LOCUS       sequence-364341-        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        17..331
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    25..43
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    61..79
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    96..114
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    132..150
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    168..186
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    203..221
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    239..257
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     304..328
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      401..410
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(407..412,413..433,434..1705)
                     /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(1818..1838)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2236..2247)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2500..2581
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2537..2556)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2592..2609)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2763..3351)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2843..2862)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3522..4313,4314..4382))
                     /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     4145..4164
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4383..4487)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4555..4573
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cctttcgtct tcactcgagt ttactcccta tcagtgatag agaacgtatg tcgagtttac
       61 tccctatcag tgatagagaa cgatgtcgag tttactccct atcagtgata gagaacgtat
      121 gtcgagttta ctccctatca gtgatagaga acgtatgtcg agtttactcc ctatcagtga
      181 tagagaacgt atgtcgagtt tatccctatc agtgatagag aacgtatgtc gagtttactc
      241 cctatcagtg atagagaacg tatgtcgagg taggcgtgta cggtgggagg cctatataag
      301 cagagctcgt ttagtgaacc gtcagatcgc ctggagaatt cgagctcggt acccggggat
      361 ccttgtgctg tctcatcatt ttggcaaaga attacccgcc gccaccatgg ctcccaagaa
      421 gaagaggaag gtgatgagtc aatttgatat attatgtaaa acaccaccta aggtcctggt
      481 tcgtcagttt gtggaaaggt ttgaaagacc ttcaggggaa aaaatagcat catgtgctgc
      541 tgaactaacc tatttatgtt ggatgattac tcataacgga acagcaatca agagagccac
      601 attcatgagc tataatacta tcataagcaa ttcgctgagt ttcgatattg tcaacaaatc
      661 actccagttt aaatacaaga cgcaaaaagc aacaattctg gaagcctcat taaagaaatt
      721 aattcctgct tgggaattta caattattcc ttacaatgga caaaaacatc aatctgatat
      781 cactgatatt gtaagtagtt tgcaattaca gttcgaatca tcggaagaag cagataaggg
      841 aaatagccac agtaaaaaaa tgcttaaagc acttctaagt gagggtgaaa gcatctggga
      901 gatcactgag aaaatactaa attcgtttga gtatacctcg agatttacaa aaacaaaaac
      961 tttataccaa ttcctcttcc tagctacttt catcaattgt ggaagattca gcgatattaa
     1021 gaacgttgat ccgaaatcat ttaaattagt ccaaaataag tatctgggag taataatcca
     1081 gtgtttagtg acagagacaa agacaagcgt tagtaggcac atatacttct ttagcgcaag
     1141 gggtaggatc gatccacttg tatatttgga tgaatttttg aggaattctg aaccagtcct
     1201 aaaacgagta aataggaccg gcaattcttc aagcaacaaa caggaatacc aattattaaa
     1261 agataactta gtcagatcgt acaacaaggc tttgaagaaa aatgcgcctt atccaatctt
     1321 tgctataaag aatggcccaa aatctcacat tggaagacat ttgatgacct catttctgtc
     1381 aatgaagggc ctaacggagt tgactaatgt tgtgggaaat tggagcgata agcgtgcttc
     1441 tgccgtggcc aggacaacgt atactcatca gataacagca atacctgatc actacttcgc
     1501 actagtttct cggtactatg catatgatcc aatatcaaag gaaatgatag cattgaagga
     1561 tgagactaat ccaattgagg agtggcagca tatagaacag ctaaagggta gtgctgaagg
     1621 aagcatacga taccccgcat ggaatgggat aatatcacag gaggtactag actacctttc
     1681 atcctacata aatagacgca tataatgatc tagagtcgag ttaattaaga attccgcccg
     1741 cggccgcgaa ttcgcccttc ggccgctcaa cctctggatt acaaaatttg tgaaagattg
     1801 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     1861 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     1921 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     1981 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     2041 ggaactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     2101 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     2161 tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc
     2221 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     2281 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     2341 gccgcctccc cgcagcggcc gcatcgataa gcttgtcgac gatatctcta gaggatcata
     2401 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     2461 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     2521 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     2581 cctcgagctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     2641 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     2701 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2761 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2821 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2881 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2941 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3001 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3061 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3121 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3181 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     3241 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3301 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3361 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3421 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3481 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     3541 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     3601 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     3661 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     3721 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     3781 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     3841 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3901 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     3961 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     4021 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     4081 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca
     4141 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     4201 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     4261 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     4321 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     4381 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     4441 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     4501 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     4561 cgtatcacga ggc
//
