LOCUS       sequence-177252-        5317 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5317)
  AUTHORS   Chen F, LoTurco J
  TITLE     A method for stable transgenesis of radial glia lineage in rat 
            neocortex by piggyBac mediated transposition.
  JOURNAL   J Neurosci Methods. 2012 Jun 15;207(2):172-80. Epub 2012 Apr 11.
  PUBMED    22521325
REFERENCE   2  (bases 1 to 5317)
  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: "J Neurosci 
            Methods."; date: "2012-06-15"; volume: "207(2)"; pages: "172-80. 
            Epub 2012 Apr 11"
COMMENT     Sequence Label: PBCAG-Cerulean
FEATURES             Location/Qualifiers
     source          1..5317
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     39..61
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     123..142
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(204..237)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     regulatory      251..260
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(257..259,260..262,263..976)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(302..323)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(563..582)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     910..931
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1063..1082)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1128..1183
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1182..1201)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   1854..1888
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(1911..1927)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1911..1927)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1924..1946)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1935..1951
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1959..1965,1966..1983,1984..1989))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2004..2025
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2084..2279
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2130..2265
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2285..2419
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2322..2341)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2376..2395
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2487..2504)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2658..3246)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2738..2757)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3417..4208,4209..4277))
                     /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     4040..4059
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4278..4382)
                     /gene="bla"
                     /label=AmpR promoter
     repeat_region   4420..4482
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    4779..4812
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        4908..5211
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac
       61 cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct
      121 gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaattcc ctgtaggaat
      181 tggccgcgac tctaggatct aggataactt cgtataggat actttatacg aagttattga
      241 tccaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
      301 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
      361 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
      421 cgtgccctgg cccaccctcg tgaccaccct gacctggggc gtgcagtgct tcgcccgcta
      481 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
      541 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
      601 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
      661 caacatcctg gggcacaagc tggagtacaa cgccatcagc gacaacgtct atatcaccgc
      721 cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg
      781 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
      841 gctgcccgac aaccactacc tgagcaccca gtccaagctg agcaaagacc ccaacgagaa
      901 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
      961 cgagctgtac aagtaaagcg gccgcactcc tcaggtgcag gctgcctatc agaaggtggt
     1021 ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa
     1081 aaattatggg gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt
     1141 tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg
     1201 gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc
     1261 ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca
     1321 gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt
     1381 tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt
     1441 actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt
     1501 atggagatcc ctcgacctgc agagctccag cctacacaat cgctcaagac gtgtaatgct
     1561 tttattatat attagtcacg atatctataa caagaaaata tatatataat aagttatcac
     1621 gtaagtagaa catgaaataa caatataatt atcgtatgag ttaaatctta aaagtcacgt
     1681 aaaagataat catgcgtcat tttgactcac gcggtcgtta tagttcaaaa tcagtgacac
     1741 ttaccgcatt gacaagcacg cctcacggga gctccaagcg gcgactgaga tgtcctaaat
     1801 gcacagcgac ggattcgcgc tatttagaaa gagagagcaa tatttcaaga atgcatgcgt
     1861 caattttacg cagactatct ttctagggtt aaaagcttgg cgtaatcatg gtcatagctg
     1921 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     1981 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     2041 ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga tccgcatctc aataagtcag
     2101 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     2161 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     2221 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     2281 agctaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     2341 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     2401 atcttatcat gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     2461 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     2521 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2581 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2641 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2701 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2761 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2821 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2881 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2941 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3001 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3061 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3121 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3181 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3241 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3301 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3361 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3421 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3481 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3541 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3601 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3661 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     3721 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     3781 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     3841 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3901 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3961 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     4021 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     4081 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     4141 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     4201 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     4261 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     4321 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     4381 cgcacatttc cccgaaaagt gccacctggg tcgacttaac cctagaaaga taatcatatt
     4441 gtgacgtacg ttaaagataa tcatgcgtaa aattgacgca tgtgttttat cgatctgtat
     4501 atcgaggttt atttattaat ttgaatagat attaagtttt attatattta cacttacata
     4561 ctaataataa attcaacaaa caatttattt atgtttattt atttattaaa aaaaaacaaa
     4621 aactcaaaat ttcttctata aagtaacaaa acttttaaac attctctctt ttacaaaaat
     4681 aaacttattt tgtactttaa aaacagtcat gttgtattat aaaataagta attagcttaa
     4741 cttatacata atagaaacaa attatactta ttaatcgcat aacttcgtat aatgtatgct
     4801 atacgaagtt atgcgattaa gggatctgta gggcgcagta gtccaactag ttattaatag
     4861 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     4921 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     4981 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     5041 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     5101 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     5161 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg
     5221 agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat
     5281 ttatttattt tttaattatt ttgtgcagcg atggggg
//
