LOCUS       sequence-176279-        6026 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses C-terminally EGFP-tagged Arf4(WT) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6026)
  AUTHORS   Nakai W, Kondo Y, Saitoh A, Naito T, Nakayama K, Shin HW
  TITLE     ARF1 and ARF4 regulate recycling endosomal morphology and retrograde
            transport from endosomes to the Golgi apparatus.
  JOURNAL   Mol Biol Cell. 2013 Aug;24(16):2570-81. doi: 
            10.1091/mbc.E13-04-0197. Epub 2013 Jun 19.
  PUBMED    23783033
REFERENCE   2  (bases 1 to 6026)
  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: 
            "10.1091/mbc.E13-04-0197"; journalName: "Mol Biol Cell"; date: 
            "2013-08"; volume: "24"; issue: "16"; pages: "2570-81"
COMMENT     Sequence Label: pCAG/hArf4(WT)-EGFP
FEATURES             Location/Qualifiers
     source          1..6026
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      20..29
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      596..605
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(602..604,605..607,608..1321)
                     /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(602..604,605..607,608..1318)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(647..668)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(908..927)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1255..1276
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1409..1428)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1474..1529
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1528..1547)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1890..1906)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1890..1906)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1903..1925)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1914..1930
                     /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(1938..1944,1945..1962,1963..1968))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1983..2004
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2063..2258
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2109..2244
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2264..2398
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2301..2320)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2355..2374
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2466..2483)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2637..3225)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2717..2736)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3396..4187,4188..4256))
                     /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     4019..4038
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4257..4361)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4673..5476
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4736..4755)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5346..5365
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5483..5531
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     enhancer        5541..5920
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gctagtttga ctcagatccg ccaccatggg cctcactatc tcctccctct tctcccgact
       61 atttggcaag aagcagatgc gcattttgat ggttggattg gatgctgctg gcaagacaac
      121 cattctgtat aaactgaagt taggggagat agtcaccacc attcctacca ttggttttaa
      181 tgtggaaaca gtagaatata agaacatttg tttcacagta tgggatgttg gtggtcaaga
      241 tagaattagg cctctctgga agcattactt ccagaatacc cagggtctta tttttgtggt
      301 agatagcaac gatcgtgaaa gaattcagga agtagcagat gagctgcaga aaatgcttct
      361 ggtagatgaa ttgagagatg cagtgctgct actttttgca aacaaacagg atttgccaaa
      421 tgctatggcc atcagtgaaa tgacagataa actagggctt cagtctcttc gtaacagaac
      481 atggtatgtt caagccactt gtgcaacaca aggaactggt ctgtatgaag gacttgactg
      541 gctgtcaaat gagctttcaa aacgtctcga gggtaccgcg ggcccgggat ccatcgccac
      601 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
      661 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
      721 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
      781 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
      841 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
      901 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
      961 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     1021 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     1081 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     1141 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1201 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     1261 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     1321 acaattcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc
     1381 cctggctcac aaataccact gagatcgatc tttttccctc tgccaaaaat tatggggaca
     1441 tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa
     1501 tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt
     1561 taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg
     1621 ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc
     1681 attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg
     1741 tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt
     1801 tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg
     1861 acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     1921 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     1981 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     2041 aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc ccgcccctaa
     2101 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     2161 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     2221 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc
     2281 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     2341 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccg
     2401 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     2461 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2521 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2581 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2641 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2701 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2761 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2821 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2881 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2941 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3001 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3061 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3121 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3181 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3241 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3301 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3361 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     3421 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3481 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3541 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3601 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3661 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     3721 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     3781 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     3841 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     3901 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     3961 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4021 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4081 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4141 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4201 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4261 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4321 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4381 ccacctgggt cgagcagtgt ggttttcaag aggaagcaaa aagcctctcc acccaggcct
     4441 ggaatgtttc cacccaatgt cgagcagtgt ggttttgcaa gaggaagcaa aaagcctctc
     4501 cacccaggcc tggaatgttt ccacccaatg tcgagcaaac cccgcccagc gtcttgtcat
     4561 tggcgaatta attcgaacac gcagatgcag tcggggcggc gcggtcccag gtccacttcg
     4621 catattaagg tgacgcgtgt ggcctcgaac accgagcgac cctgcagcca atatgggatc
     4681 ggccattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4741 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4801 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4861 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4921 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4981 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     5041 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     5101 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     5161 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     5221 cggcgatgat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     5281 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     5341 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5401 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     5461 tgacgagttc ttctgagggg atcggcaata aaaagacaga ataaaacgca cgggtgttgg
     5521 gtcgtttgtt cggatccgtc gacattgatt attgactagt tattaatagt aatcaattac
     5581 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     5641 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     5701 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     5761 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     5821 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     5881 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt
     5941 ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt
     6001 ttaattattt tgtgcagcga tggggg
//
