LOCUS       sequence-176235-        5952 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally HA-tagged Arf6(Q67L) 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(Q67L)-HA
FEATURES             Location/Qualifiers
     source          1..5952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     33..54
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(173..193)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        178..507
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      358..493
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     420..439
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             574..1368
                     /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(628..647)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1238..1257
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1542..1663
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1579..1598)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1633..1652
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1712..1728)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1712..1728)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1725..1747)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1736..1752
                     /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(1760..1766,1767..1784,1785..1790))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1805..1826
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1943..1960)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2114..2702)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2194..2213)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2873..3664,3665..3733))
                     /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     3496..3515
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3734..3838)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3913..3932)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4104..4483
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4484..4687
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4638..4658
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4732..4751
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4732..4750
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4782..4791
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5319..5345
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(5348..5364)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5348..5364)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(5374..5392)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5375..5392)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5412..5429)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5418..5642
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(5688..5952,1..164)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5775..5794)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
       61 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
      121 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
      181 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
      241 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
      301 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
      361 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
      421 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
      481 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
      541 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
      601 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
      661 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
      721 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
      781 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
      841 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
      901 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
      961 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     1021 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     1081 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     1141 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     1201 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     1261 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     1321 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     1381 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     1441 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     1501 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     1561 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     1621 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     1681 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     1741 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     1801 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     1861 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     1921 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     1981 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2041 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2101 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2161 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2221 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2281 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2341 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2401 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2461 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2521 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2581 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2641 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2701 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2761 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     2821 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     2881 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     2941 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3001 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3061 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3121 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3181 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3241 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     3301 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     3361 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     3421 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     3481 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     3541 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3601 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     3661 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     3721 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     3781 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     3841 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     3901 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     3961 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     4021 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
     4081 tgtacgggcc agatatacgc gttgacattg attattgact agttattaat agtaatcaat
     4141 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     4201 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     4261 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     4321 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     4381 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     4441 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
     4501 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     4561 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     4621 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     4681 cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac
     4741 tcactatagg gagacccaag cttggtaccg agctcggatc tgccaccatg gggaaggtgc
     4801 tatccaaaat cttcgggaac aaggaaatgc ggatcctcat gttgggcctg gacgcggccg
     4861 gcaagacaac aatcctgtac aagttgaagc tgggccagtc ggtgaccacc attcccactg
     4921 tgggtttcaa cgtggagacg gtgacttaca aaaatgtcaa gttcaacgta tgggatgtgg
     4981 gcggcctgga caagatccgg ccgctctggc ggcattacta cactgggacc caaggtctca
     5041 tcttcgtagt ggactgcgcc gaccgcgacc gcatcgatga ggctcgccag gagctgcacc
     5101 gcattatcaa tgaccgggag atgagggacg ccataatcct catcttcgcc aacaagcagg
     5161 acctgcccga tgccatgaaa ccccacgaga tccaggagaa actgggcctg acccggattc
     5221 gggacaggaa ctggtatgtg cagccctcct gtgccacctc aggggacgga ctctatgagg
     5281 ggctcacatg gttaacctct aactacaaat ctctcgagta tccatacgat gtaccagact
     5341 acgcagagat ccactagttc tagagggccc tattctatag tgtcacctaa atgctagagc
     5401 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     5461 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     5521 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     5581 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     5641 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     5701 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     5761 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     5821 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     5881 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     5941 gacggttttt cg
//
