LOCUS       sequence-208582-        8287 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of vsv-Mis12-dCEN-INCENP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8287)
  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 8287)
  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-Mis12-dCEN-Incenp
FEATURES             Location/Qualifiers
     source          1..8287
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        10..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     544..564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     638..657
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        638..656
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             691..723
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     promoter        complement(3996..4014)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3997..4014)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4034..4051)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4040..4264
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(4396..4984)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4476..4495)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5313..5360
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5385..5404
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5592..6386)
                     /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             complement(5592..6386)
                     /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(5703..5722)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6313..6332
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(6421..6778)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6435..6570
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6587..6778
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6735..6755
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(6838..7629,7630..7698))
                     /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     7461..7480
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7699..7803)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7830..8285
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7917..7936)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8127..8148
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gcgcgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
       61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      121 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      181 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
      601 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
      661 cccaagcttg gtaccggatc cgccaccatg tacaccgaca tcgagatgaa ccggttgggc
      721 aaggaattct ctgtggatcc aatgacctac gaggcccagt tctttggctt cacgccacaa
      781 acgtgcatgc ttcggatcta cattgcattt caagactacc tatttgaagt gatgcaggcc
      841 gttgaacagg ttattctgaa gaagctggat ggcatcccag actgtgacat tagcccagtg
      901 cagattcgca aatgcacaga gaagtttctt tgcttcatga aaggacattt tgataacctt
      961 tttagcaaaa tggagcaact gtttttgcag ctgattttac gtattccctc aaacatcttg
     1021 cttcctgaag ataaatgtaa ggagacacct tatagtgagg aagattttca gcatctccag
     1081 aaagaaattg aacagttaca ggagaagtac aagactgaat tatgtactaa gcaggccctt
     1141 cttgcagaat tagaagagca aaaaattgtt caggccaaac tcaaacagac gttgactttc
     1201 tttgatgagc ttcataatgt tggcagagat catgggacta gtgattttag ggagagttta
     1261 gtatccctgg ttcagaactc cagaaaacta cagaacatta gagacaatgt ggaaaaggaa
     1321 tcgaaacgac tgaaaatatc tgaattcagc aaagagccag agctgatgcc caaaacacct
     1381 tctcagaaga accgacggaa gaagagacgg atttcttatg ttcaggatga aaacagagat
     1441 cccatcagga gaaggttatc ccgcagaaag tctcggagca gccagctgag ctcccgacgc
     1501 ctccgcagca aggacagtgt agagaagctg gctacagtgg tcggggagaa cggctccgtt
     1561 ctgcggcgtg tgacccgtgc tgcggctgca gctgccgcgg ctaccatggc attggctgca
     1621 ccttcttcac ccacccctga gtctcccacg atgctgacta agaagcccga ggataaccac
     1681 acccagtgcc agctggtgcc tgtggtggag atcggcatca gtgagcgcca gaatgctgag
     1741 cagcatgtca cccagctcat gtccaccgag cctctgcccc gcactctgtc cccgactcca
     1801 gcttcagcca cagctccaac ctcccagggc atcccgacat cagatgagga atcaacacct
     1861 aagaagtcga aggccaggat actggagtcc atcacagtga gctccctgat ggctacaccc
     1921 caggacccca agggtcaagg ggtcgggacg gggcggtctg cgtctaagct caggattgcg
     1981 caggtctccc ctggcccacg ggactcgcca gcctttccag attctccatg gcgggagcgg
     2041 gtgctggctc ccatcctgcc ggataacttc tccacgccca cgggctctcg cacggactct
     2101 caatcggtgc ggcacagccc gatcgccccg tcttccccga gtccccgagt cttagcccag
     2161 aagtactctc tggtggccaa acaggaaagt gttgtccgca gggcgagcag aaggcttgcc
     2221 aagaagactg ccgaagagcc agctgcctct ggccgcatca tctgtcacag ttacctggag
     2281 aggctcctga atgttgaggt gccccagaaa gttggttctg agcagaagga accccccgag
     2341 gaggctgagc ctgtggcggc agctgagcca gaggtccctg agaacaacgg aaataactcg
     2401 tggccccaca atgacacaga gatcgccaac agcacaccca acccgaagcc tgcagccagc
     2461 agcccggaaa caccctctgc agggcagcaa gaggccaaga cggaccaagc agatggaccc
     2521 agagagccac cgcagagtgc caggaggaag cgcagctaca agcaggccgt gagtgagctg
     2581 gacgaggagc agcacctgga ggatgaggag ctgcagcccc ccaggagcaa gaccccttcc
     2641 tcaccctgcc cagccagcaa ggtggtacgg cccctccgga cctttctgca cacagtgcag
     2701 aggaaccaga tgctcacgac cccgacctca gccccacgca gcgtcatgaa gtcctttatt
     2761 aagcgcaaca ctcccctgcg catggacccc aagtgcagct tcgtcgagaa ggagcggcag
     2821 cgcctggaga atctgcggcg gaaggaggag gccgagcagc tgcgcaggca gaaggtggag
     2881 gaggacaagc ggcggcggct ggaggaggtg aagctgaagc gtgaggaacg cctccgcaag
     2941 gtgctgcagg cccgcgagcg ggtggagcag atgaaggagg agaagaagaa gcagattgag
     3001 cagaagtttg ctcagatcga cgagaagact gagaaggcca aggaggagcg gctggcagag
     3061 gagaaggcca agaaaaaggc ggcggccaag aagatggagg aggtggaagc acgcaggaag
     3121 caggaagagg atgcacgtag gctcaggtgg ctgcagcagg aggaggaaga gcggcggcac
     3181 caagagctgc tgcagaagaa gaaggaagag gagcaggagc ggctgcggaa ggcggccgag
     3241 gctaagcggc tggcagagca gcgggagcag gagcggcggg agcaggagcg gcgcgagcag
     3301 gagcggcgcg agcaggagcg gcgggagcag gagcggcgcg agcaggagcg acagctggca
     3361 gagcaggagc gtcggcggga gcaggagcgg ctccaggccg agagggagct gcaggagcgg
     3421 gagaaggccc tgcggctgca gaaggagcag ctgcagaggg aactggagga gaagaagaag
     3481 aaggaagagc agcagcgtct ggctgagcgg cagctgcagg aggagcaaga gaagaaagcc
     3541 aaggaggcag caggggccag caaggccctg aatgtgactg tggacgtgca gtctccagct
     3601 tgtacctcat atcagatgac tccgcaaggg cacagggccc ctcccaagat caacccagat
     3661 aactacggga tggatctgaa tagcgacgac tccaccgatg atgaggccca tccccggaag
     3721 cccatcccca cctgggcccg aggcaccccg ctcagccagg ctatcattca ccagtactac
     3781 cacccaccga accttctgga gctctttgga accattctcc cactggactt ggaggatatc
     3841 ttcaagaaga gcaagccccg ctatcacaag cgcaccagct ctgctgtctg gaactcaccg
     3901 cccctgcagg gcgccagggt ccccagcagc ctggcctaca gcctgaagaa gcacaagctt
     3961 tgactcgagt agatgactag tctagagggc cctattctat agtgtcacct aaatgctaga
     4021 gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     4081 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     4141 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     4201 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     4261 atggcttctg aggcggaaag aaccagtggc ggtaatacgg ttatccacag aatcagggga
     4321 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4381 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4441 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4501 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4561 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4621 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4681 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4741 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4801 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4861 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4921 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4981 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5041 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5101 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa cctgaggcta tggcagggcc
     5161 tgccgccccg acgttggctg cgagccctgg gccttcaccc gaacttgggg ggtggggtgg
     5221 ggaaaaggaa gaaacgcggg cgtattggcc ccaatggggt ctcggtgggg tatcgacaga
     5281 gtgccagccc tgggaccgaa ccccgcgttt atgaacaaac gacccaacac cgtgcgtttt
     5341 attctgtctt tttattgccg tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt
     5401 ttcagttagc ctccccctag ggtgggcgaa gaactccagc atgagatccc cgcgctggag
     5461 gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc
     5521 ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc
     5581 cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg
     5641 ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca
     5701 gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca
     5761 cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg
     5821 ccatgggtca cgacgagatc ctcgccgtcg ggcatgctcg ccttgagcct ggcgaacagt
     5881 tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct
     5941 tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta
     6001 gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca
     6061 ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc
     6121 cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc
     6181 cacgatagcc gcgctgcctc gtcttgcagt tcattcaggg caccggacag gtcggtcttg
     6241 acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg
     6301 attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct
     6361 gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcgatc
     6421 tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc
     6481 cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg
     6541 ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc
     6601 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     6661 acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     6721 tggggacttt ccacacccta actgacacac attccacagc tggttctttc cgcctcagga
     6781 ctcttccttt ttcaataaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6841 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6901 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6961 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7021 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7081 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7141 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7201 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7261 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7321 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7381 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7441 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7501 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7561 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7621 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7681 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7741 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7801 gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc
     7861 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     7921 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     7981 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     8041 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     8101 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     8161 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     8221 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     8281 aatttac
//
