LOCUS       sequence-167256-        4573 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For Flpe/FRT-based Supernova, pK036 should be used with pK068 etc..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4573)
  AUTHORS   Luo W, Mizuno H, Iwata R, Nakazawa S, Yasuda K, Itohara S, Iwasato T
  TITLE     Supernova: A Versatile Vector System for Single-Cell Labeling and 
            Gene Function Studies in vivo.
  JOURNAL   Sci Rep. 2016 Oct 24;6:35747. doi: 10.1038/srep35747.
  PUBMED    27775045
REFERENCE   2  (bases 1 to 4573)
  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: "2016-10-24"; volume: "6:35747. doi"; pages: " 
            10.1038/srep35747"
COMMENT     Sequence Label: pK036.TRE-Flpe-WPRE (Supernova)
FEATURES             Location/Qualifiers
     source          1..4573
                     /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"
     primer_bind     291..315
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      388..397
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(394..399,400..420,421..1692)
                     /codon_start=1
                     /product="nuclear-targeted site-specific recombinase"
                     /label=FLPe
                     /note="FLPe is a fusion between the SV40 nuclear 
                     localization signal and a thermostable version of the 
                     Saccharomyces cerevisiae site-specific recombinase FLP. 
                     Recombination occurs at FRT sequences (Buchholz et al., 
                     1998)."
                     /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA
                     ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK
                     LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI
                     WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV
                     IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY
                     QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS
                     DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL
                     KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
     primer_bind     complement(1805..1825)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2223..2234)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2487..2568
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2524..2543)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2579..2596)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2750..3338)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2830..2849)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3509..4300,4301..4369))
                     /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     4132..4151
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4370..4474)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4542..4560
                     /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 gagaattcga gctcggtacc cggggatcct tgtgctgtct
      361 catcattttg gcaaagaatt acccgccgcc accatggctc ccaagaagaa gaggaaggtg
      421 atgagtcaat ttgatatatt atgtaaaaca ccacctaagg tcctggttcg tcagtttgtg
      481 gaaaggtttg aaagaccttc aggggaaaaa atagcatcat gtgctgctga actaacctat
      541 ttatgttgga tgattactca taacggaaca gcaatcaaga gagccacatt catgagctat
      601 aatactatca taagcaattc gctgagtttc gatattgtca acaaatcact ccagtttaaa
      661 tacaagacgc aaaaagcaac aattctggaa gcctcattaa agaaattaat tcctgcttgg
      721 gaatttacaa ttattcctta caatggacaa aaacatcaat ctgatatcac tgatattgta
      781 agtagtttgc aattacagtt cgaatcatcg gaagaagcag ataagggaaa tagccacagt
      841 aaaaaaatgc ttaaagcact tctaagtgag ggtgaaagca tctgggagat cactgagaaa
      901 atactaaatt cgtttgagta tacctcgaga tttacaaaaa caaaaacttt ataccaattc
      961 ctcttcctag ctactttcat caattgtgga agattcagcg atattaagaa cgttgatccg
     1021 aaatcattta aattagtcca aaataagtat ctgggagtaa taatccagtg tttagtgaca
     1081 gagacaaaga caagcgttag taggcacata tacttcttta gcgcaagggg taggatcgat
     1141 ccacttgtat atttggatga atttttgagg aattctgaac cagtcctaaa acgagtaaat
     1201 aggaccggca attcttcaag caacaaacag gaataccaat tattaaaaga taacttagtc
     1261 agatcgtaca acaaggcttt gaagaaaaat gcgccttatc caatctttgc tataaagaat
     1321 ggcccaaaat ctcacattgg aagacatttg atgacctcat ttctgtcaat gaagggccta
     1381 acggagttga ctaatgttgt gggaaattgg agcgataagc gtgcttctgc cgtggccagg
     1441 acaacgtata ctcatcagat aacagcaata cctgatcact acttcgcact agtttctcgg
     1501 tactatgcat atgatccaat atcaaaggaa atgatagcat tgaaggatga gactaatcca
     1561 attgaggagt ggcagcatat agaacagcta aagggtagtg ctgaaggaag catacgatac
     1621 cccgcatgga atgggataat atcacaggag gtactagact acctttcatc ctacataaat
     1681 agacgcatat aatgatctag agtcgagtta attaagaatt ccgcccgcgg ccgcgaattc
     1741 gcccttcggc cgctcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     1801 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     1861 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     1921 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     1981 caacccccac tggttggggc attgccacca cctgtcagct cctttccgga actttcgctt
     2041 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     2101 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     2161 cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     2221 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     2281 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     2341 agcggccgca tcgataagct tgtcgacgat atctctagag gatcataatc agccatacca
     2401 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     2461 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     2521 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcct cgagcttcct
     2581 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     2641 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     2701 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2761 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2821 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2881 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2941 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3001 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3061 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3121 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3181 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3241 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3301 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3361 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3421 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3481 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3541 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3601 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     3661 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3721 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3781 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     3841 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3901 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     3961 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4021 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4081 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg
     4141 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     4201 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4261 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4321 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     4381 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4441 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     4501 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     4561 cctttcgtct tca
//
