LOCUS       sequence-208751-        8193 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  inducible expression of vsv-CenpB-1-166-dCEN-INCENP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8193)
  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 8193)
  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-CenpB 1-166-dCEN Incenp
FEATURES             Location/Qualifiers
     source          1..8193
                     /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(4201..4218)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4207..4431
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4477..4905
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4564..4583)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4774..4795
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4914..4934)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4919..5248
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5099..5234
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5161..5180
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5296..5343
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5362..5736
                     /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    5866..5987
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5903..5922)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5957..5976
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6036..6052)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6036..6052)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6049..6071)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6060..6076
                     /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(6084..6090,6091..6108,6109..6114))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6129..6150
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6267..6284)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6438..7026)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6518..6537)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7197..7988,7989..8057))
                     /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     7820..7839
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8058..8162)
                     /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 ttcggcccca agaggcgaca gctgacgttc cgggagaagt
     1081 cacggatcat ccaggaggtg gaggagaatc cggacctgcg caagggcgag atcgcgcggc
     1141 gcttcaacat cccgccgtcc acgctgagca cgatcctgaa gaacaagcgc gccatcctgg
     1201 cgtcggagcg caagtacggg gtggcctcca cctgccgcaa gaccaacaag ctgtctccct
     1261 acgacaagct cgagggcttg ctcatcgcct ggttccagca gatccgcgcc gccggcctgc
     1321 cggtcaaggg catcatcctc aaggagaagg cgctgcgcat agccgaggag ctgggcatgg
     1381 acgacttcac cgcctccaac ggctggctgg accgcttccg ccggcgccac ggcgtggtgt
     1441 cctgcagcgg cgtggcccgc gcccgcgcgc gaaacgctgc cccccgcacc ccggcggcgc
     1501 ctgccagtcc ggccgcggtg ccctcggagg gcgggagcac tgaattcagc aaagagccag
     1561 agctgatgcc caaaacacct tctcagaaga accgacggaa gaagagacgg atttcttatg
     1621 ttcaggatga aaacagagat cccatcagga gaaggttatc ccgcagaaag tctcggagca
     1681 gccagctgag ctcccgacgc ctccgcagca aggacagtgt agagaagctg gctacagtgg
     1741 tcggggagaa cggctccgtt ctgcggcgtg tgacccgtgc tgcggctgca gctgccgcgg
     1801 ctaccatggc attggctgca ccttcttcac ccacccctga gtctcccacg atgctgacta
     1861 agaagcccga ggataaccac acccagtgcc agctggtgcc tgtggtggag atcggcatca
     1921 gtgagcgcca gaatgctgag cagcatgtca cccagctcat gtccaccgag cctctgcccc
     1981 gcactctgtc cccgactcca gcttcagcca cagctccaac ctcccagggc atcccgacat
     2041 cagatgagga atcaacacct aagaagtcga aggccaggat actggagtcc atcacagtga
     2101 gctccctgat ggctacaccc caggacccca agggtcaagg ggtcgggacg gggcggtctg
     2161 cgtctaagct caggattgcg caggtctccc ctggcccacg ggactcgcca gcctttccag
     2221 attctccatg gcgggagcgg gtgctggctc ccatcctgcc ggataacttc tccacgccca
     2281 cgggctctcg cacggactct caatcggtgc ggcacagccc gatcgccccg tcttccccga
     2341 gtccccgagt cttagcccag aagtactctc tggtggccaa acaggaaagt gttgtccgca
     2401 gggcgagcag aaggcttgcc aagaagactg ccgaagagcc agctgcctct ggccgcatca
     2461 tctgtcacag ttacctggag aggctcctga atgttgaggt gccccagaaa gttggttctg
     2521 agcagaagga accccccgag gaggctgagc ctgtggcggc agctgagcca gaggtccctg
     2581 agaacaacgg aaataactcg tggccccaca atgacacaga gatcgccaac agcacaccca
     2641 acccgaagcc tgcagccagc agcccggaaa caccctctgc agggcagcaa gaggccaaga
     2701 cggaccaagc agatggaccc agagagccac cgcagagtgc caggaggaag cgcagctaca
     2761 agcaggccgt gagtgagctg gacgaggagc agcacctgga ggatgaggag ctgcagcccc
     2821 ccaggagcaa gaccccttcc tcaccctgcc cagccagcaa ggtggtacgg cccctccgga
     2881 cctttctgca cacagtgcag aggaaccaga tgctcacgac cccgacctca gccccacgca
     2941 gcgtcatgaa gtcctttatt aagcgcaaca ctcccctgcg catggacccc aagtgcagct
     3001 tcgtcgagaa ggagcggcag cgcctggaga atctgcggcg gaaggaggag gccgagcagc
     3061 tgcgcaggca gaaggtggag gaggacaagc ggcggcggct ggaggaggtg aagctgaagc
     3121 gtgaggaacg cctccgcaag gtgctgcagg cccgcgagcg ggtggagcag atgaaggagg
     3181 agaagaagaa gcagattgag cagaagtttg ctcagatcga cgagaagact gagaaggcca
     3241 aggaggagcg gctggcagag gagaaggcca agaaaaaggc ggcggccaag aagatggagg
     3301 aggtggaagc acgcaggaag caggaagagg atgcacgtag gctcaggtgg ctgcagcagg
     3361 aggaggaaga gcggcggcac caagagctgc tgcagaagaa gaaggaagag gagcaggagc
     3421 ggctgcggaa ggcggccgag gctaagcggc tggcagagca gcgggagcag gagcggcggg
     3481 agcaggagcg gcgcgagcag gagcggcgcg agcaggagcg gcgggagcag gagcggcgcg
     3541 agcaggagcg acagctggca gagcaggagc gtcggcggga gcaggagcgg ctccaggccg
     3601 agagggagct gcaggagcgg gagaaggccc tgcggctgca gaaggagcag ctgcagaggg
     3661 aactggagga gaagaagaag aaggaagagc agcagcgtct ggctgagcgg cagctgcagg
     3721 aggagcaaga gaagaaagcc aaggaggcag caggggccag caaggccctg aatgtgactg
     3781 tggacgtgca gtctccagct tgtacctcat atcagatgac tccgcaaggg cacagggccc
     3841 ctcccaagat caacccagat aactacggga tggatctgaa tagcgacgac tccaccgatg
     3901 atgaggccca tccccggaag cccatcccca cctgggcccg aggcaccccg ctcagccagg
     3961 ctatcattca ccagtactac cacccaccga accttctgga gctctttgga accattctcc
     4021 cactggactt ggaggatatc ttcaagaaga gcaagccccg ctatcacaag cgcaccagct
     4081 ctgctgtctg gaactcaccg cccctgcagg gcgccagggt ccccagcagc ctggcctaca
     4141 gcctgaagaa gcacaagctt tgactcgagt ctagagggcc cgtttaaacc cgctgatcag
     4201 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     4261 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     4321 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     4381 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     4441 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     4501 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     4561 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4621 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4681 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4741 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4801 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     4861 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     4921 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4981 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     5041 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     5101 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     5161 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     5221 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     5281 atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac
     5341 gacaaggtga ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg
     5401 cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc
     5461 gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc
     5521 caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag
     5581 ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc
     5641 atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc
     5701 aactgcgtgc acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc
     5761 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5821 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     5881 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5941 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     6001 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     6061 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     6121 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6181 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6241 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6301 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6361 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6421 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6481 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6541 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6601 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6661 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6721 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6781 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6841 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6901 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6961 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7021 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7081 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7141 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7201 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7261 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7321 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7381 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7441 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7501 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7561 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7621 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7681 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7741 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7801 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7861 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7921 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7981 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     8041 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     8101 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     8161 cgcacatttc cccgaaaagt gccacctgac gtc
//
