LOCUS       sequence-205249-        8024 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of vsv-Mis12-dCEN-INCENP-EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8024)
  AUTHORS   Liu D, Vader G, Vromans MJ, Lampson MA, Lens SM
  TITLE     Sensing chromosome bi-orientation by spatial separation of aurora B 
            kinase from kinetochore substrates.
  JOURNAL   Science. 2009 Mar 6;323(5919):1350-3. doi: 10.1126/science.1167000. 
            Epub 2009 Jan 15.
  PUBMED    19150808
REFERENCE   2  (bases 1 to 8024)
  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: 
            "10.1126/science.1167000"; journalName: "Science"; date: "2009-03-6-
            6"; volume: "323"; issue: "5919"; pages: "1350-3"
COMMENT     Sequence Label: pEGFP-vsv-Mis12-dCEN-Incenp-EGFP
FEATURES             Location/Qualifiers
     source          1..8024
                     /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"
     CDS             649..681
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     regulatory      3964..3973
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3970..3972,3973..3975,3976..4689)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3970..3972,3973..3975,3976..4686)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4015..4036)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4276..4295)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4623..4644
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4812..4933
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4849..4868)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4903..4922
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4940..5395)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5077..5098)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5289..5308
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5422..5526
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5528..5885
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5528..5719
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5551..5571)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5736..5871
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5920..6714
                     /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             5920..6714
                     /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(5974..5993)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6584..6603
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6902..6921)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6946..6993
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7322..7910
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7811..7830
                     /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 ccggactcag atctcgagct caagcttggt accggatccg ccaccatgta caccgacatc
      661 gagatgaacc ggttgggcaa ggaattctct gtggatccaa tgacctacga ggcccagttc
      721 tttggcttca cgccacaaac gtgcatgctt cggatctaca ttgcatttca agactaccta
      781 tttgaagtga tgcaggccgt tgaacaggtt attctgaaga agctggatgg catcccagac
      841 tgtgacatta gcccagtgca gattcgcaaa tgcacagaga agtttctttg cttcatgaaa
      901 ggacattttg ataacctttt tagcaaaatg gagcaactgt ttttgcagct gattttacgt
      961 attccctcaa acatcttgct tcctgaagat aaatgtaagg agacacctta tagtgaggaa
     1021 gattttcagc atctccagaa agaaattgaa cagttacagg agaagtacaa gactgaatta
     1081 tgtactaagc aggcccttct tgcagaatta gaagagcaaa aaattgttca ggccaaactc
     1141 aaacagacgt tgactttctt tgatgagctt cataatgttg gcagagatca tgggactagt
     1201 gattttaggg agagtttagt atccctggtt cagaactcca gaaaactaca gaacattaga
     1261 gacaatgtgg aaaaggaatc gaaacgactg aaaatatctg aattcagcaa agagccagag
     1321 ctgatgccca aaacaccttc tcagaagaac cgacggaaga agagacggat ttcttatgtt
     1381 caggatgaaa acagagatcc catcaggaga aggttatccc gcagaaagtc tcggagcagc
     1441 cagcagagca cccgacgcct ccgcagcaag gacagtgtag agaagctggc tacagtggtc
     1501 ggggagaacg gctccgttct gcggcgtgtg acccgtgctg cggctgcagc tgccgcggct
     1561 accatggcat tggctgcacc ttcttcaccc acccctgagt ctcccacgat gctgactaag
     1621 aagcccgagg ataaccacac ccagtgccag ctggtgcctg tggtggagat cggcatcagt
     1681 gagcgccaga atgctgagca gcatgtcacc cagctcatgt ccaccgagcc tctgccccgc
     1741 actctgtccc cgactccagc ttcagccaca gctccaacct cccagggcat cccgacatca
     1801 gatgaggaat caacacctaa gaagtcgaag gccaggatac tggagtccat cacagtgagc
     1861 tccctgatgg ctacacccca ggaccccaag ggtcaagggg tcgggacggg gcggtctgcg
     1921 tctaagctca ggattgcgca ggtctcccct ggcccacggg actcgccagc ctttccagat
     1981 tctccatggc gggagcgggt gctggctccc atcctgccgg ataacttctc cacgcccacg
     2041 ggctctcgca cggactctca atcggtgcgg cacagcccga tcgccccgtc ttccccgagt
     2101 ccccgagtct tagcccagaa gtactctctg gtggccaaac aggaaagtgt tgtccgcagg
     2161 gcgagcagaa ggcttgccaa gaagactgcc gaagagccag ctgcctctgg ccgcatcatc
     2221 tgtcacagtt acctggagag gctcctgaat gttgaggtgc cccagaaagt tggttctgag
     2281 cagaaggaac cccccgagga ggctgagcct gtggcggcag ctgagccaga ggtccctgag
     2341 aacaacggaa ataactcgtg gccccacaat gacacagaga tcgccaacag cacacccaac
     2401 ccgaagcctg cagccagcag cccggaaaca ccctctgcag ggcagcaaga ggccaagacg
     2461 gaccaagcag atggacccag agagccaccg cagagtgcca ggaggaagcg cagctacaag
     2521 caggccgtga gtgagctgga cgaggagcag cacctggagg atgaggagct gcagcccccc
     2581 aggagcaaga ccccttcctc accctgccca gccagcaagg tggtacggcc cctccggacc
     2641 tttctgcaca cagtgcagag gaaccagatg ctcacgaccc cgacctcagc cccacgcagc
     2701 gtcatgaagt cctttattaa gcgcaacact cccctgcgca tggaccccaa gtgcagcttc
     2761 gtcgagaagg agcggcagcg cctggagaat ctgcggcgga aggaggaggc cgagcagctg
     2821 cgcaggcaga aggtggagga ggacaagcgg cggcggctgg aggaggtgaa gctgaagcgt
     2881 gaggaacgcc tccgcaaggt gctgcaggcc cgcgagcggg tggagcagat gaaggaggag
     2941 aagaagaagc agattgagca gaagtttgct cagatcgacg agaagactga gaaggccaag
     3001 gaggagcggc tggcagagga gaaggccaag aaaaaggcgg cggccaagaa gatggaggag
     3061 gtggaagcac gcaggaagca ggaagaggat gcacgtaggc tcaggtggct gcagcaggag
     3121 gaggaagagc ggcggcacca agagctgctg cagaagaaga aggaagagga gcaggagcgg
     3181 ctgcggaagg cggccgaggc taagcggctg gcagagcagc gggagcagga gcggcgggag
     3241 caggagcggc gcgagcagga gcggcgcgag caggagcggc gggagcagga gcggcgcgag
     3301 caggagcgac agctggcaga gcaggagcgt cggcgggagc aggagcggct ccaggccgag
     3361 agggagctgc aggagcggga gaaggccctg cggctgcaga aggagcagct gcagagggaa
     3421 ctggaggaga agaagaagaa ggaagagcag cagcgtctgg ctgagcggca gctgcaggag
     3481 gagcaagaga agaaagccaa ggaggcagta ggggccagca aggccctgaa tgtgactgtg
     3541 gacgtgcagt ctccagcttg tacctcatat caaatgactc cgcaagggca cagggcccct
     3601 cccaagatca acccagataa ctacgggatg gatctgaata gcgacgactc caccgatgat
     3661 gaggcccatc cccggaagcc catccccacc tgggcccgag gcaccccgct cagccaggct
     3721 atcattcacc agtactacca cccaccgaac cttctggagc tctttggaac cattctccca
     3781 ctggacttgg aggatatctt caagaagagc aagccccgct atcacaagcg caccagctct
     3841 gctgtctgga actcaccgcc cctgcagggc gccagggtcc ccagcagcct ggcctacagc
     3901 ctgaagaagc acaagcttcg aattctgcag tcgacggtac cgcgggcccg ggatccaccg
     3961 gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     4021 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     4081 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     4141 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     4201 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     4261 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     4321 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     4381 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     4441 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     4501 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     4561 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     4621 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     4681 tacaagtaaa gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt
     4741 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     4801 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     4861 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     4921 tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     4981 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     5041 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     5101 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     5161 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     5221 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     5281 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     5341 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     5401 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5461 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5521 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     5581 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     5641 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     5701 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     5761 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     5821 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     5881 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     5941 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     6001 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     6061 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     6121 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     6181 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     6241 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     6301 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     6361 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     6421 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     6481 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     6541 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     6601 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     6661 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     6721 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     6781 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     6841 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     6901 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     6961 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     7021 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     7081 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     7141 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     7201 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     7261 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     7321 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     7381 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     7441 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     7501 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     7561 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     7621 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     7681 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     7741 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     7801 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     7861 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     7921 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     7981 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
