LOCUS       sequence-176237-        5251 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally EGFP-tagged Arf5(WT) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5251)
  AUTHORS   Makyio H, Ohgi M, Takei T, Takahashi S, Takatsu H, Katoh Y, Hanai A,
            Ueda T, Kanaho Y, Xie Y, Shin HW, Kamikubo H, Kataoka M, Kawasaki M,
            Kato R, Wakatsuki S, Nakayama K
  TITLE     Structural basis for Arf6-MKLP1 complex formation on the Flemming 
            body responsible for cytokinesis.
  JOURNAL   EMBO J. 2012 May 30;31(11):2590-603. doi: 10.1038/emboj.2012.89. 
            Epub 2012 Apr 20.
  PUBMED    22522702
REFERENCE   2  (bases 1 to 5251)
  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"; doi: 
            "10.1038/emboj.2012.89"; journalName: "EMBO J"; date: "2012-05-30- 
            30"; volume: "31"; issue: "11"; pages: "2590-603"
COMMENT     Sequence Label: pEGFP-N3/hArf5(WT)-EGFP
FEATURES             Location/Qualifiers
     source          1..5251
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             85..879
                     /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             85..879
                     /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(139..158)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     749..768
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1067..1086)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1111..1158
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1487..2075
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1976..1995
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2250..2553
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2554..2757
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2708..2728
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2804..2813
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3380..3389
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3386..3388,3389..3391,3392..4105)
                     /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(3386..3388,3389..3391,3392..4102)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3431..3452)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3692..3711)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4039..4060
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4228..4349
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4265..4284)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4319..4338
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4356..4811)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4493..4514)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4705..4724
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4838..4942
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(4944..5251,1..50)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4944..5135
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4967..4987)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(5152..5251,1..36)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
       61 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
      121 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
      181 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
      241 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
      301 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
      361 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
      421 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
      481 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
      541 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
      601 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
      661 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
      721 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
      781 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
      841 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
      901 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
      961 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     1021 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     1081 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     1141 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     1201 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     1261 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     1321 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     1381 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     1441 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     1501 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     1561 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     1621 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     1681 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     1741 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     1801 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     1861 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     1921 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     1981 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     2041 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     2101 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     2161 ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca
     2221 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     2281 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     2341 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     2401 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     2461 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     2521 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
     2581 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     2641 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     2701 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt
     2761 ttagtgaacc gtcagatccg ctagcgctac cggactcaga tccgccacca tgggcctcac
     2821 cgtgtccgcg ctcttttcgc ggatcttcgg gaagaagcag atgcggattc tcatggttgg
     2881 cttggatgcg gctggcaaga ccacaatcct gtacaaactg aagttggggg agattgtcac
     2941 caccatccca accataggct tcaatgtaga aacagtggaa tataagaaca tctgtttcac
     3001 agtctgggac gtgggaggcc aggacaagat tcggcctctg tggcggcact acttccagaa
     3061 cactcagggc ctcatctttg tggtggacag taatgaccgg gagcgggtcc aagaatctgc
     3121 tgatgaactc cagaagatgc tgcaggagga cgagctgcgg gatgcagtgc tgctggtatt
     3181 tgccaacaag caggacatgc ccaacgccat gcccgtgagc gagctgactg acaagctggg
     3241 gctacagcac ttacgcagcc gcacgtggta tgtccaggcc acctgtgcca cccaaggcac
     3301 aggtctgtac gatggtctgg actggctgtc ccacgagctg tcaaagcgcc tcgacggtac
     3361 cgcgggcccg ggatccatcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt
     3421 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     3481 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     3541 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     3601 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     3661 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     3721 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     3781 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     3841 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     3901 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     3961 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc
     4021 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     4081 cggcatggac gagctgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc
     4141 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     4201 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     4261 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4321 ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt
     4381 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     4441 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     4501 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     4561 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     4621 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     4681 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     4741 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     4801 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4861 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4921 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     4981 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     5041 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5101 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     5161 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     5221 gcagaggccg aggccgcctc ggcctctgag c
//
