LOCUS       sequence-176230-        6656 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C-terminally mCherry-tagged Arf5(WT) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6656)
  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 6656)
  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/hArf5(WT)-mCherry
FEATURES             Location/Qualifiers
     source          1..6656
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(101..119)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(102..119)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(139..156)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    145..369
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      415..843
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(502..521)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     712..733
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(852..872)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        857..1186
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1037..1172
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1099..1118
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1253..2047
                     /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(1307..1326)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1917..1936
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2221..2342
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2258..2277)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2312..2331
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2391..2407)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2391..2407)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2404..2426)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2415..2431
                     /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(2439..2445,2446..2463,2464..2469))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2484..2505
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2622..2639)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2793..3381)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2873..2892)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3552..4343,4344..4412))
                     /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     4175..4194
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4413..4517)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4592..4611)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4783..5162
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5163..5366
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5317..5337
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5411..5430
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5411..5429
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      5461..5470
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      6025..6034
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6031..6656,1..85)
                     /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(6176..6194)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     6444..6463
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg
       61 cggcatggac gagctgtaca agtaatctag agggccctat tctatagtgt cacctaaatg
      121 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
      181 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
      241 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
      301 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
      361 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
      421 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
      481 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
      541 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
      601 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
      661 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
      721 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
      781 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
      841 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
      901 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
      961 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     1021 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     1081 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     1141 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     1201 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     1261 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     1321 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     1381 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     1441 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     1501 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     1561 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     1621 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     1681 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     1741 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     1801 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     1861 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     1921 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     1981 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     2041 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     2101 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     2161 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     2221 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     2281 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     2341 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     2401 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     2461 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     2521 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     2581 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     2641 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2701 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2761 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2821 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2881 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2941 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3001 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     3061 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3121 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3181 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3241 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     3301 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     3361 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     3421 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     3481 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3541 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3601 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3661 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3721 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3781 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3841 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     3901 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     3961 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4021 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4081 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4141 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4201 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4261 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4321 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4381 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4441 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4501 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     4561 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     4621 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
     4681 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
     4741 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta
     4801 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     4861 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     4921 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     4981 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     5041 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     5101 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     5161 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     5221 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     5281 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     5341 ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact gcttactggc
     5401 ttatcgaaat taatacgact cactataggg agacccaagc ttggtaccga gctcggatcc
     5461 gccaccatgg gcctcaccgt gtccgcgctc ttttcgcgga tcttcgggaa gaagcagatg
     5521 cggattctca tggttggctt ggatgcggct ggcaagacca caatcctgta caaactgaag
     5581 ttgggggaga ttgtcaccac catcccaacc ataggcttca atgtagaaac agtggaatat
     5641 aagaacatct gtttcacagt ctgggacgtg ggaggccagg acaagattcg gcctctgtgg
     5701 cggcactact tccagaacac tcagggcctc atctttgtgg tggacagtaa tgaccgggag
     5761 cgggtccaag aatctgctga tgaactccag aagatgctgc aggaggacga gctgcgggat
     5821 gcagtgctgc tggtatttgc caacaagcag gacatgccca acgccatgcc cgtgagcgag
     5881 ctgactgaca agctggggct acagcactta cgcagccgca cgtggtatgt ccaggccacc
     5941 tgtgccaccc aaggcacagg tctgtacgat ggtctggact ggctgtccca cgagctgtca
     6001 aagcgcctcg agggacttag gtctgccacc atggtgagca agggcgagga ggataacatg
     6061 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac
     6121 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
     6181 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
     6241 atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg
     6301 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
     6361 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc
     6421 ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
     6481 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg
     6541 aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag
     6601 cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctccca
//
