LOCUS       sequence-176276-        5334 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses C-terminally HA-tagged Arf4(Q71L) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5334)
  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 5334)
  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(Q71L)-HA
FEATURES             Location/Qualifiers
     source          1..5334
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      39..48
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             603..629
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(717..736)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    782..837
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(836..855)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1198..1214)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1198..1214)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1211..1233)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1222..1238
                     /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(1246..1252,1253..1270,1271..1276))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1291..1312
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1371..1566
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1417..1552
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1572..1706
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1609..1628)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1663..1682
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1774..1791)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1945..2533)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2025..2044)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2704..3495,3496..3564))
                     /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     3327..3346
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3565..3669)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3981..4784
                     /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(4044..4063)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4654..4673
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4791..4839
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     enhancer        4849..5228
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggcaaagaat ttgctagcga gctcgcggcc gcagatccgc caccatgggc ctcactatct
       61 cctccctctt ctcccgacta tttggcaaga agcagatgcg cattttgatg gttggattgg
      121 atgctgctgg caagacaacc attctgtata aactgaagtt aggggagata gtcaccacca
      181 ttcctaccat tggttttaat gtggaaacag tagaatataa gaacatttgt ttcacagtat
      241 gggatgttgg tggtctggat agaattaggc ctctctggaa gcattacttc cagaataccc
      301 agggtcttat ttttgtggta gatagcaacg atcgtgaaag aattcaggaa gtagcagatg
      361 agctgcagaa aatgcttctg gtagatgaat tgagagatgc agtgctgcta ctttttgcaa
      421 acaaacagga tttgccaaat gctatggcca tcagtgaaat gacagataaa ctagggcttc
      481 agtctcttcg taacagaaca tggtatgttc aagccacttg tgcaacacaa ggaactggtc
      541 tgtatgaagg acttgactgg ctgtcaaatg agctttcaaa acgtctcgag gtaccgaatt
      601 cgtacccata cgacgtccca gactacgctt aattcactcc tcaggtgcag gctgcctatc
      661 agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga gatcgatctt
      721 tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc
      781 taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc
      841 ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt
      901 ttggcaacat atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat
      961 cagtatatga aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga
     1021 ggttagattt tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt
     1081 tccttacatg ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct
     1141 gtccctcttc tcttatggag atccctcgac ctgcagccca agcttggcgt aatcatggtc
     1201 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     1261 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     1321 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat
     1381 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     1441 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     1501 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     1561 tgcaaaaagc taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     1621 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     1681 tcaatgtatc ttatcatgtc tggatccgct gcattaatga atcggccaac gcgcggggag
     1741 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     1801 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     1861 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     1921 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1981 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2041 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2101 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2161 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2221 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2281 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2341 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     2401 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     2461 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     2521 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     2581 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     2641 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     2701 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     2761 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     2821 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     2881 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     2941 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     3001 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     3061 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     3121 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     3181 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     3241 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     3301 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     3361 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     3421 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     3481 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     3541 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     3601 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     3661 ggttccgcgc acatttcccc gaaaagtgcc acctgggtcg agcagtgtgg ttttcaagag
     3721 gaagcaaaaa gcctctccac ccaggcctgg aatgtttcca cccaatgtcg agcagtgtgg
     3781 ttttgcaaga ggaagcaaaa agcctctcca cccaggcctg gaatgtttcc acccaatgtc
     3841 gagcaaaccc cgcccagcgt cttgtcattg gcgaattaat tcgaacacgc agatgcagtc
     3901 ggggcggcgc ggtcccaggt ccacttcgca tattaaggtg acgcgtgtgg cctcgaacac
     3961 cgagcgaccc tgcagccaat atgggatcgg ccattgaaca agatggattg cacgcaggtt
     4021 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4081 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4141 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     4201 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4261 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4321 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4381 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4441 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4501 tcgccaggct caaggcgcgc atgcccgacg gcgatgatct cgtcgtgacc catggcgatg
     4561 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4621 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4681 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4741 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgaggggat cggcaataaa
     4801 aagacagaat aaaacgcacg ggtgttgggt cgtttgttcg gatccgtcga cattgattat
     4861 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     4921 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     4981 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     5041 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     5101 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     5161 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     5221 ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc
     5281 cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg gggg
//
