LOCUS       sequence-177250-        5111 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 5111)
  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 5111)
  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-mRFP
FEATURES             Location/Qualifiers
     source          1..5111
                     /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"
     CDS             170..847
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRPS
                     TGA"
     primer_bind     complement(300..318)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     568..587
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     762..785
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(851..870)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    916..971
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(970..989)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   1642..1676
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(1699..1715)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1699..1715)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1712..1734)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1723..1739
                     /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(1747..1753,1754..1771,1772..1777))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1792..1813
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1872..2067
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1918..2053
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2073..2207
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2110..2129)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2164..2183
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2275..2292)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2446..3034)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2526..2545)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3205..3996,3997..4065))
                     /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     3828..3847
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4066..4170)
                     /gene="bla"
                     /label=AmpR promoter
     repeat_region   4208..4270
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    4567..4600
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        4696..4999
                     /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 aaagaattca tggcctcctc
      181 cgaggacgtc atcaaggagt tcatgcgctt caaggtgcgc atggagggct ccgtgaacgg
      241 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
      301 caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccctca
      361 gttccagtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa
      421 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg atggcggcgt
      481 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
      541 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga
      601 ggcctccacc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagatgag
      661 gctgaagctg aaggacggcg gccactacga cgccgaggtc aagaccacct acatggccaa
      721 gaagcccgtg cagctgcccg gcgcctacaa gaccgacatc aagctggaca tcacctccca
      781 caacgaggac tacaccatcg tggaacagta cgagcgcgcc gagggccgcc cctccaccgg
      841 cgcctaaaga tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc
      901 atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt
      961 tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt
     1021 atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc
     1081 tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa
     1141 aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa
     1201 catccctaaa attttcctta catgttttac tagccagatt tttcctcctc tcctgactac
     1261 tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcag agctccagcc
     1321 tacacaatcg ctcaagacgt gtaatgcttt tattatatat tagtcacgat atctataaca
     1381 agaaaatata tatataataa gttatcacgt aagtagaaca tgaaataaca atataattat
     1441 cgtatgagtt aaatcttaaa agtcacgtaa aagataatca tgcgtcattt tgactcacgc
     1501 ggtcgttata gttcaaaatc agtgacactt accgcattga caagcacgcc tcacgggagc
     1561 tccaagcggc gactgagatg tcctaaatgc acagcgacgg attcgcgcta tttagaaaga
     1621 gagagcaata tttcaagaat gcatgcgtca attttacgca gactatcttt ctagggttaa
     1681 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     1741 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     1801 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     1861 ccagcggatc cgcatctcaa taagtcagca accatagtcc cgcccctaac tccgcccatc
     1921 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     1981 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     2041 ttttttggag gcctaggctt ttgcaaaaag ctaacttgtt tattgcagct tataatggtt
     2101 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     2161 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatccgc tgcattaatg
     2221 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     2281 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2341 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2401 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2461 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2521 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2581 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2641 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2701 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2761 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2821 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2881 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2941 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3001 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3061 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3121 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3181 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3241 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3301 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3361 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3421 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3481 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3541 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3601 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3661 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     3721 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     3781 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     3841 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     3901 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     3961 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4021 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4081 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4141 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgggtc
     4201 gacttaaccc tagaaagata atcatattgt gacgtacgtt aaagataatc atgcgtaaaa
     4261 ttgacgcatg tgttttatcg atctgtatat cgaggtttat ttattaattt gaatagatat
     4321 taagttttat tatatttaca cttacatact aataataaat tcaacaaaca atttatttat
     4381 gtttatttat ttattaaaaa aaaacaaaaa ctcaaaattt cttctataaa gtaacaaaac
     4441 ttttaaacat tctctctttt acaaaaataa acttattttg tactttaaaa acagtcatgt
     4501 tgtattataa aataagtaat tagcttaact tatacataat agaaacaaat tatacttatt
     4561 aatcgcataa cttcgtataa tgtatgctat acgaagttat gcgattaagg gatctgtagg
     4621 gcgcagtagt ccaactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     4681 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     4741 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     4801 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     4861 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     4921 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     4981 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
     5041 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
     5101 gggggcgggg g
//
