LOCUS       sequence-128537-        7283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of MKLP1 with an eGFP fusion and S708A/S710A 
            mutations.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7283)
  AUTHORS   Douglas ME, Davies T, Joseph N, Mishima M
  TITLE     Aurora B and 14-3-3 coordinately regulate clustering of 
            centralspindlin during cytokinesis.
  JOURNAL   Curr Biol. 2010 May 25;20(10):927-33. doi: 
            10.1016/j.cub.2010.03.055. Epub 2010 May 6.
  PUBMED    20451386
REFERENCE   2  (bases 1 to 7283)
  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: 
            "10.1016/j.cub.2010.03.055"; journalName: "Curr Biol"; date: 
            "2010-05-25- 25"; volume: "20"; issue: "10"; pages: "927-33"
COMMENT     Sequence Label: pEGFP-C1-MKLP1 S708/710A
FEATURES             Location/Qualifiers
     source          1..7283
                     /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             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4071..4192
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4108..4127)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4162..4181
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4199..4654)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4336..4357)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4548..4567
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4681..4785
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4787..5144
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4787..4978
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4810..4830)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4995..5130
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5179..5973
                     /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             5179..5973
                     /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(5233..5252)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5843..5862
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6161..6180)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6205..6252
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6581..7169
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7070..7089
                     /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 gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct ctgggcatgg ggaagtcagc gagagctaag
     1381 acaccccgga aacctaccgt gaaaaaaggg tcccaaacga accttaaaga cccagttggg
     1441 gtatactgta gggtgcgccc actgggcttt cctgatcaag agtgttgcat agaagtgatc
     1501 aataatacaa ctgttcagct tcatactcct gagggctaca gactcaaccg aaatggagac
     1561 tataaggaga ctcagtattc atttaaacaa gtatttggca ctcacaccac ccagaaggaa
     1621 ctctttgatg ttgtggctaa tcccttggtc aatgacctca ttcatggcaa aaatggtctt
     1681 ctttttacat atggtgtgac gggaagtgga aaaactcaca caatgactgg ttctccaggg
     1741 gaaggagggc tgcttcctcg ttgtttggac atgatcttta acagtatagg gtcatttcaa
     1801 gctaaacgat atgttttcaa atctaatgat aggaatagta tggatataca gtgtgaggtt
     1861 gatgccttat tagaacgtca gaaaagagaa gctatgccca atccaaagac ttcttctagc
     1921 aaacgacaag tagatccaga gtttgcagat atgataactg tacaagaatt ctgcaaagca
     1981 gaagaggttg atgaagatag tgtctatggt gtatttgtct cttatattga aatatataat
     2041 aattacatat atgatctatt ggaagaggtg ccgtttgatc ccataaaacc caaacctcca
     2101 caatctaaat tgcttcgtga agataagaac cataacatgt atgttgcagg atgtacagaa
     2161 gttgaagtga aatctactga ggaggctttt gaagttttct ggagaggcca gaaaaagaga
     2221 cgtattgcta atacccattt gaatcgtgag tccagccgtt cccatagcgt gttcaacatt
     2281 aaattagttc aggctccctt ggatgcagat ggagacaatg tcttacagga aaaagaacaa
     2341 atcactataa gtcagttgtc cttggtagat cttgctggaa gtgaaagaac taaccggacc
     2401 agagcagaag ggaacagatt acgtgaagct ggtaatatta atcagtcact aatgacgcta
     2461 agaacatgta tggatgtcct aagagagaac caaatgtatg gaactaacaa gatggttcca
     2521 tatcgagatt caaagttaac ccatctgttc aagaactact ttgatgggga aggaaaagtg
     2581 cggatgatcg tgtgtgtgaa ccccaaggct gaagattatg aagaaaactt gcaagtcatg
     2641 agatttgcgg aagtgactca agaagttgaa gtagcaagac ctgtcgacaa ggcaatatgt
     2701 ggtttaacgc ctgggaggag atacagaaac cagcctcgag gtccagttgg aaatgaacca
     2761 ttggttactg acgtggtttt gcagagtttt ccacctttgc cgtcatgcga aattttggat
     2821 atcaacgatg agcagacact tccaaggctg attgaagcct tagagaaacg acataactta
     2881 cgacaaatga tgattgatga gtttaacaaa caatctaatg cttttaaagc tttgttacaa
     2941 gaatttgaca atgctgtttt aagtaaagaa aaccacatgc aagggaaact aaatgaaaag
     3001 gagaagatga tctcaggaca gaaattggaa atagaacgac tggaaaagaa aaacaaaact
     3061 ttagaatata agattgagat tttagagaaa acaactacta tctatgagga agataaacgc
     3121 aatttgcaac aggaacttga aactcagaac cagaaacttc agcgacagtt ttctgacaaa
     3181 cgcagattag aagccaggtt gcaaggcatg gtgacagaaa cgacaatgaa gtgggagaaa
     3241 gaatgtgagc gtagagtggc agccaaacag ctggagatgc agaataaact ctgggttaaa
     3301 gatgaaaagc tgaaacaact gaaggctatt gttactgaac ctaaaactga gaagccagag
     3361 agaccctctc gggagcgaga tcgagaaaaa gttactcaaa gatctgtttc tccatcacct
     3421 gtgcctttac tctttcaacc tgatcagaac gcaccaccaa ttcgtctccg acacagacga
     3481 gcacgcgctg caggagacag atgggtagat cataagcccg cctctaacat gcaaactgaa
     3541 acagtcatgc agccacatgt ccctcatgcc atcacagtat ctgttgcaaa tgaaaaggca
     3601 ctagctaagt gtgagaagta catgctgacc caccaggaac tagcctccga tggggagatt
     3661 gaaactaaac taattaaggg tgatatttat aaaacaaggg gtggtggaca atctgttcag
     3721 tttactgata ttgagacttt aaagcaagaa tcaccaaatg gtagtcgaaa acgaagatct
     3781 tccacagtag cacctgccca accagatggt gcagagtctg aatggaccga tgtagaaaca
     3841 aggtgttctg tggctgtgga gatgagagca ggatcccagc tgggacctgg atatcagcat
     3901 catgcacaac ccaagcgcaa aaagccatga ccgcgggccc gggatccacc ggatctagat
     3961 aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     4021 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     4081 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     4141 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa
     4201 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     4261 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     4321 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     4381 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     4441 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     4501 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     4561 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     4621 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     4681 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4741 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     4801 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4861 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4921 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4981 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     5041 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     5101 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     5161 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     5221 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     5281 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     5341 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     5401 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     5461 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     5521 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     5581 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     5641 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     5701 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     5761 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     5821 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     5881 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     5941 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     6001 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     6061 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     6121 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     6181 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     6241 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     6301 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     6361 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     6421 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     6481 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6541 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6601 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6661 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6721 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6781 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6841 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6901 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6961 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     7021 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     7081 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     7141 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     7201 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     7261 ataaccgtat taccgccatg cat
//
