LOCUS       sequence-176233-        5952 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally HA-tagged Arf6(WT) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5952)
  AUTHORS   Makyio H, Ohgi M, Takei T, Takahashi S, Takatsu H, Katoh Y, Hanai A,
            Ueda T, Kanaho Y, Xie Y, Shin HW, Kamikubo H, Kataoka M, Kawasaki M,
            Kato R, Wakatsuki S, Nakayama K
  TITLE     Structural basis for Arf6-MKLP1 complex formation on the Flemming 
            body responsible for cytokinesis.
  JOURNAL   EMBO J. 2012 May 30;31(11):2590-603. doi: 10.1038/emboj.2012.89. 
            Epub 2012 Apr 20.
  PUBMED    22522702
REFERENCE   2  (bases 1 to 5952)
  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: 
            "10.1038/emboj.2012.89"; journalName: "EMBO J"; date: "2012-05-30- 
            30"; volume: "31"; issue: "11"; pages: "2590-603"
COMMENT     Sequence Label: pcDNA3/hArf6(WT)-HA
FEATURES             Location/Qualifiers
     source          1..5952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      92..520
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(179..198)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     389..410
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(529..549)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        534..863
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      714..849
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     776..795
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             930..1724
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(984..1003)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1594..1613
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1898..2019
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1935..1954)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1989..2008
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2068..2084)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2068..2084)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2081..2103)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2092..2108
                     /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(2116..2122,2123..2140,2141..2146))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2161..2182
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2299..2316)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2470..3058)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2550..2569)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3229..4020,4021..4089))
                     /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     3852..3871
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4090..4194)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4269..4288)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4460..4839
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4840..5043
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4994..5014
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5088..5107
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5088..5106
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      5138..5147
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5675..5701
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(5704..5720)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5704..5720)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(5730..5748)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5731..5748)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5768..5785)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    join(5774..5952,1..46)
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
       61 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
      121 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
      181 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
      241 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
      301 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
      361 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
      421 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
      481 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
      541 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
      601 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
      661 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
      721 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
      781 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
      841 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
      901 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
      961 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     1021 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     1081 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     1141 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     1201 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     1261 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     1321 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     1381 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     1441 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     1501 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     1561 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     1621 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     1681 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     1741 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     1801 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     1861 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     1921 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     1981 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     2041 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     2101 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     2161 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     2221 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2281 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     2341 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2401 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2461 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2521 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2581 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2641 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2701 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2761 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2821 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2881 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2941 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3001 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3061 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3121 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3181 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3241 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3301 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3361 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3421 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     3481 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     3541 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     3601 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     3661 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     3721 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     3781 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     3841 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     3901 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     3961 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4021 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4081 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4141 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4201 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     4261 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     4321 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     4381 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
     4441 cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg
     4501 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     4561 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     4621 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     4681 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     4741 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     4801 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     4861 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     4921 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     4981 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     5041 gctctctggc taactagaga acccactgct tactggctta tcgaaattaa tacgactcac
     5101 tatagggaga cccaagcttg gtaccgagct cggatctgcc accatgggga aggtgctatc
     5161 caaaatcttc gggaacaagg aaatgcggat cctcatgttg ggcctggacg cggccggcaa
     5221 gacaacaatc ctgtacaagt tgaagctggg ccagtcggtg accaccattc ccactgtggg
     5281 tttcaacgtg gagacggtga cttacaaaaa tgtcaagttc aacgtatggg atgtgggcgg
     5341 ccaggacaag atccggccgc tctggcggca ttactacact gggacccaag gtctcatctt
     5401 cgtagtggac tgcgccgacc gcgaccgcat cgatgaggct cgccaggagc tgcaccgcat
     5461 tatcaatgac cgggagatga gggacgccat aatcctcatc ttcgccaaca agcaggacct
     5521 gcccgatgcc atgaaacccc acgagatcca ggagaaactg ggcctgaccc ggattcggga
     5581 caggaactgg tatgtgcagc cctcctgtgc cacctcaggg gacggactct atgaggggct
     5641 cacatggtta acctctaact acaaatctct cgagtatcca tacgatgtac cagactacgc
     5701 agagatccac tagttctaga gggccctatt ctatagtgtc acctaaatgc tagagctcgc
     5761 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     5821 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     5881 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     5941 aagggggagg at
//
