LOCUS       sequence-177248-        5997 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 5997)
  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 5997)
  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: PBMBP-eGFP
FEATURES             Location/Qualifiers
     source          1..5997
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   11..73
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    370..403
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      1839..1848
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1845..1847,1848..1850,1851..2564)
                     /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(1845..1847,1848..1850,1851..2561)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1890..1911)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2151..2170)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2498..2519
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2651..2670)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2716..2771
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2770..2789)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   3442..3476
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(3499..3515)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3499..3515)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3512..3534)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3523..3539
                     /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(3547..3553,3554..3571,3572..3577))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3592..3613
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3672..3867
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3718..3853
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3873..4007
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3910..3929)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3964..3983
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4075..4092)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4246..4834)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4326..4345)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5005..5796,5797..5865))
                     /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     5628..5647
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5866..5970)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacttaa ccctagaaag ataatcatat tgtgacgtac gttaaagata atcatgcgta
       61 aaattgacgc atgtgtttta tcgatctgta tatcgaggtt tatttattaa tttgaataga
      121 tattaagttt tattatattt acacttacat actaataata aattcaacaa acaatttatt
      181 tatgtttatt tatttattaa aaaaaaacaa aaactcaaaa tttcttctat aaagtaacaa
      241 aacttttaaa cattctctct tttacaaaaa taaacttatt ttgtacttta aaaacagtca
      301 tgttgtatta taaaataagt aattagctta acttatacat aatagaaaca aattatactt
      361 attaatcgca taacttcgta taatgtatgc tatacgaagt tatgcgatta agggatctgt
      421 agggcgcagt agtccaacta gtaagtttcg tgagaaaagg gaccagatct tattcctcac
      481 cgtggcttta acacttagag aaaatgcatc ccctctaatc aataagtcat cgacagtggg
      541 tagatggagg aacggcagtg cgtagtagga tgcgtgcaag catagtctcg tgcatgggtg
      601 catagatcgc tgggcaggtg gacaaggtgg gggtggataa agaagtgggt agatgattga
      661 tgttaggtaa atatcactgg gtggacagat gggtggtagg tggatggatg gttagaatag
      721 tcagaagagg gatggattga taaggtgaac agatgataaa tgggtgatag actggaaggg
      781 ttgtcaaaag aggataaggg aagtgtgagc tagccgtatt tctaaggtca gtaatagagt
      841 tgggagaaga ggttaagtta catccattta aacctcacac gaagctgaga gggaatggac
      901 ttgctgccgt tggtgaggaa agcgttgcat ttcccgtgtg cttggttgtg aagtgctcag
      961 gtcccacatg aagcagtcag gttactgcgg cttacagagg agccagatcc aaatgccccg
     1021 agtaagcacg tccccgagcc agaggcctcc agcggaatcc gggagaggga ttgctcagtg
     1081 ccctgcttcc ctggactgta agctgcagaa agatgtggga agtcctgttc tccactgaga
     1141 acactaaaag caccttttgt caaacgaccg cttcacatct ggggcttgtg cactggtggc
     1201 cttttaaacc agagacaacc cacaagatac ctaacctgcg gggctctctg gtacagtgag
     1261 caactcagga aatgctttgg cttgattgct gtgggctctc aggccatcgc cctctggagt
     1321 ggttctttta atgagaacct gaagattggc ccctgagcca tgtataccaa gcaagctcaa
     1381 tccaggttag ctccctctgg ttggggcaag ctaacgtgct ccttgggccc cgcgcgtaac
     1441 tgtgcgtttt ataggagaca gctagttcaa gaccccagga agaaagcggc tttgtccccc
     1501 tctaggcctc gtacaggccc acattcatat ctcattgttg ttgcagggga ggcagatgcg
     1561 atccagaaca atgggacctc ggctgaggac acggcggtga cagactccaa gcacacagca
     1621 gacccaaaga ataactggca aggcgcccac ccagctgacc cagggaaccg cccccacttg
     1681 atccgcctct tttcccgaga tgccccggga agggaggaca acaccttcaa agacaggccc
     1741 tcagagtccg acgagcttca gaccatccaa gaagatccca cagcagcttc cgaagaattc
     1801 tgcagtcgac ggtaccgcgg gcccgggatc caccggtcgc caccatggtg agcaagggcg
     1861 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     1921 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     1981 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     2041 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     2101 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     2161 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     2221 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     2281 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     2341 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     2401 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     2461 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     2521 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc cgcactcctc
     2581 aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat
     2641 accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc
     2701 atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt
     2761 tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt
     2821 atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc
     2881 tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa
     2941 aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa
     3001 catccctaaa attttcctta catgttttac tagccagatt tttcctcctc tcctgactac
     3061 tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcag agctccagcc
     3121 tacacaatcg ctcaagacgt gtaatgcttt tattatatat tagtcacgat atctataaca
     3181 agaaaatata tatataataa gttatcacgt aagtagaaca tgaaataaca atataattat
     3241 cgtatgagtt aaatcttaaa agtcacgtaa aagataatca tgcgtcattt tgactcacgc
     3301 ggtcgttata gttcaaaatc agtgacactt accgcattga caagcacgcc tcacgggagc
     3361 tccaagcggc gactgagatg tcctaaatgc acagcgacgg attcgcgcta tttagaaaga
     3421 gagagcaata tttcaagaat gcatgcgtca attttacgca gactatcttt ctagggttaa
     3481 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     3541 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     3601 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     3661 ccagcggatc cgcatctcaa taagtcagca accatagtcc cgcccctaac tccgcccatc
     3721 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3781 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3841 ttttttggag gcctaggctt ttgcaaaaag ctaacttgtt tattgcagct tataatggtt
     3901 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3961 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatccgc tgcattaatg
     4021 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4081 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4141 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4201 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4261 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4321 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4381 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4441 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4501 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4561 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4621 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4681 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4741 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4801 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4861 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4921 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4981 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5041 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5101 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5161 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5221 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5281 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5341 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5401 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5461 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5521 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5581 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5641 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5701 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5761 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5821 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5881 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     5941 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgg
//
