LOCUS       sequence-308112-        5467 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector based on the LiOn integration-coupled translational switch 
            (Kumamoto et al bioRxiv 2019) expressing the fluorescent protein 
            IRFP670 from a CAG promoter upon action of the piggyBac transposase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5467)
  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 5467)
  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-CAG∞IRFP
FEATURES             Location/Qualifiers
     source          1..5467
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        383..658
                     /label=chicken beta-actin promoter
     intron          659..1667
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1591..1613
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1675..1694
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1732..1741
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   1749..1783
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     repeat_region   2987..3049
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     3405..3424
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    3423..3478
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     3524..3543
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     rep_origin      complement(3690..4278)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3770..3789)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4449..5240,5241..5309))
                     /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     5072..5091
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5310..5414)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggactatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg ggtcgaggtg agccccacgt tctgcttcac tctccccatc
      421 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg
      481 atgggggcgg gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg
      541 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
      601 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
      661 agtcgctgcg ttgccttcgc cccgtgcccc gctccgcgcc gcctcgcgcc gcccgccccg
      721 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg
      781 ctgtaattag cgcttggttt aatgacggct cgtttctttt ctgtggctgc gtgaaagcct
      841 taaagggctc cgggagggcc ctttgtgcgg gggggagcgg ctcggggggt gcgtgcgtgt
      901 gtgtgtgcgt ggggagcgcc gcgtgcggcc cgcgctgccc ggcggctgtg agcgctgcgg
      961 gcgcggcgcg gggctttgtg cgctccgcgt gtgcgcgagg ggagcgcggc cgggggcggt
     1021 gccccgcggt gcgggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg
     1081 gggggtgagc agggggtgtg ggcgcggcgg tcgggctgta acccccccct gcacccccct
     1141 ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtgcggg gcgtggcgcg
     1201 gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg
     1261 cctcgggccg gggagggctc gggggagggg cgcggcggcc ccggagcgcc ggcggctgtc
     1321 gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac
     1381 ttcctttgtc ccaaatctgg cggagccgaa atctgggagg cgccgccgca ccccctctag
     1441 cgggcgcggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt
     1501 gcgtcgccgc gccgccgtcc ccttctccat ctccagcctc ggggctgccg cagggggacg
     1561 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
     1621 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
     1681 tgctggttat tgtgctgtct catcattttg gcaaagcacg tgaggaccgg tgccaccatg
     1741 gcgtttaacc ctagaaagat agtctgcgta aaattgacgc atgcatcaat tgtaactgcg
     1801 gtttaaactc cccagcatgc ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc
     1861 ccaccccacc ccccagaata gaatgacacc tactcagaca atgcgatgca atttcctcat
     1921 tttattagga aaggacagtg agagtggcac cttccagggt caaggaaggc acgggggagg
     1981 ggcaaacaac agatggctgg caactagaag gcactaaagt cggatccgaa ttcattcccg
     2041 ggtagagtcg cggccgctta gcgttggtgg tgggcggcgg tgaagtgcag cgataagaag
     2101 tcggcgaaca tttcggccgc gacgcgctgc gccatcggca cggcacgcgg ctcgtaatga
     2161 tgacagatga tcaatcccca tagcgtgccg tcaatgatga tcgacagcga catcgaggcg
     2221 ctgacgccca tgttccgcag aaattcgaga tggcagggcg agatgctgcg caggtgcgcg
     2281 aacgacagat cgagcgcggc gccggaggcg tcgtgctcgg gcacgatccg gctgctgatg
     2341 ccgcgcgaat ccgagaccac gcggatcgcg ttcttcaagt acagtagccg cgcctgctgc
     2401 gggaccagcg acgccggaaa gtgctgaccg agaaagctct cgaggtcgct gcgcttcgcc
     2461 tcgccgatca ccatcccgga gccgtcgtcc gcgaagcggt acaacatcac gcggtgatag
     2521 ccgagcatcg cctgcagata gcgcggcacc cgtgcggcca tctcttcgag cgacttcagt
     2581 tctttggtgc gcgcgatgat ctgccgcgtc agccgcagcg gattgtcggc ctgttcggcc
     2641 gccgcaggct cgaactcgat gatcgatgta ccgtcatggc gatgcagtga gatgtcgaag
     2701 gtgcggccgg tcaggccgtc gcgccaaccg aagatcagcg ccggtcgctt tggatcggag
     2761 gactgcgcca gtgcgttgcg cagcgcatgg gcttcggtct cgccgaagta atcggcgagt
     2821 agctcaccga cccgcggagt ttcgcgtcca aagaacgcgc cggcattttc cgtaatgcgc
     2881 gtgatccgca ccgcctgcgc gtcgcaggct agcaggcagc cgcacggctg aatgctgccg
     2941 gggatgtgga tcggctcgcg atcgcaggag gtgagatcga cattaaccct agaaagataa
     3001 tcatattgtg acgtacgtta aagataatca tgcgtaaaat tgacgcatgt ccggagccgt
     3061 agatatcgga aagaaagcgg ccgctgcagg tcgagggatc tccataagag aagagggaca
     3121 gctatgactg ggagtagtca ggagaggagg aaaaatctgg ctagtaaaac atgtaaggaa
     3181 aattttaggg atgttaaaga aaaaaataac acaaaacaaa atataaaaaa aatctaacct
     3241 caagtcaagg cttttctatg gaataaggaa tggacagcag ggggctgttt catatactga
     3301 tgacctcttt atagccaacc tttgttcatg gcagccagca tatgggcata tgttgccaaa
     3361 ctctaaacca aatactcatt ctgatgtttt aaatgatttg ccctcccata tgtccttccg
     3421 agtgagagac acaaaaaatt ccaacacact attgcaatga aaataaattt cctttattag
     3481 ccagaagtca gatgctcaag gggcttcatg atgtccccat aatttttggc agagggaaaa
     3541 agatctcagt ggtatttgtg agccagggca ttggccacac cagccaccac cttctgatag
     3601 gcagcctgca cctgaggagt gaattagaag ctttgagcaa aaggccagca aaaggccagg
     3661 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3721 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3781 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3841 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3901 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3961 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4021 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4081 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4141 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4201 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4261 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4321 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4381 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4441 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4501 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4561 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4621 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4681 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4741 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4801 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4861 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4921 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4981 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5041 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5101 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5161 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5221 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5281 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5341 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5401 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcccaat gatatcattt
     5461 aaatgtc
//
