LOCUS       sequence-176277-        5275 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses C-terminally HA-tagged Arf4(T31N) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5275)
  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 5275)
  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(T31N)-HA
FEATURES             Location/Qualifiers
     source          1..5275
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      57..66
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             621..647
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(735..754)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    800..855
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(854..873)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1216..1232)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1216..1232)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1229..1251)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1240..1256
                     /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(1264..1270,1271..1288,1289..1294))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1309..1330
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1389..1584
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1435..1570
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1590..1724
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1627..1646)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1681..1700
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1792..1809)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1963..2551)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2043..2062)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2722..3513,3514..3582))
                     /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     3345..3364
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3583..3687)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3999..4802
                     /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(4062..4081)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4672..4691
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4809..4857
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     enhancer        4867..5246
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgctgtctc atcattttgg caaagaattt gctagcgagc tcgcggccgc agatccgcca
       61 ccatgggcct cactatctcc tccctcttct cccgactatt tggcaagaag cagatgcgca
      121 ttttgatggt tggattggat gctgctggca agaacaccat tctgtataaa ctgaagttag
      181 gggagatagt caccaccatt cctaccattg gttttaatgt ggaaacagta gaatataaga
      241 acatttgttt cacagtatgg gatgttggtg gtcaagatag aattaggcct ctctggaagc
      301 attacttcca gaatacccag ggtcttattt ttgtggtaga tagcaacgat cgtgaaagaa
      361 ttcaggaagt agcagatgag ctgcagaaaa tgcttctggt agatgaattg agagatgcag
      421 tgctgctact ttttgcaaac aaacaggatt tgccaaatgc tatggccatc agtgaaatga
      481 cagataaact agggcttcag tctcttcgta acagaacatg gtatgttcaa gccacttgtg
      541 caacacaagg aactggtctg tatgaaggac ttgactggct gtcaaatgag ctttcaaaac
      601 gtctcgaggt accgaattcg tacccatacg acgtcccaga ctacgcttaa ttcactcctc
      661 aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat
      721 accactgaga tcgatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt
      781 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
      841 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat
      901 gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt
      961 tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat
     1021 agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct
     1081 ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga
     1141 ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacct gcagcccaag
     1201 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     1261 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     1321 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     1381 gcggatccgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     1441 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     1501 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     1561 tttggaggcc taggcttttg caaaaagcta acttgtttat tgcagcttat aatggttaca
     1621 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     1681 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatccgctgc attaatgaat
     1741 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     1801 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     1861 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     1921 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1981 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2041 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2101 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2161 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2221 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2281 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2341 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2401 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2461 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2521 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2581 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2641 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2701 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     2761 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     2821 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     2881 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     2941 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3001 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3061 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3121 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3181 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3241 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3301 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     3361 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3421 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3481 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3541 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     3601 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3661 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgggtcgag
     3721 cagtgtggtt ttcaagagga agcaaaaagc ctctccaccc aggcctggaa tgtttccacc
     3781 caatgtcgag cagtgtggtt ttgcaagagg aagcaaaaag cctctccacc caggcctgga
     3841 atgtttccac ccaatgtcga gcaaaccccg cccagcgtct tgtcattggc gaattaattc
     3901 gaacacgcag atgcagtcgg ggcggcgcgg tcccaggtcc acttcgcata ttaaggtgac
     3961 gcgtgtggcc tcgaacaccg agcgaccctg cagccaatat gggatcggcc attgaacaag
     4021 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4081 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4141 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     4201 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4261 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4321 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4381 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4441 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4501 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gatgatctcg
     4561 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4621 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4681 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4741 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4801 gaggggatcg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcgga
     4861 tccgtcgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
     4921 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     4981 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     5041 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     5101 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     5161 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     5221 acgtattagt catcgctatt accatggtcg aggtgagccc cacgttctgc ttcac
//
