LOCUS       sequence-297943-        4203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector based on the LiOn integration-coupled translational switch 
            (Kumamoto et al bioRxiv 2019) expressing a functional Cre 
            recombinase from a CMV promoter upon action of the piggyBac 
            transposase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4203)
  AUTHORS   Kumamoto T, Maurinot F, Barry-Martinet R, Vaslin C, 
            Vandormael-Pournin S, Le M, Lerat M, Niculescu D, Cohen-Tannoudji M,
            Rebsam A, Loulier K, Nedelec S, Tozer S, Livet J
  TITLE     Direct Readout of Neural Stem Cell Transgenesis with an 
            Integration-Coupled Gene Expression Switch.
  JOURNAL   Neuron. 2020 Aug 19;107(4):617-630.e6. doi: 
            10.1016/j.neuron.2020.05.038. Epub 2020 Jun 18.
  PUBMED    32559415
REFERENCE   2  (bases 1 to 4203)
  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"; doi: 
            "10.1016/j.neuron.2020.05.038"; journalName: "Neuron"; date: 
            "2020-08-19- 19"; volume: "107"; issue: "4"; pages: "617-630.e6"
COMMENT     Sequence Label: LiOn-CMV∞Cre
FEATURES             Location/Qualifiers
     source          1..4203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        39..342
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        349..552
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     503..523
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             589..609
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     repeat_region   894..928
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    1171..1292
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1208..1227)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1262..1281
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1308..1331)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     repeat_region   2078..2140
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    2252..2363
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2448..3036)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2528..2547)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3207..3998,3999..4067))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3830..3849
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4068..4172)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccaatgatat cggatttaaa tccaagcttg ccgtcgaccg ttacataact tacggtaaat
       61 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      121 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      181 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      241 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      301 acttggcagt acatctacgt attagtcatc gctattacca tgcacgtggt gatgcggttt
      361 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      421 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      481 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      541 ataagcagag ctcgaattcg gatttaaatc gtttaaacca ccatggtgcc caagaagaag
      601 aggaaagtct ccaacctgct gactgtgcac caaaacctgc ctgccctccc tgtggatgcc
      661 acctctgatg aagtcaggaa gaacctgatg gacatgttca gggacaggca ggccttctct
      721 gaacacacct ggaagatgct cctgtctgtg tgcagatcct gggctgcctg gtgcaagctg
      781 aacaacagga aatggttccc tgctgaacct gaggatgtga gggactacct cctgtacctg
      841 caagccagag gcctggctgt gaagaccatc caacagcacc tgggccagct taaccctaga
      901 aagatagtct gcgtaaaatt gacgcatgca ttaagccatg gcttcccgcc ggcggtggcg
      961 gcgcaggatg atggcacgct gcccatgtct tgtgcccagg agagcgggat ggaccgtcac
     1021 cctgcagcct gtgcttctgc taggatcaat gtgtaggcgg ccgcgatcat aatcagccat
     1081 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     1141 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     1201 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     1261 tgtggtttgt ccaaactcat caatgtatct tatcgcgagg atccctactt gtcgtcatcg
     1321 tctttgtagt cgtccccatc ctcgagcagc ctcaccatgg ccccagtctc agagtccagg
     1381 tttctgatgt agttcatcac tatgttcaca ttggtccagc caccagcctg catgatttca
     1441 gggatggaca caccagccct ggccatgtcc ctggcagcac ccactctggc agagtggcca
     1501 gaccaggcca ggtatctctg cccagagtca tccttggcac catagatcag gcggtgggtg
     1561 gcctcaaaga tcccttccag ggcccgggtg gacagttggg aggtggcaga aggggcagcc
     1621 acaccattct ttctgacccg gcagaacagg tagttgttgg ggtcatcagc cacaccagac
     1681 acagagatcc atctctccac cagcttggta acccccaggg acagggcctt ctccacacca
     1741 gctgtggaca ccagggtctt ggtcctgcca atgtggatca gcattctccc accatcggtg
     1801 cgggagatgt ccttcactct gattctggca atttcggcaa tgcgcagcag ggtgttgtag
     1861 gcaatgccca ggaaggccag gttcctgatg tcctggcatc tgtcagagtt ctccatcagg
     1921 gatctgactt ggtcaaagtc agtgcgttca aaggccaggg cctgcttggc tctctcccca
     1981 gcatccacat tctcctttct gattctcctc atcaccaggg acacagcatt ggagtcagaa
     2041 gggcgaggca ggccagatct cctgtgcagc atgttaaccc tagaaagata atcatattgt
     2101 gacgtacgtt aaagataatc atgcgtaaaa ttgacgcatg tcagctactt aactagtccc
     2161 agcatgcctg ctattgtctt cccaatcctc ccccttgctg tcctgcccca ccccaccccc
     2221 cagaatagaa tgacacctac tcagacaatg cgatgcaatt tcctcatttt attaggaaag
     2281 gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga
     2341 tggctggcaa ctagaaggca cagaccggta attgatatcg gaaagaacat gtgagcaaaa
     2401 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2461 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2521 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2581 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2641 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2701 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2761 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2821 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2881 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2941 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3001 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3061 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3121 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3181 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3241 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3301 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3361 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3421 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3481 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3541 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3601 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3661 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3721 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     3781 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     3841 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     3901 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     3961 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4021 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     4081 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4141 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     4201 gtc
//
