agcatcgccg gcatgtcccc ctggcggacg ggaagtatcc agctcgacca
     3901 agctggctcc atgtcttccg tccaaggagc ttggcgagat tttcaggagc tcgcgtgcta
     3961 gcgcggccgc atcgataagc ttgtcgacga tatctctaga ggatcataat cagccatacc
     4021 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     4081 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     4141 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgcc tcgagcttcc
     4201 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4261 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4321 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4381 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4441 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4501 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4561 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4621 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4681 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4741 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4801 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4861 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4921 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4981 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5041 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5101 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5161 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5221 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5281 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5341 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5401 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5461 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5521 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5581 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5641 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5701 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc
     5761 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5821 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5881 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5941 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6001 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6061 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6121 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     6181 ccctttcgtc ttca
//
