LOCUS       sequence-177240-        8169 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 8169)
  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 8169)
  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-EGFR
FEATURES             Location/Qualifiers
     source          1..8169
                     /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             join(186..257,258..2120,2121..2189,2190..3818)
                     /codon_start=1
                     /product="precursor of human epidermal growth factor 
                     receptor"
                     /label=EGFR
                     /translation="MRPSGTAGAALLALLAALCPASRALEEKKVCQGTSNKLTQLGTFE
                     DHFLSLQRMFNNCEVVLGNLEITYVQRNYDLSFLKTIQEVAGYVLIALNTVERIPLENL
                     QIIRGNMYYENSYALAVLSNYDANKTGLKELPMRNLQEILHGAVRFSNNPALCNVESIQ
                     WRDIVSSDFLSNMSMDFQNHLGSCQKCDPSCPNGSCWGAGEENCQKLTKIICAQQCSGR
                     CRGKSPSDCCHNQCAAGCTGPRESDCLVCRKFRDEATCKDTCPPLMLYNPTTYQMDVNP
                     EGKYSFGATCVKKCPRNYVVTDHGSCVRACGADSYEMEEDGVRKCKKCEGPCRKVCNGI
                     GIGEFKDSLSINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVK
                     EITGFLLIQAWPENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLNITSLGLRSLKEISD
                     GDVIISGNKNLCYANTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWG
                     PEPRDCVSCRNVSRGRECVDKCNLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGP
                     DNCIQCAHYIDGPHCVKTCPAGVMGENNTLVWKYADAGHVCHLCHPNCTYGCTGPGLEG
                     CPTNGPKISSIATGMVGALLLLLVVALGIGLFMRRRHIVRKRTLRRLLQERELVEPLTP
                     SGEAPNQALLRILKETEFKKIKVLGSGAFGTVYKGLWIPEGEKVKIPVAIKELREATSP
                     KANKEILDEAYVMASVDNPHVCRLLGICLTSTVQLITQLMPFGCLLDYVREHKDNIGSQ
                     YLLNWCVQIAKGMNYLEDRRLVHRDLAARNVLVKTPQHVKITDFGLAKLLGAEEKEYHA
                     EGGKVPIKWMALESILHRIYTHQSDVWSYGVTVWELMTFGSKPYDGIPASEISSILEKG
                     ERLPQPPICTIDVYMIMVKCWMIDADSRPKFRELIIEFSKMARDPQRYLVIQGDERMHL
                     PSPTDSNFYRALMDEEDMDDVVDADEYLIPQQGFFSSPSTSRTPLLSSLSATSNNSTVA
                     CIDRNGLQSCPIKEDSFLQRYSSDPTGALTEDSIDDTFLPVPEYINQSVPKRPAGSVQN
                     PVYHNQPLNPAPSRDPHYQDPHSTAVGNPEYLNTVQPTCVNSTFDSPAHWAQKGSHQIS
                     LDNPDYQQDFFPKEAKPNGIFKGSTAENAEYLRVAPQSSEFIGA"
     primer_bind     complement(3904..3923)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3969..4024
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4023..4042)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   4695..4729
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(4752..4768)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4752..4768)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4765..4787)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4776..4792
                     /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(4800..4806,4807..4824,4825..4830))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4845..4866
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4925..5120
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4971..5106
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5126..5260
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5163..5182)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5217..5236
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5328..5345)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5499..6087)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5579..5598)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6258..7049,7050..7118))
                     /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"
     promoter        complement(7119..7223)
                     /gene="bla"
                     /label=AmpR promoter
     repeat_region   7261..7323
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    7620..7653
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        7749..8052
                     /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 aaagaattct gcagtcgacg
      181 gtaccatgcg accctccggg acggccgggg cagcgctcct ggcgctgctg gctgcgctct
      241 gcccggcgag tcgggctctg gaggaaaaga aagtttgcca aggcacgagt aacaagctca
      301 cgcagttggg cacttttgaa gatcattttc tcagcctcca gaggatgttc aataactgtg
      361 aggtggtcct tgggaatttg gaaattacct atgtgcagag gaattatgat ctttccttct
      421 taaagaccat ccaggaggtg gctggttatg tcctcattgc cctcaacaca gtggagcgaa
      481 ttcctttgga aaacctgcag atcatcagag gaaatatgta ctacgaaaat tcctatgcct
      541 tagcagtctt atctaactat gatgcaaata aaaccggact gaaggagctg cccatgagaa
      601 atttacagga aatcctgcat ggcgccgtgc ggttcagcaa caaccctgcc ctgtgcaacg
      661 tggagagcat ccagtggcgg gacatagtca gcagtgactt tctcagcaac atgtcgatgg
      721 acttccagaa ccacctgggc agctgccaaa agtgtgatcc aagctgtccc aatgggagct
      781 gctggggtgc aggagaggag aactgccaga aactgaccaa aatcatctgt gcccagcagt
      841 gctccgggcg ctgccgtggc aagtccccca gtgactgctg ccacaaccag tgtgctgcag
      901 gctgcacagg cccccgggag agcgactgcc tggtctgccg caaattccga gacgaagcca
      961 cgtgcaagga cacctgcccc ccactcatgc tctacaaccc caccacgtac cagatggatg
     1021 tgaaccccga gggcaaatac agctttggtg ccacctgcgt gaagaagtgt ccccgtaatt
     1081 atgtggtgac agatcacggc tcgtgcgtcc gagcctgtgg ggccgacagc tatgagatgg
     1141 aggaagacgg cgtccgcaag tgtaagaagt gcgaagggcc ttgccgcaaa gtgtgtaacg
     1201 gaataggtat tggtgaattt aaagactcac tctccataaa tgctacgaat attaaacact
     1261 tcaaaaactg cacctccatc agtggcgatc tccacatcct gccggtggca tttaggggtg
     1321 actccttcac acatactcct cctctggatc cacaggaact ggatattctg aaaaccgtaa
     1381 aggaaatcac agggtttttg ctgattcagg cttggcctga aaacaggacg gacctccatg
     1441 cctttgagaa cctagaaatc atacgcggca ggaccaagca acatggtcag ttttctcttg
     1501 cagtcgtcag cctgaacata acatccttgg gattacgctc cctcaaggag ataagtgatg
     1561 gagatgtgat aatttcagga aacaaaaatt tgtgctatgc aaatacaata aactggaaaa
     1621 aactgtttgg gacctccggt cagaaaacca aaattataag caacagaggt gaaaacagct
     1681 gcaaggccac aggccaggtc tgccatgcct tgtgctcccc cgagggctgc tggggcccgg
     1741 agcccaggga ctgcgtctct tgccggaatg tcagccgagg cagggaatgc gtggacaagt
     1801 gcaaccttct ggagggtgag ccaagggagt ttgtggagaa ctctgagtgc atacagtgcc
     1861 acccagagtg cctgcctcag gccatgaaca tcacctgcac aggacgggga ccagacaact
     1921 gtatccagtg tgcccactac attgacggcc cccactgcgt caagacctgc ccggcaggag
     1981 tcatgggaga aaacaacacc ctggtctgga agtacgcaga cgccggccat gtgtgccacc
     2041 tgtgccatcc aaactgcacc tacggatgca ctgggccagg tcttgaaggc tgtccaacga
     2101 atgggcctaa gatctcgtcc atcgccactg ggatggtggg ggccctcctc ttgctgctgg
     2161 tggtggccct ggggatcggc ctcttcatgc gaaggcgcca catcgttcgg aagcgcacgc
     2221 tgcggaggct gctgcaggag agggagcttg tggagcctct tacacccagt ggagaagctc
     2281 ccaaccaagc tctcttgagg atcttgaagg aaactgaatt caaaaagatc aaagtgctgg
     2341 gctccggtgc gttcggcacg gtgtataagg gactctggat cccagaaggt gagaaagtta
     2401 aaattcccgt cgctatcaag gaattaagag aagcaacatc tccgaaggcc aacaaggaaa
     2461 tcctcgatga agcctacgtg atggccagcg tggacaaccc ccacgtgtgc cgcctgctgg
     2521 gcatctgcct cacctccacc gtgcaactca tcacgcagct catgcccttc ggctgcctcc
     2581 tggactatgt ccgggaacac aaagacaata ttggctccca gtacctgctc aactggtgtg
     2641 tgcagatcgc aaagggcatg aactacttgg aggaccgtcg cttggtgcac cgcgacctgg
     2701 cagccaggaa cgtactggtg aaaacaccgc agcatgtcaa gatcacagat tttgggctgg
     2761 ccaaactgct gggtgcggaa gagaaagaat accatgcaga aggaggcaaa gtgcctatca
     2821 agtggatggc attggaatca attttacaca gaatctatac ccaccagagt gatgtctgga
     2881 gctacggggt gaccgtttgg gagttgatga cctttggatc caagccatat gacggaatcc
     2941 ctgccagcga gatctcctcc atcctggaga aaggagaacg cctccctcag ccacccatat
     3001 gtaccatcga tgtctacatg atcatggtca agtgctggat gatagacgca gatagtcgcc
     3061 caaagttccg tgagttgatc atcgaattct ccaaaatggc ccgagacccc cagcgctacc
     3121 ttgtcattca gggggatgaa agaatgcatt tgccaagtcc tacagactcc aacttctacc
     3181 gtgccctgat ggatgaagaa gacatggacg acgtggtgga tgccgacgag tacctcatcc
     3241 cacagcaggg cttcttcagc agcccctcca cgtcacggac tcccctcctg agctctctga
     3301 gtgcaaccag caacaattcc accgtggctt gcattgatag aaatgggctg caaagctgtc
     3361 ccatcaagga agacagcttc ttgcagcgat acagctcaga ccccacaggc gccttgactg
     3421 aggacagcat agacgacacc ttcctcccag tgcctgaata cataaaccag tccgttccca
     3481 aaaggcccgc tggctctgtg cagaatcctg tctatcacaa tcagcctctg aaccccgcgc
     3541 ccagcagaga cccacactac caggaccccc acagcactgc agtgggcaac cccgagtatc
     3601 tcaacactgt ccagcccacc tgtgtcaaca gcacattcga cagccctgcc cactgggccc
     3661 agaaaggcag ccaccaaatt agcctggaca accctgacta ccagcaggac ttctttccca
     3721 aggaagccaa gccaaatggc atctttaagg gctccacagc tgaaaatgca gaatacctaa
     3781 gggtcgcgcc acaaagcagt gaatttattg gagcatgagc ggccgcactc ctcaggtgca
     3841 ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg
     3901 agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac
     3961 ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
     4021 ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt
     4081 ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag
     4141 aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt
     4201 gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct
     4261 aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt
     4321 catagctgtc cctcttctct tatggagatc cctcgacctg cagagctcca gcctacacaa
     4381 tcgctcaaga cgtgtaatgc ttttattata tattagtcac gatatctata acaagaaaat
     4441 atatatataa taagttatca cgtaagtaga acatgaaata acaatataat tatcgtatga
     4501 gttaaatctt aaaagtcacg taaaagataa tcatgcgtca ttttgactca cgcggtcgtt
     4561 atagttcaaa atcagtgaca cttaccgcat tgacaagcac gcctcacggg agctccaagc
     4621 ggcgactgag atgtcctaaa tgcacagcga cggattcgcg ctatttagaa agagagagca
     4681 atatttcaag aatgcatgcg tcaattttac gcagactatc tttctagggt taaaagcttg
     4741 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4801 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4861 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg
     4921 atccgcatct caataagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4981 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     5041 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     5101 gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg gttacaaata
     5161 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5221 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta atgaatcggc
     5281 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5341 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5401 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5461 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5521 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5581 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5641 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5701 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5761 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5821 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5881 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5941 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6001 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6061 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6121 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6181 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6241 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6301 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6361 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6421 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6481 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6541 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6601 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6661 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6721 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6781 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6841 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6901 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6961 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7021 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7081 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7141 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7201 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgg gtcgacttaa
     7261 ccctagaaag ataatcatat tgtgacgtac gttaaagata atcatgcgta aaattgacgc
     7321 atgtgtttta tcgatctgta tatcgaggtt tatttattaa tttgaataga tattaagttt
     7381 tattatattt acacttacat actaataata aattcaacaa acaatttatt tatgtttatt
     7441 tatttattaa aaaaaaacaa aaactcaaaa tttcttctat aaagtaacaa aacttttaaa
     7501 cattctctct tttacaaaaa taaacttatt ttgtacttta aaaacagtca tgttgtatta
     7561 taaaataagt aattagctta acttatacat aatagaaaca aattatactt attaatcgca
     7621 taacttcgta taatgtatgc tatacgaagt tatgcgatta agggatctgt agggcgcagt
     7681 agtccaacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     7741 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     7801 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     7861 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     7921 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     7981 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     8041 gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc
     8101 tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg
     8161 ggggggggg
//
