LOCUS       sequence-313629-        5820 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Actin, Excitation: 488, Emission: 507.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5820)
  AUTHORS   Rizzo MA, Davidson MW, Piston DW
  TITLE     Fluorescent protein tracking and detection: fluorescent protein 
            structure and color variants.
  JOURNAL   Cold Spring Harb Protoc. 2009 Dec;2009(12):pdb.top63. doi: 
            10.1101/pdb.top63.
  PUBMED    20150100
REFERENCE   2  (bases 1 to 5820)
  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: "Cold Spring
            Harb Protoc."; date: "2009-12"; volume: "2009(12):pdb.top63. doi"; 
            pages: " 10.1101/pdb.top63"
COMMENT     Sequence Label: EGFP-Actin-7
FEATURES             Location/Qualifiers
     source          1..5820
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        76..379
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        380..583
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     534..554
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      622..631
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(628..630,631..633,634..1344)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(628..630,631..633,634..1344)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(673..694)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(934..953)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1281..1302
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2623..2744
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2660..2679)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2714..2733
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2751..3206)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2888..2909)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3100..3119
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3233..3337
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3339..3696
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3339..3530
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3362..3382)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3547..3682
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3731..4525
                     /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             3731..4525
                     /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(3785..3804)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4395..4414
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4713..4732)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4757..4804
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5133..5721
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5622..5641
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc
       61 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      121 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      181 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      241 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      301 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      361 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      421 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      481 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      541 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      601 gatccgctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
      661 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
      721 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
      781 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
      841 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
      901 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
      961 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     1021 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     1081 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     1141 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     1201 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     1261 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     1321 ctcggcatgg acgagctgta caagtccgga ctcagatctc gagctatgga tgatgatatc
     1381 gccgcgctcg tcgtcgacaa cggctccggc atgtgcaagg ccggcttcgc gggcgacgat
     1441 gccccccggg ccgtcttccc ctccatcgtg gggcgcccca ggcaccaggg cgtgatggtg
     1501 ggcatgggtc agaaggattc ctatgtgggc gacgaggccc agagcaagag aggcatcctc
     1561 accctgaagt accccatcga gcacggcatc gtcaccaact gggacgacat ggagaaaatc
     1621 tggcaccaca ccttctacaa tgagctgcgt gtggctcccg aggagcaccc cgtgctgctg
     1681 accgaggccc ccctgaaccc caaggccaac cgcgagaaga tgacccagat catgtttgag
     1741 accttcaaca ccccagccat gtacgttgct atccaggctg tgctatccct gtacgcctct
     1801 ggccgtacca ctggcatcgt gatggactcc ggtgacgggg tcacccacac tgtgcccatc
     1861 tacgaggggt atgccctccc ccatgccatc ctgcgtctgg acctggctgg ccgggacctg
     1921 actgactacc tcatgaagat cctcaccgag cgcggctaca gcttcaccac cacggccgag
     1981 cgggaaatcg tgcgtgacat taaggagaag ctgtgctacg tcgccctgga cttcgagcaa
     2041 gagatggcca cggctgcttc cagctcctcc ctggagaaga gctacgagct gcctgacggc
     2101 caggtcatca ccattggcaa tgagcggttc cgctgccctg aggcactctt ccagccttcc
     2161 ttcctgggca tggagtcctg tggcatccac gaaactacct tcaactccat catgaagtgt
     2221 gacgtggaca tccgcaaaga cctgtacgcc aacacagtgc tgtctggcgg caccaccatg
     2281 taccctggca ttgccgacag gatgcagaag gagatcactg ccctggcacc cagcacaatg
     2341 aagatcaaga tcattgctcc tcctgagcgc aagtactccg tgtggatcgg cggctccatc
     2401 ctggcctcgc tgtccacctt ccagcagatg tggatcagca agcaggagta tgacgagtcc
     2461 ggcccctcca tcgtccaccg caaatgcttc tagggatcca ccggatctag ataactgatc
     2521 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     2581 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     2641 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     2701 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag
     2761 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     2821 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     2881 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     2941 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     3001 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     3061 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     3121 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     3181 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     3241 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3301 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     3361 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3421 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3481 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3541 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3601 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     3661 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     3721 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     3781 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     3841 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     3901 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     3961 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4021 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4081 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4141 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4201 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     4261 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4321 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     4381 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     4441 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     4501 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     4561 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4621 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4681 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     4741 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     4801 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     4861 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     4921 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     4981 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5041 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5101 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5161 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5221 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5281 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5341 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5401 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5461 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5521 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5581 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5641 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5701 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5761 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
//
