LOCUS       sequence-54307-e        6167 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 6167)
  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 6167)
  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..6167
                     /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)."
     enhancer        499..802
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        805..1080
                     /label=chicken beta-actin promoter
     intron          1081..2089
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2013..2035
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2097..2116
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2144..2821
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             2144..2821
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(2274..2292)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2542..2561
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2736..2759
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2825..2844)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2890..2945
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2944..2963)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   3613..3647
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(3670..3686)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3670..3686)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3683..3705)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3694..3710
                     /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(3718..3724,3725..3742,3743..3748))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3763..3784
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3843..4038
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3889..4024
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4044..4178
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4081..4100)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4135..4154
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4246..4263)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4417..5005)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4497..4516)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5176..5967,5968..6036))
                     /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     5799..5818
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6037..6141)
                     /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 gttattaata gtaatcaatt acggggtcat tagttcatag
      481 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      541 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      601 gactttccat tgacgtcaat gggtggacta tttacggtaa actgcccact tggcagtaca
      661 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      721 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      781 attagtcatc gctattacca tgggtcgagg tgagccccac gttctgcttc actctcccca
      841 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      901 cgatgggggc gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg
      961 ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
     1021 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
     1081 ggagtcgctg cgttgccttc gccccgtgcc ccgctccgcg ccgcctcgcg ccgcccgccc
     1141 cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg
     1201 ggctgtaatt agcgcttggt ttaatgacgg ctcgtttctt ttctgtggct gcgtgaaagc
     1261 cttaaagggc tccgggaggg ccctttgtgc gggggggagc ggctcggggg gtgcgtgcgt
     1321 gtgtgtgtgc gtggggagcg ccgcgtgcgg cccgcgctgc ccggcggctg tgagcgctgc
     1381 gggcgcggcg cggggctttg tgcgctccgc gtgtgcgcga ggggagcgcg gccgggggcg
     1441 gtgccccgcg gtgcgggggg gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt
     1501 gggggggtga gcagggggtg tgggcgcggc ggtcgggctg taaccccccc ctgcaccccc
     1561 ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtgcg gggcgtggcg
     1621 cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc
     1681 cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg ccccggagcg ccggcggctg
     1741 tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg
     1801 acttcctttg tcccaaatct ggcggagccg aaatctggga ggcgccgccg caccccctct
     1861 agcgggcgcg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg gagggccttc
     1921 gtgcgtcgcc gcgccgccgt ccccttctcc atctccagcc tcggggctgc cgcaggggga
     1981 cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg
     2041 ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc tcctgggcaa
     2101 cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcatggcct cctccgagga
     2161 cgtcatcaag gagttcatgc gcttcaaggt gcgcatggag ggctccgtga acggccacga
     2221 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     2281 gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcca
     2341 gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc
     2401 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     2461 cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg
     2521 caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     2581 caccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaaga tgaggctgaa
     2641 gctgaaggac ggcggccact acgacgccga ggtcaagacc acctacatgg ccaagaagcc
     2701 cgtgcagctg cccggcgcct acaagaccga catcaagctg gacatcacct cccacaacga
     2761 ggactacacc atcgtggaac agtacgagcg cgccgagggc cgccactcca ccggcgccta
     2821 aagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga
     2881 cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc
     2941 tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg
     3001 tttagagttt ggcaacatat gccatatgct ggctgccatg aacaaaggtg gctataaaga
     3061 ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg
     3121 acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta
     3181 aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc
     3241 atagctgtcc ctcttctctt atgaagatcc ctcgacctgc aggctccagc ctacacaatc
     3301 gctcaagacg tgtaatgctt ttattatata ttagtcacga tatctataac aagaaaatat
     3361 atatataata agttatcacg taagtagaac atgaaataac aatataatta tcgtatgagt
     3421 taaatcttaa aagtcacgta aaagataatc atgcgtcatt ttgactcacg cggtcgttat
     3481 agttcaaaat cagtgacact taccgcattg acaagcacgc ctcacgggag ctccaagcgg
     3541 cgactgagat gtcctaaatg cacagcgacg gattcgcgct atttagaaag agagagcaat
     3601 atttcaagaa tgcatgcgtc aattttacgc agactatctt tctagggtta aaagcttggc
     3661 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3721 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     3781 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat
     3841 ccgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3901 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3961 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4021 ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa
     4081 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4141 tgtccaaact catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca
     4201 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4261 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4321 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4381 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4441 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4501 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4561 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     4621 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4681 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4741 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4801 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     4861 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4921 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4981 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5041 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5101 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5161 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5221 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5281 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5341 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5401 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5461 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5521 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5581 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5641 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5701 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5761 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5821 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5881 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5941 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6001 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6061 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6121 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctg
//
