LOCUS       sequence-220770-        7910 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  inducible expression of vsv-Mis12-dCEN-INCENP-EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7910)
  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 7910)
  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: pCR3-vsv-CenpB 1-166-dCEN-Incenp-GFP
FEATURES             Location/Qualifiers
     source          1..7910
                     /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      3850..3859
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3856..3858,3859..3861,3862..4575)
                     /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(3856..3858,3859..3861,3862..4572)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3901..3922)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4162..4181)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4509..4530
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4698..4819
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4735..4754)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4789..4808
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4826..5281)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4963..4984)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5175..5194
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5308..5412
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5414..5771
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5414..5605
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5437..5457)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5622..5757
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5806..6600
                     /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             5806..6600
                     /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(5860..5879)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6470..6489
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6788..6807)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6832..6879
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7208..7796
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7697..7716
                     /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 ggaattcggc cccaagaggc gacagctgac gttccgggag
      721 aagtcacgga tcatccagga ggtggaggag aatccggacc tgcgcaaggg cgagatcgcg
      781 cggcgcttca acatcccgcc gtccacgctg agcacgatcc tgaagaacaa gcgcgccatc
      841 ctggcgtcgg agcgcaagta cggggtggcc tccacctgcc gcaagaccaa caagctgtct
      901 ccctacgaca agctcgaggg cttgctcatc gcctggttcc agcagatccg cgccgccggc
      961 ctgccggtca agggcatcat cctcaaggag aaggcgctgc gcatagccga ggagctgggc
     1021 atggacgact tcaccgcctc caacggctgg ctggaccgct tccgccggcg ccacggcgtg
     1081 gtgtcctgca gcggcgtggc ccgcgcccgc gcgcgaaacg ctgccccccg caccccggcg
     1141 gcgcctgcca gtccggccgc ggtgccctcg gagggcggga gcactgaatt cagcaaagag
     1201 ccagagctga tgcccaaaac accttctcag aagaaccgac ggaagaagag acggatttct
     1261 tatgttcagg atgaaaacag agatcccatc aggagaaggt tatcccgcag aaagtctcgg
     1321 agcagccagc tgagctcccg acgcctccgc agcaaggaca gtgtagagaa gctggctaca
     1381 gtggtcgggg agaacggctc cgttctgcgg cgtgtgaccc gtgctgcggc tgcagctgcc
     1441 gcggctacca tggcattggc tgcaccttct tcacccaccc ctgagtctcc cacgatgctg
     1501 actaagaagc ccgaggataa ccacacccag tgccagctgg tgcctgtggt ggagatcggc
     1561 atcagtgagc gccagaatgc tgagcagcat gtcacccagc tcatgtccac cgagcctctg
     1621 ccccgcactc tgtccccgac tccagcttca gccacagctc caacctccca gggcatcccg
     1681 acatcagatg aggaatcaac acctaagaag tcgaaggcca ggatactgga gtccatcaca
     1741 gtgagctccc tgatggctac accccaggac cccaagggtc aaggggtcgg gacggggcgg
     1801 tctgcgtcta agctcaggat tgcgcaggtc tcccctggcc cacgggactc gccagccttt
     1861 ccagattctc catggcggga gcgggtgctg gctcccatcc tgccggataa cttctccacg
     1921 cccacgggct ctcgcacgga ctctcaatcg gtgcggcaca gcccgatcgc cccgtcttcc
     1981 ccgagtcccc gagtcttagc ccagaagtac tctctggtgg ccaaacagga aagtgttgtc
     2041 cgcagggcga gcagaaggct tgccaagaag actgccgaag agccagctgc ctctggccgc
     2101 atcatctgtc acagttacct ggagaggctc ctgaatgttg aggtgcccca gaaagttggt
     2161 tctgagcaga aggaaccccc cgaggaggct gagcctgtgg cggcagctga gccagaggtc
     2221 cctgagaaca acggaaataa ctcgtggccc cacaatgaca cagagatcgc caacagcaca
     2281 cccaacccga agcctgcagc cagcagcccg gaaacaccct ctgcagggca gcaagaggcc
     2341 aagacggacc aagcagatgg acccagagag ccaccgcaga gtgccaggag gaagcgcagc
     2401 tacaagcagg ccgtgagtga gctggacgag gagcagcacc tggaggatga ggagctgcag
     2461 ccccccagga gcaagacccc ttcctcaccc tgcccagcca gcaaggtggt acggcccctc
     2521 cggacctttc tgcacacagt gcagaggaac cagatgctca cgaccccgac ctcagcccca
     2581 cgcagcgtca tgaagtcctt tattaagcgc aacactcccc tgcgcatgga ccccaagtgc
     2641 agcttcgtcg agaaggagcg gcagcgcctg gagaatctgc ggcggaagga ggaggccgag
     2701 cagctgcgca ggcagaaggt ggaggaggac aagcggcggc ggctggagga ggtgaagctg
     2761 aagcgtgagg aacgcctccg caaggtgctg caggcccgcg agcgggtgga gcagatgaag
     2821 gaggagaaga agaagcagat tgagcagaag tttgctcaga tcgacgagaa gactgagaag
     2881 gccaaggagg agcggctggc agaggagaag gccaagaaaa aggcggcggc caagaagatg
     2941 gaggaggtgg aagcacgcag gaagcaggaa gaggatgcac gtaggctcag gtggctgcag
     3001 caggaggagg aagagcggcg gcaccaagag ctgctgcaga agaagaagga agaggagcag
     3061 gagcggctgc ggaaggcggc cgaggctaag cggctggcag agcagcggga gcaggagcgg
     3121 cgggagcagg agcggcgcga gcaggagcgg cgcgagcagg agcggcggga gcaggagcgg
     3181 cgcgagcagg agcgacagct ggcagagcag gagcgtcggc gggagcagga gcggctccag
     3241 gccgagaggg agctgcagga gcgggagaag gccctgcggc tgcagaagga gcagctgcag
     3301 agggaactgg aggagaagaa gaagaaggaa gagcagcagc gtctggctga gcggcagctg
     3361 caggaggagc aagagaagaa agccaaggag gcagcagggg ccagcaaggc cctgaatgtg
     3421 actgtggacg tgcagtctcc agcttgtacc tcatatcaga tgactccgca agggcacagg
     3481 gcccctccca agatcaaccc agataactac gggatggatc tgaatagcga cgactccacc
     3541 gatgatgagg cccatccccg gaagcccatc cccacctggg cccgaggcac cccgctcagc
     3601 caggctatca ttcaccagta ctaccaccca ccgaaccttc tggagctctt tggaaccatt
     3661 ctcccactgg acttggagga tatcttcaag aagagcaagc cccgctatca caagcgcacc
     3721 agctctgctg tctggaactc accgcccctg cagggcgcca gggtccccag cagcctggcc
     3781 tacagcctga agaagcacaa gcttcgaatt ctgcagtcga cggtaccgcg ggcccgggat
     3841 ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     3901 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     3961 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     4021 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     4081 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     4141 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     4201 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     4261 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     4321 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     4381 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     4441 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     4501 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     4561 gagctgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     4621 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     4681 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4741 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4801 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     4861 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     4921 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     4981 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     5041 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     5101 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     5161 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     5221 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     5281 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5341 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5401 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     5461 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5521 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5581 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     5641 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     5701 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     5761 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     5821 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     5881 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     5941 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     6001 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     6061 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     6121 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     6181 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     6241 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     6301 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     6361 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     6421 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     6481 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     6541 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     6601 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     6661 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     6721 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     6781 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     6841 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     6901 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     6961 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     7021 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     7081 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     7141 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     7201 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     7261 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     7321 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     7381 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7441 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7501 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7561 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7621 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7681 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7741 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7801 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     7861 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
