LOCUS       sequence-128546-        7274 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of MKLP1 with an mCherry fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7274)
  AUTHORS   Joseph N, Hutterer A, Poser I, Mishima M
  TITLE     ARF6 GTPase protects the post-mitotic midbody from 14-3-3-mediated 
            disintegration.
  JOURNAL   EMBO J. 2012 May 30;31(11):2604-14. doi: 10.1038/emboj.2012.139. 
            Epub 2012 May 11.
  PUBMED    22580824
REFERENCE   2  (bases 1 to 7274)
  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.2012.139"; journalName: "EMBO J"; date: "2012-05-30- 
            30"; volume: "31"; issue: "11"; pages: "2604-14"
COMMENT     Sequence Label: pmCherry-C1-MKLP1
FEATURES             Location/Qualifiers
     source          1..7274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..1320
                     /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(758..776)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1026..1045
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4062..4183
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4099..4118)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4153..4172
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4190..4645)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4327..4348)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4539..4558
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4672..4776
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4778..5135
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4778..4969
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4801..4821)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4986..5121
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5170..5964
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5170..5964
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5224..5243)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5834..5853
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6152..6171)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6196..6243
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6572..7160
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7061..7080
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
      661 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
      721 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
      781 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
      841 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
      901 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
      961 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     1021 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     1081 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     1141 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     1201 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     1261 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     1321 tccggactca gatctcgagc tctgggcatg gggaagtcag cgagagctaa gacaccccgg
     1381 aaacctaccg tgaaaaaagg gtcccaaacg aaccttaaag acccagttgg ggtatactgt
     1441 agggtgcgcc cactgggctt tcctgatcaa gagtgttgca tagaagtgat caataataca
     1501 actgttcagc ttcatactcc tgagggctac agactcaacc gaaatggaga ctataaggag
     1561 actcagtatt catttaaaca agtatttggc actcacacca cccagaagga actctttgat
     1621 gttgtggcta atcccttggt caatgacctc attcatggca aaaatggtct tctttttaca
     1681 tatggtgtga cgggaagtgg aaaaactcac acaatgactg gttctccagg ggaaggaggg
     1741 ctgcttcctc gttgtttgga catgatcttt aacagtatag ggtcatttca agctaaacga
     1801 tatgttttca aatctaatga taggaatagt atggatatac agtgtgaggt tgatgcctta
     1861 ttagaacgtc agaaaagaga agctatgccc aatccaaaga cttcttctag caaacgacaa
     1921 gtagatccag agtttgcaga tatgataact gtacaagaat tctgcaaagc agaagaggtt
     1981 gatgaagata gtgtctatgg tgtatttgtc tcttatattg aaatatataa taattacata
     2041 tatgatctat tggaagaggt gccgtttgat cccataaaac ccaaacctcc acaatctaaa
     2101 ttgcttcgtg aagataagaa ccataacatg tatgttgcag gatgtacaga agttgaagtg
     2161 aaatctactg aggaggcttt tgaagttttc tggagaggcc agaaaaagag acgtattgct
     2221 aatacccatt tgaatcgtga gtccagccgt tcccatagcg tgttcaacat taaattagtt
     2281 caggctccct tggatgcaga tggagacaat gtcttacagg aaaaagaaca aatcactata
     2341 agtcagttgt ccttggtaga tcttgctgga agtgaaagaa ctaaccggac cagagcagaa
     2401 gggaacagat tacgtgaagc tggtaatatt aatcagtcac taatgacgct aagaacatgt
     2461 atggatgtcc taagagagaa ccaaatgtat ggaactaaca agatggttcc atatcgagat
     2521 tcaaagttaa cccatctgtt caagaactac tttgatgggg aaggaaaagt gcggatgatc
     2581 gtgtgtgtga accccaaggc tgaagattat gaagaaaact tgcaagtcat gagatttgcg
     2641 gaagtgactc aagaagttga agtagcaaga cctgtcgaca aggcaatatg tggtttaacg
     2701 cctgggagga gatacagaaa ccagcctcga ggtccagttg gaaatgaacc attggttact
     2761 gacgtggttt tgcagagttt tccacctttg ccgtcatgcg aaattttgga tatcaacgat
     2821 gagcagacac ttccaaggct gattgaagcc ttagagaaac gacataactt acgacaaatg
     2881 atgattgatg agtttaacaa acaatctaat gcttttaaag ctttgttaca agaatttgac
     2941 aatgctgttt taagtaaaga aaaccacatg caagggaaac taaatgaaaa ggagaagatg
     3001 atctcaggac agaaattgga aatagaacga ctggaaaaga aaaacaaaac tttagaatat
     3061 aagattgaga ttttagagaa aacaactact atctatgagg aagataaacg caatttgcaa
     3121 caggaacttg aaactcagaa ccagaaactt cagcgacagt tttctgacaa acgcagatta
     3181 gaagccaggt tgcaaggcat ggtgacagaa acgacaatga agtgggagaa agaatgtgag
     3241 cgtagagtgg cagccaaaca gctggagatg cagaataaac tctgggttaa agatgaaaag
     3301 ctgaaacaac tgaaggctat tgttactgaa cctaaaactg agaagccaga gagaccctct
     3361 cgggagcgag atcgagaaaa agttactcaa agatctgttt ctccatcacc tgtgccttta
     3421 ctctttcaac ctgatcagaa cgcaccacca attcgtctcc gacacagacg atcacgctct
     3481 gcaggagaca gatgggtaga tcataagccc gcctctaaca tgcaaactga aacagtcatg
     3541 cagccacatg tccctcatgc catcacagta tctgttgcaa atgaaaaggc actagctaag
     3601 tgtgagaagt acatgctgac ccaccaggaa ctagcctccg atggggagat tgaaactaaa
     3661 ctaattaagg gtgatattta taaaacaagg ggtggtggac aatctgttca gtttactgat
     3721 attgagactt taaagcaaga atcaccaaat ggtagtcgaa aacgaagatc ttccacagta
     3781 gcacctgccc aaccagatgg tgcagagtct gaatggaccg atgtagaaac aaggtgttct
     3841 gtggctgtgg agatgagagc aggatcccag ctgggacctg gatatcagca tcatgcacaa
     3901 cccaagcgca aaaagccatg accgcgggcc cgggatccac cggatctaga taactgatca
     3961 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     4021 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     4081 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4141 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc
     4201 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     4261 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     4321 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     4381 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     4441 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     4501 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     4561 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     4621 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4681 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4741 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     4801 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4861 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4921 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4981 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     5041 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     5101 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     5161 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     5221 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     5281 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     5341 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     5401 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     5461 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     5521 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     5581 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     5641 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     5701 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5761 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5821 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5881 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5941 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     6001 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     6061 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     6121 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     6181 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     6241 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     6301 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     6361 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     6421 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     6481 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6541 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6601 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6661 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     6721 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6781 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6841 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6901 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6961 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     7021 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     7081 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     7141 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     7201 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     7261 ttaccgccat gcat
//
