LOCUS       sequence-176275-        5965 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses C-terminally mCherry-tagged Arf4(WT) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5965)
  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 5965)
  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)-mCherry
FEATURES             Location/Qualifiers
     source          1..5965
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      8..17
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      572..581
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             578..1255
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             578..1255
                     /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(708..726)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     976..995
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1170..1193
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1343..1362)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1408..1463
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1462..1481)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1824..1840)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1824..1840)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1837..1859)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1848..1864
                     /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(1872..1878,1879..1896,1897..1902))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1917..1938
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1997..2192
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2043..2178
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2198..2332
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2235..2254)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2289..2308
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2400..2417)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2571..3159)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2651..2670)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3330..4121,4122..4190))
                     /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     3953..3972
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4191..4295)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4607..5410
                     /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(4670..4689)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5280..5299
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5417..5465
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     enhancer        5475..5854
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cagatccgcc accatgggcc tcactatctc ctccctcttc tcccgactat ttggcaagaa
       61 gcagatgcgc attttgatgg ttggattgga tgctgctggc aagacaacca ttctgtataa
      121 actgaagtta ggggagatag tcaccaccat tcctaccatt ggttttaatg tggaaacagt
      181 agaatataag aacatttgtt tcacagtatg ggatgttggt ggtcaagata gaattaggcc
      241 tctctggaag cattacttcc agaataccca gggtcttatt tttgtggtag atagcaacga
      301 tcgtgaaaga attcaggaag tagcagatga gctgcagaaa atgcttctgg tagatgaatt
      361 gagagatgca gtgctgctac tttttgcaaa caaacaggat ttgccaaatg ctatggccat
      421 cagtgaaatg acagataaac tagggcttca gtctcttcgt aacagaacat ggtatgttca
      481 agccacttgt gcaacacaag gaactggtct gtatgaagga cttgactggc tgtcaaatga
      541 gctttcaaaa cgtctcgagg gacttaggtc tgccaccatg gcctcctccg aggacgtcat
      601 caaggagttc atgcgcttca aggtgcgcat ggagggctcc gtgaacggcc acgagttcga
      661 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
      721 gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tccagtacgg
      781 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc
      841 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
      901 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
      961 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccaccga
     1021 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagatgaggc tgaagctgaa
     1081 ggacggcggc cactacgacg ccgaggtcaa gaccacctac atggccaaga agcccgtgca
     1141 gctgcccggc gcctacaaga ccgacatcaa gctggacatc acctcccaca acgaggacta
     1201 caccatcgtg gaacagtacg agcgcgccga gggccgccac tccaccggcg cctaacaatt
     1261 cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc
     1321 tcacaaatac cactgagatc gatctttttc cctctgccaa aaattatggg gacatcatga
     1381 agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt
     1441 gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac
     1501 atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga
     1561 acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct
     1621 tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat
     1681 ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc
     1741 tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacctgc
     1801 agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     1861 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     1921 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     1981 tcgtgccagc ggatccgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     2041 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2101 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     2161 gaggcttttt tggaggccta ggcttttgca aaaagctaac ttgtttattg cagcttataa
     2221 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     2281 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tccgctgcat
     2341 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     2401 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     2461 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     2521 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2581 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2641 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2701 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2761 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2821 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2881 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2941 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3001 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3061 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3121 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3181 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3241 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3301 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3361 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3421 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3481 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3541 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3601 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     3661 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3721 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3781 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3841 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3901 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3961 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4021 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4081 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4141 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     4201 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4261 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     4321 gggtcgagca gtgtggtttt caagaggaag caaaaagcct ctccacccag gcctggaatg
     4381 tttccaccca atgtcgagca gtgtggtttt gcaagaggaa gcaaaaagcc tctccaccca
     4441 ggcctggaat gtttccaccc aatgtcgagc aaaccccgcc cagcgtcttg tcattggcga
     4501 attaattcga acacgcagat gcagtcgggg cggcgcggtc ccaggtccac ttcgcatatt
     4561 aaggtgacgc gtgtggcctc gaacaccgag cgaccctgca gccaatatgg gatcggccat
     4621 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4681 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4741 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     4801 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     4861 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     4921 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4981 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     5041 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     5101 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     5161 tgatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     5221 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     5281 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     5341 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     5401 gttcttctga ggggatcggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt
     5461 tgttcggatc cgtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
     5521 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     5581 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     5641 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     5701 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     5761 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     5821 gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt
     5881 cactctcccc atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt
     5941 attttgtgca gcgatggggg cgggg
//
