LOCUS       sequence-122868-        6948 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 6948)
  AUTHORS   Murakoshi H, Lee SJ, Yasuda R
  TITLE     Highly sensitive and quantitative FRET-FLIM imaging in single 
            dendritic spines using improved non-radiative YFP.
  JOURNAL   Brain Cell Biol. 2008 Aug . 36(1-4):31-42.
  PUBMED    18512154
REFERENCE   2  (bases 1 to 6948)
  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: "Brain Cell 
            Biol. 2008 Aug . 36(1-4):31-42."
COMMENT     Sequence Label: pCAG-mGFP-Actin
FEATURES             Location/Qualifiers
     source          1..6948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        373..650
                     /label=chicken beta-actin promoter
     intron          651..1671
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1591..1613
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1679..1698
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1742..1751
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1748..1750,1751..1753,1754..2464)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1793..1814)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2054..2073)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2401..2422
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3743..3864
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3780..3799)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3834..3853
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3871..4326)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4008..4029)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4220..4239
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4353..4457
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4459..4816
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4459..4650
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4482..4502)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4667..4802
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4851..5645
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4851..5645
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4905..4924)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5515..5534
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5833..5852)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5877..5924
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6253..6841
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6742..6761
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct
      421 ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg
      481 gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg ggcggggcga
      541 ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg
      601 cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg
      661 cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct
      721 gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta
      781 attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg
      841 ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt
      901 gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg
      961 gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc
     1021 cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg
     1081 ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc
     1141 gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc
     1201 tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc
     1261 gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag
     1321 gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc
     1381 ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg
     1441 ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg
     1501 cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg
     1561 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
     1621 ctagctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
     1681 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattggctag cgctaccggt
     1741 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     1801 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     1861 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     1921 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     1981 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     2041 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     2101 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     2161 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     2221 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     2281 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     2341 caaccactac ctgagcaccc agtccaagct gagcaaagac cccaacgaga agcgcgatca
     2401 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     2461 caagtccgga ctcagatctc gagctatgga tgatgatatc gccgcgctcg tcgtcgacaa
     2521 cggctccggc atgtgcaagg ccggcttcgc gggcgacgat gccccccggg ccgtcttccc
     2581 ctccatcgtg gggcgcccca ggcaccaggg cgtgatggtg ggcatgggtc agaaggattc
     2641 ctatgtgggc gacgaggccc agagcaagag aggcatcctc accctgaagt accccatcga
     2701 gcacggcatc gtcaccaact gggacgacat ggagaaaatc tggcaccaca ccttctacaa
     2761 tgagctgcgt gtggctcccg aggagcaccc cgtgctgctg accgaggccc ccctgaaccc
     2821 caaggccaac cgcgagaaga tgacccagat catgtttgag accttcaaca ccccagccat
     2881 gtacgttgct atccaggctg tgctatccct gtacgcctct ggccgtacca ctggcatcgt
     2941 gatggactcc ggtgacgggg tcacccacac tgtgcccatc tacgaggggt atgccctccc
     3001 ccatgccatc ctgcgtctgg acctggctgg ccgggacctg actgactacc tcatgaagat
     3061 cctcaccgag cgcggctaca gcttcaccac cacggccgag cgggaaatcg tgcgtgacat
     3121 taaggagaag ctgtgctacg tcgccctgga cttcgagcaa gagatggcca cggctgcttc
     3181 cagctcctcc ctggagaaga gctacgagct gcctgacggc caggtcatca ccattggcaa
     3241 tgagcggttc cgctgccctg aggcactctt ccagccttcc ttcctgggca tggagtcctg
     3301 tggcatccac gaaactacct tcaactccat catgaagtgt gacgtggaca tccgcaaaga
     3361 cctgtacgcc aacacagtgc tgtctggcgg caccaccatg taccctggca ttgccgacag
     3421 gatgcagaag gagatcactg ccctggcacc cagcacaatg aagatcaaga tcattgctcc
     3481 tcctgagcgc aagtactccg tgtggatcgg cggctccatc ctggcctcgc tgtccacctt
     3541 ccagcagatg tggatcagca agcaggagta tgacgagtcc ggcccctcca tcgtccaccg
     3601 caaatgcttc tagggatcca ccggatctag ataactgatc ataatcagcc ataccacatt
     3661 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     3721 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     3781 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3841 gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat
     3901 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     3961 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     4021 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     4081 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     4141 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     4201 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     4261 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     4321 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4381 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4441 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     4501 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4561 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4621 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4681 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4741 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4801 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     4861 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4921 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4981 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     5041 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     5101 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     5161 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     5221 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     5281 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     5341 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     5401 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     5461 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     5521 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     5581 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     5641 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     5701 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     5761 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     5821 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     5881 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     5941 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     6001 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     6061 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     6121 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6181 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6241 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6301 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6361 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     6421 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6481 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6541 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6601 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6661 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6721 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6781 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6841 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6901 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgc
//
