LOCUS       sequence-208758-        8372 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  inducible expression of vsv-Mis12-dCEN-INCENP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8372)
  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 8372)
  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: pcDNA4-Tet-on-vsv-Mis12-dCEN Incenp
FEATURES             Location/Qualifiers
     source          1..8372
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1005..1037
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     primer_bind     complement(4380..4397)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4386..4610
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4656..5084
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4743..4762)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4953..4974
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5093..5113)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5098..5427
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5278..5413
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5340..5359
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5475..5522
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5541..5915
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    6045..6166
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6082..6101)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6136..6155
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6215..6231)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6215..6231)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6228..6250)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6239..6255
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6263..6269,6270..6287,6288..6293))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6308..6329
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6446..6463)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6617..7205)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6697..6716)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7376..8167,8168..8236))
                     /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     7999..8018
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8237..8341)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtaccg gatccgccac catgtacacc gacatcgaga
     1021 tgaaccggtt gggcaaggaa ttctctgtgg atccaatgac ctacgaggcc cagttctttg
     1081 gcttcacgcc acaaacgtgc atgcttcgga tctacattgc atttcaagac tacctatttg
     1141 aagtgatgca ggccgttgaa caggttattc tgaagaagct ggatggcatc ccagactgtg
     1201 acattagccc agtgcagatt cgcaaatgca cagagaagtt tctttgcttc atgaaaggac
     1261 attttgataa cctttttagc aaaatggagc aactgttttt gcagctgatt ttacgtattc
     1321 cctcaaacat cttgcttcct gaagataaat gtaaggagac accttatagt gaggaagatt
     1381 ttcagcatct ccagaaagaa attgaacagt tacaggagaa gtacaagact gaattatgta
     1441 ctaagcaggc ccttcttgca gaattagaag agcaaaaaat tgttcaggcc aaactcaaac
     1501 agacgttgac tttctttgat gagcttcata atgttggcag agatcatggg actagtgatt
     1561 ttagggagag tttagtatcc ctggttcaga actccagaaa actacagaac attagagaca
     1621 atgtggaaaa ggaatcgaaa cgactgaaaa tatctgaatt cagcaaagag ccagagctga
     1681 tgcccaaaac accttctcag aagaaccgac ggaagaagag acggatttct tatgttcagg
     1741 atgaaaacag agatcccatc aggagaaggt tatcccgcag aaagtctcgg agcagccagc
     1801 tgagctcccg acgcctccgc agcaaggaca gtgtagagaa gctggctaca gtggtcgggg
     1861 agaacggctc cgttctgcgg cgtgtgaccc gtgctgcggc tgcagctgcc gcggctacca
     1921 tggcattggc tgcaccttct tcacccaccc ctgagtctcc cacgatgctg actaagaagc
     1981 ccgaggataa ccacacccag tgccagctgg tgcctgtggt ggagatcggc atcagtgagc
     2041 gccagaatgc tgagcagcat gtcacccagc tcatgtccac cgagcctctg ccccgcactc
     2101 tgtccccgac tccagcttca gccacagctc caacctccca gggcatcccg acatcagatg
     2161 aggaatcaac acctaagaag tcgaaggcca ggatactgga gtccatcaca gtgagctccc
     2221 tgatggctac accccaggac cccaagggtc aaggggtcgg gacggggcgg tctgcgtcta
     2281 agctcaggat tgcgcaggtc tcccctggcc cacgggactc gccagccttt ccagattctc
     2341 catggcggga gcgggtgctg gctcccatcc tgccggataa cttctccacg cccacgggct
     2401 ctcgcacgga ctctcaatcg gtgcggcaca gcccgatcgc cccgtcttcc ccgagtcccc
     2461 gagtcttagc ccagaagtac tctctggtgg ccaaacagga aagtgttgtc cgcagggcga
     2521 gcagaaggct tgccaagaag actgccgaag agccagctgc ctctggccgc atcatctgtc
     2581 acagttacct ggagaggctc ctgaatgttg aggtgcccca gaaagttggt tctgagcaga
     2641 aggaaccccc cgaggaggct gagcctgtgg cggcagctga gccagaggtc cctgagaaca
     2701 acggaaataa ctcgtggccc cacaatgaca cagagatcgc caacagcaca cccaacccga
     2761 agcctgcagc cagcagcccg gaaacaccct ctgcagggca gcaagaggcc aagacggacc
     2821 aagcagatgg acccagagag ccaccgcaga gtgccaggag gaagcgcagc tacaagcagg
     2881 ccgtgagtga gctggacgag gagcagcacc tggaggatga ggagctgcag ccccccagga
     2941 gcaagacccc ttcctcaccc tgcccagcca gcaaggtggt acggcccctc cggacctttc
     3001 tgcacacagt gcagaggaac cagatgctca cgaccccgac ctcagcccca cgcagcgtca
     3061 tgaagtcctt tattaagcgc aacactcccc tgcgcatgga ccccaagtgc agcttcgtcg
     3121 agaaggagcg gcagcgcctg gagaatctgc ggcggaagga ggaggccgag cagctgcgca
     3181 ggcagaaggt ggaggaggac aagcggcggc ggctggagga ggtgaagctg aagcgtgagg
     3241 aacgcctccg caaggtgctg caggcccgcg agcgggtgga gcagatgaag gaggagaaga
     3301 agaagcagat tgagcagaag tttgctcaga tcgacgagaa gactgagaag gccaaggagg
     3361 agcggctggc agaggagaag gccaagaaaa aggcggcggc caagaagatg gaggaggtgg
     3421 aagcacgcag gaagcaggaa gaggatgcac gtaggctcag gtggctgcag caggaggagg
     3481 aagagcggcg gcaccaagag ctgctgcaga agaagaagga agaggagcag gagcggctgc
     3541 ggaaggcggc cgaggctaag cggctggcag agcagcggga gcaggagcgg cgggagcagg
     3601 agcggcgcga gcaggagcgg cgcgagcagg agcggcggga gcaggagcgg cgcgagcagg
     3661 agcgacagct ggcagagcag gagcgtcggc gggagcagga gcggctccag gccgagaggg
     3721 agctgcagga gcgggagaag gccctgcggc tgcagaagga gcagctgcag agggaactgg
     3781 aggagaagaa gaagaaggaa gagcagcagc gtctggctga gcggcagctg caggaggagc
     3841 aagagaagaa agccaaggag gcagcagggg ccagcaaggc cctgaatgtg actgtggacg
     3901 tgcagtctcc agcttgtacc tcatatcaga tgactccgca agggcacagg gcccctccca
     3961 agatcaaccc agataactac gggatggatc tgaatagcga cgactccacc gatgatgagg
     4021 cccatccccg gaagcccatc cccacctggg cccgaggcac cccgctcagc caggctatca
     4081 ttcaccagta ctaccaccca ccgaaccttc tggagctctt tggaaccatt ctcccactgg
     4141 acttggagga tatcttcaag aagagcaagc cccgctatca caagcgcacc agctctgctg
     4201 tctggaactc accgcccctg cagggcgcca gggtccccag cagcctggcc tacagcctga
     4261 agaagcacaa gcttggtacc gagctcggat ccactagtcc agtgtggtgg aattctgcag
     4321 atatccagca cagtggcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc
     4381 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     4441 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     4501 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     4561 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     4621 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     4681 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4741 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4801 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4861 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4921 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4981 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     5041 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     5101 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5161 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     5221 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     5281 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     5341 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     5401 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     5461 tctgatcagc acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg
     5521 acaaggtgag gaactaaacc atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc
     5581 gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc cgggacttcg
     5641 tggaggacga cttcgccggt gtggtccggg acgacgtgac cctgttcatc agcgcggtcc
     5701 aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc
     5761 tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca
     5821 tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac ccggccggca
     5881 actgcgtgca cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca
     5941 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     6001 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     6061 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6121 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     6181 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     6241 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6301 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     6361 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6421 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6481 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6541 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6601 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6661 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6721 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6781 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6841 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6901 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6961 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7021 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     7081 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7141 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7201 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7261 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7321 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7381 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7441 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7501 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7561 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7621 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7681 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7741 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7801 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7861 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7921 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7981 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     8041 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     8101 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8161 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8221 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8281 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8341 gcacatttcc ccgaaaagtg ccacctgacg tc
//
