LOCUS       sequence-176252-        4212 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses C-terminally HA-tagged Arf4(WT) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4212)
  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 4212)
  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)-HA
FEATURES             Location/Qualifiers
     source          1..4212
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     320..342
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     404..423
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      477..486
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1041..1067
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1155..1174)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1220..1275
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1274..1293)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1636..1652)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1636..1652)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1649..1671)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1660..1676
                     /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(1684..1690,1691..1708,1709..1714))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1729..1750
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1809..2004
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1855..1990
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2010..2144
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2047..2066)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2101..2120
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2212..2229)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2383..2971)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2463..2482)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3142..3933,3934..4002))
                     /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     3765..3784
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4003..4107)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggctcggggg aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc
       61 cgcagccatt gccttttatg gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa
      121 atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg
      181 aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc
      241 cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg
      301 gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagagcctct
      361 gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt
      421 gtgctgtctc atcattttgg caaagaattt gctagcgagc tcgcggccgc agatccgcca
      481 ccatgggcct cactatctcc tccctcttct cccgactatt tggcaagaag cagatgcgca
      541 ttttgatggt tggattggat gctgctggca agacaaccat tctgtataaa ctgaagttag
      601 gggagatagt caccaccatt cctaccattg gttttaatgt ggaaacagta gaatataaga
      661 acatttgttt cacagtatgg gatgttggtg gtcaagatag aattaggcct ctctggaagc
      721 attacttcca gaatacccag ggtcttattt ttgtggtaga tagcaacgat cgtgaaagaa
      781 ttcaggaagt agcagatgag ctgcagaaaa tgcttctggt agatgaattg agagatgcag
      841 tgctgctact ttttgcaaac aaacaggatt tgccaaatgc tatggccatc agtgaaatga
      901 cagataaact agggcttcag tctcttcgta acagaacatg gtatgttcaa gccacttgtg
      961 caacacaagg aactggtctg tatgaaggac ttgactggct gtcaaatgag ctttcaaaac
     1021 gtctcgaggt accgaattcg tacccatacg acgtcccaga ctacgcttaa ttcactcctc
     1081 aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat
     1141 accactgaga tcgatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt
     1201 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     1261 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat
     1321 gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt
     1381 tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat
     1441 agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct
     1501 ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga
     1561 ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacct gcagcccaag
     1621 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     1681 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     1741 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     1801 gcggatccgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     1861 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     1921 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     1981 tttggaggcc taggcttttg caaaaagcta acttgtttat tgcagcttat aatggttaca
     2041 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     2101 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatccgctgc attaatgaat
     2161 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     2221 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2281 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2341 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2401 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2461 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2521 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2581 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2641 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2701 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2761 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2821 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2881 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2941 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3001 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3061 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3121 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3181 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     3241 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     3301 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3361 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3421 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3481 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3541 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3601 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3661 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3721 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     3781 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3841 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3901 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3961 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4021 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4081 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgggtcgag
     4141 cagtgtggtt ttcaagagga agcaaaaagc ctctccaccc aggcctggaa tgtttccacc
     4201 caatgtcgag ca
//
