LOCUS       sequence-177244-        5810 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 5810)
  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 5810)
  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: PBGFAP-eGFP
FEATURES             Location/Qualifiers
     source          1..5810
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..160
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(161..229,230..1021)
                     /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     complement(379..398)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1192..1780
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1681..1700
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1934..1951
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    2019..2153
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2043..2062)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2097..2116
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(2159..2354)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2173..2308
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    2413..2434
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2449..2454,2455..2472,2473..2479)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2487..2503
                     /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)."
     primer_bind     2492..2514
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2511..2527
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2511..2527
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   2550..2584
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     3237..3256
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    3255..3310
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     3356..3375
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(join(3462..4175,4176..4178,4179..4181))
                     /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             complement(join(3465..4175,4176..4178,4179..4181))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3507..3528)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3856..3875
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4115..4136
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4178..4187
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(5436..5469)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     repeat_region   join(5766..5810,1..18)
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 atgattatct ttctagggtt aagtcgaccc aggtggcact tttcggggaa atgtgcgcgg
       61 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
      121 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
      181 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
      241 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
      301 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
      361 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
      421 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
      481 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
      541 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
      601 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
      661 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
      721 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
      781 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
      841 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
      901 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
      961 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     1021 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     1081 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     1141 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     1201 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     1261 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     1321 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     1381 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     1441 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     1501 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     1561 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     1621 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     1681 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     1741 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     1801 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     1861 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     1921 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     1981 gcctctcccc gcgcgttggc cgattcatta atgcagcgga tccagacatg ataagataca
     2041 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     2101 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttagctttt
     2161 tgcaaaagcc taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg
     2221 aggcggcctc ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg
     2281 cggaactggg cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg
     2341 acttattgag atgcggatcc gctggcacga caggtttccc gactggaaag cgggcagtga
     2401 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     2461 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     2521 ctatgaccat gattacgcca agcttttaac cctagaaaga tagtctgcgt aaaattgacg
     2581 catgcattct tgaaatattg ctctctcttt ctaaatagcg cgaatccgtc gctgtgcatt
     2641 taggacatct cagtcgccgc ttggagctcc cgtgaggcgt gcttgtcaat gcggtaagtg
     2701 tcactgattt tgaactataa cgaccgcgtg agtcaaaatg acgcatgatt atcttttacg
     2761 tgacttttaa gatttaactc atacgataat tatattgtta tttcatgttc tacttacgtg
     2821 ataacttatt atatatatat tttcttgtta tagatatcgt gactaatata taataaaagc
     2881 attacacgtc ttgagcgatt gtgtaggctg gagctctgca ggtcgaggga tctccataag
     2941 agaagaggga cagctatgac tgggagtagt caggagagga ggaaaaatct ggctagtaaa
     3001 acatgtaagg aaaattttag ggatgttaaa gaaaaaaata acacaaaaca aaatataaaa
     3061 aaaatctaac ctcaagtcaa ggcttttcta tggaataagg aatggacagc agggggctgt
     3121 ttcatatact gatgacctct ttatagccaa cctttgttca tggcagccag catatgggca
     3181 tatgttgcca aactctaaac caaatactca ttctgatgtt ttaaatgatt tgccctccca
     3241 tatgtccttc cgagtgagag acacaaaaaa ttccaacaca ctattgcaat gaaaataaat
     3301 ttcctttatt agccagaagt cagatgctca aggggcttca tgatgtcccc ataatttttg
     3361 gcagagggaa aaagatctca gtggtatttg tgagccaggg cattggccac accagccacc
     3421 accttctgat aggcagcctg cacctgagga gtgcggccgc tttacttgta cagctcgtcc
     3481 atgccgagag tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc
     3541 tcgttggggt ctttgctcag ggcggactgg gtgctcaggt agtggttgtc gggcagcagc
     3601 acggggccgt cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgctgccg
     3661 tcctcgatgt tgtggcggat cttgaagttc accttgatgc cgttcttctg cttgtcggcc
     3721 atgatataga cgttgtggct gttgtagttg tactccagct tgtgccccag gatgttgccg
     3781 tcctccttga agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac
     3841 ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg
     3901 acgtagcctt cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag
     3961 cggctgaagc actgcacgcc gtaggtcagg gtggtcacga gggtgggcca gggcacgggc
     4021 agcttgccgg tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg
     4081 ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg
     4141 ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca tggtggcgac cggtggatcc
     4201 cgggcccgcg gtaccgtcga ctgcagaatt ctgaccaccg tctcggaggc aaaggagcgg
     4261 cgcgcagagg tgatgcgtct ccgctccaac ctgccctgcc tctgctggct cctgggatgt
     4321 cagggccttt atggaggaac gggttggact aaatctgccc ctgacttcct ggaagcaccc
     4381 ccattgaata gcctgagggg gtggagctgg gccccagccc agctaggggg agaactcctc
     4441 tcaccccact gaggtcactg tacccagagt caaggctttc ttgggaacac cagcctggct
     4501 tcgccattga tcaaggaacc ccctttctgg taaatgggtc aagagagggc cgagggaaca
     4561 gcttagtcca taaattgctt gcctgtcaag catgaagacc cacccaagtt ctagctgcag
     4621 aatgaataca aaaggcctgg gttgacagtg catacttgta aactccagtg ctgggcagtg
     4681 aaaatcagca gacttcctgg ctctcactaa ttggcgagtt ccagatccag agagaaatgc
     4741 tctctcagaa aaccaaggca acagtgcctg aggttgacct ctggtctcaa cacatgtatg
     4801 tgcacacaca tgaacatttg tgtgataagt gtgcacgagc gcacacacag agacagagac
     4861 agagagtgca cagctcggtg gggtaatcag cttccagttc ttagagcctg taagccagga
     4921 ccaccccaac tctgtccttc tctgggggcc tgtgagatgc taggaacttg aagaacctat
     4981 tcccttcctc cgggatcaac ctatattaca gtttcctgtg acccccattc tcaacagagt
     5041 cctctcccct gaatctctcc aactctatct tctcattcct ggagttcagg agtggagggc
     5101 taaggttgga gccctctctg tgactcctgg gtcaacttga gcctctggct gtgtgtttag
     5161 agtgcacggc agaaggtatg cacgcagatc ctaatagggt aagacagcag gcttcattcc
     5221 tgagcccctc tgtggcctct tatccagcct agtgggccat ttctccctga tccagcttct
     5281 cgttgccccc caccccccac cccgagcctg gcccctggca gcccctgaca gagtagtttt
     5341 gaatgagttg gctgtgcatg ctggagtctt tctttctgta acaccccgtc ttcctgacta
     5401 gttggactac tgcgccctac agatccctta atcgcataac ttcgtatagc atacattata
     5461 cgaagttatg cgattaataa gtataatttg tttctattat gtataagtta agctaattac
     5521 ttattttata atacaacatg actgttttta aagtacaaaa taagtttatt tttgtaaaag
     5581 agagaatgtt taaaagtttt gttactttat agaagaaatt ttgagttttt gttttttttt
     5641 aataaataaa taaacataaa taaattgttt gttgaattta ttattagtat gtaagtgtaa
     5701 atataataaa acttaatatc tattcaaatt aataaataaa cctcgatata cagatcgata
     5761 aaacacatgc gtcaatttta cgcatgatta tctttaacgt acgtcacaat
//
