LOCUS       sequence-176231-        6641 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally mCherry-tagged Arf6(WT) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6641)
  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 6641)
  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)-mCherry
FEATURES             Location/Qualifiers
     source          1..6641
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      147..282
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     209..228
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             363..1157
                     /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(417..436)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1027..1046
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1331..1452
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1368..1387)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1422..1441
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1501..1517)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1501..1517)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1514..1536)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1525..1541
                     /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(1549..1555,1556..1573,1574..1579))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1594..1615
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1732..1749)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1903..2491)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1983..2002)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2662..3453,3454..3522))
                     /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     3285..3304
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3523..3627)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3702..3721)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3893..4272
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4273..4476
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4427..4447
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4521..4540
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4521..4539
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4571..4580
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      5120..5129
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5126..5836
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(5271..5289)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5539..5558
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        complement(5852..5870)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5853..5870)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5890..5907)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5896..6120
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6166..6594
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6253..6272)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6463..6484
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6603..6623)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6608..6641,1..296)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
       61 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
      121 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
      181 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
      241 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
      301 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
      361 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
      421 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
      481 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
      541 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
      601 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
      661 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
      721 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
      781 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
      841 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
      901 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
      961 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     1021 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     1081 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     1141 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     1201 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     1261 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     1321 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     1381 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     1441 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     1501 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     1561 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     1621 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     1681 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     1741 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     1801 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     1861 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1921 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     1981 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2041 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2101 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2161 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2221 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2281 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2341 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2401 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2461 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2521 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2581 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2641 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     2701 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     2761 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     2821 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     2881 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     2941 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3001 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3061 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3121 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3181 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3241 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     3301 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3361 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3421 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3481 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     3541 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3601 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     3661 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
     3721 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     3781 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     3841 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
     3901 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     3961 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     4021 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     4081 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     4141 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     4201 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     4261 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     4321 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     4381 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     4441 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
     4501 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc ttggtaccga
     4561 gctcggatct gccaccatgg ggaaggtgct atccaaaatc ttcgggaaca aggaaatgcg
     4621 gatcctcatg ttgggcctgg acgcggccgg caagacaaca atcctgtaca agttgaagct
     4681 gggccagtcg gtgaccacca ttcccactgt gggtttcaac gtggagacgg tgacttacaa
     4741 aaatgtcaag ttcaacgtat gggatgtggg cggccaggac aagatccggc cgctctggcg
     4801 gcattactac actgggaccc aaggtctcat cttcgtagtg gactgcgccg accgcgaccg
     4861 catcgatgag gctcgccagg agctgcaccg cattatcaat gaccgggaga tgagggacgc
     4921 cataatcctc atcttcgcca acaagcagga cctgcccgat gccatgaaac cccacgagat
     4981 ccaggagaaa ctgggcctga cccggattcg ggacaggaac tggtatgtgc agccctcctg
     5041 tgccacctca ggggacggac tctatgaggg gctcacatgg ttaacctcta actacaaatc
     5101 tctcgaggga cttaggtctg ccaccatggt gagcaagggc gaggaggata acatggccat
     5161 catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt
     5221 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
     5281 ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta
     5341 cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt
     5401 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt
     5461 gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac
     5521 caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc
     5581 cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct
     5641 gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt
     5701 gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga
     5761 ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga
     5821 cgagctgtac aagtaatcta gagggcccta ttctatagtg tcacctaaat gctagagctc
     5881 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     5941 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     6001 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     6061 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     6121 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     6181 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     6241 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     6301 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     6361 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     6421 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     6481 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     6541 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     6601 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc t
//
