LOCUS       sequence-418745-        6348 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6348)
  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 6348)
  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-eGFP
FEATURES             Location/Qualifiers
     source          1..6348
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   12..74
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    371..404
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        500..803
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        805..1081
                     /label=chicken beta-actin promoter
     intron          1082..2098
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2022..2044
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2106..2125
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      2191..2200
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2197..2199,2200..2202,2203..2916)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2197..2199,2200..2202,2203..2913)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2242..2263)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2503..2522)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2850..2871
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3003..3022)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3068..3123
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3122..3141)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   3794..3828
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(3851..3867)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3851..3867)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3864..3886)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3875..3891
                     /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(3899..3905,3906..3923,3924..3929))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3944..3965
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4024..4219
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4070..4205
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4225..4359
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4262..4281)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4316..4335
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4427..4444)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4598..5186)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4678..4697)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5357..6148,6149..6217))
                     /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     5980..5999
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6218..6322)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgactta accctagaaa gataatcata ttgtgacgta cgttaaagat aatcatgcgt
       61 aaaattgacg catgtgtttt atcgatctgt atatcgaggt ttatttatta atttgaatag
      121 atattaagtt ttattatatt tacacttaca tactaataat aaattcaaca aacaatttat
      181 ttatgtttat ttatttatta aaaaaaaaca aaaactcaaa atttcttcta taaagtaaca
      241 aaacttttaa acattctctc ttttacaaaa ataaacttat tttgtacttt aaaaacagtc
      301 atgttgtatt ataaaataag taattagctt aacttataca taatagaaac aaattatact
      361 tattaatcgc ataacttcgt ataatgtatg ctatacgaag ttatgcgatt aagggatctg
      421 tagggcgcag tagtccaact agttattaat agtaatcaat tacggggtca ttagttcata
      481 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      541 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      601 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      661 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      721 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      781 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      841 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      901 cgatgggggc gggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      961 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
     1021 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
     1081 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
     1141 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
     1201 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
     1261 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
     1321 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
     1381 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1441 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1501 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1561 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1621 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1681 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1741 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1801 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1861 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1921 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1981 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     2041 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     2101 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tctgcagtcg
     2161 acggtaccgc gggcccggga tccaccggtc gccaccatgg tgagcaaggg cgaggagctg
     2221 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     2281 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     2341 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     2401 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     2461 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     2521 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     2581 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     2641 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     2701 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     2761 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     2821 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     2881 gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgcactcc tcaggtgcag
     2941 gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga
     3001 gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact
     3061 tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc
     3121 tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt
     3181 tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga
     3241 ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg
     3301 acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta
     3361 aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc
     3421 atagctgtcc ctcttctctt atggagatcc ctcgacctgc agagctccag cctacacaat
     3481 cgctcaagac gtgtaatgct tttattatat attagtcacg atatctataa caagaaaata
     3541 tatatataat aagttatcac gtaagtagaa catgaaataa caatataatt atcgtatgag
     3601 ttaaatctta aaagtcacgt aaaagataat catgcgtcat tttgactcac gcggtcgtta
     3661 tagttcaaaa tcagtgacac ttaccgcatt gacaagcacg cctcacggga gctccaagcg
     3721 gcgactgaga tgtcctaaat gcacagcgac ggattcgcgc tatttagaaa gagagagcaa
     3781 tatttcaaga atgcatgcgt caattttacg cagactatct ttctagggtt aaaagcttgg
     3841 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     3901 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     3961 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga
     4021 tccgcatctc aataagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     4081 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4141 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     4201 aggcctaggc ttttgcaaaa agctaacttg tttattgcag cttataatgg ttacaaataa
     4261 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4321 ttgtccaaac tcatcaatgt atcttatcat gtctggatcc gctgcattaa tgaatcggcc
     4381 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4441 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4501 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4561 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4621 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4681 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4741 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4801 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4861 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4921 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4981 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5041 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5101 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5161 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5221 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5281 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5341 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5401 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5461 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5521 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5581 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5641 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5701 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5761 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5821 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5881 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5941 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6001 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6061 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6121 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6181 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6241 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6301 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctg
//
