LOCUS       sequence-208576-        7568 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of vsv-INCENP d543-746.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7568)
  AUTHORS   Vader G, Cruijsen CW, van Harn T, Vromans MJ, Medema RH, Lens SM
  TITLE     The chromosomal passenger complex controls spindle checkpoint 
            function independent from its role in correcting microtubule 
            kinetochore interactions.
  JOURNAL   Mol Biol Cell. 2007 Nov;18(11):4553-64. doi: 
            10.1091/mbc.E07-04-0328. Epub 2007 Aug 15.
  PUBMED    17699588
REFERENCE   2  (bases 1 to 7568)
  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: 
            "10.1091/mbc.E07-04-0328"; journalName: "Mol Biol Cell"; date: 
            "2007-11"; volume: "18"; issue: "11"; pages: "4553-64"
COMMENT     Sequence Label: pCR3-vsv-INCENP d543-746
FEATURES             Location/Qualifiers
     source          1..7568
                     /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"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             910..942
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     promoter        complement(3121..3139)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3122..3139)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3159..3176)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3165..3389
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3435..3863
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3522..3541)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3732..3753
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3872..3892)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3877..4206
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4057..4192
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4119..4138
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4273..5067
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4327..4346)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4937..4956
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5241..5362
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5278..5297)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5332..5351
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5411..5427)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5411..5427)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5424..5446)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5435..5451
                     /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(5459..5465,5466..5483,5484..5489))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5504..5525
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5642..5659)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5813..6401)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5893..5912)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6572..7363,7364..7432))
                     /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     7195..7214
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7433..7537)
                     /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 ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gccaccatgt acaccgacat cgagatgaac cggttgggca aggggacgac ggccccaggg
      961 cccattcacc tgctggagct ctgtgaccag aagctcatgg agtttctctg caacatggat
     1021 aataaggact tggtgtggct tgaggagatc caagaggagg ccgagcgcat gttcaccaga
     1081 gaattcagca aagagccaga gctgatgccc aaaacacctt ctcagaagaa ccgacggaag
     1141 aagagacgga tttcttatgt tcaggatgaa aacagagatc ccatcaggag aaggttatcc
     1201 cgcagaaagt ctcggagcag ccagctgagc tcccgacgcc tccgcagcaa ggacagtgta
     1261 gagaagctgg ctacagtggt cggggagaac ggctccgttc tgcggcgtgt gacccgtgct
     1321 gcggctgcag ctgccgcggc taccatggca ttggctgcac cttcttcacc cacccctgag
     1381 tctcccacga tgctgactaa gaagcccgag gataaccaca cccagtgcca gctggtgcct
     1441 gtggtggaga tcggcatcag tgagcgccag aatgctgagc agcatgtcac ccagctcatg
     1501 tccaccgagc ctctgccccg cactctgtcc ccgactccag cttcagccac agctccaacc
     1561 tcccagggca tcccgacatc agatgaggaa tcaacaccta agaagtcgaa ggccaggata
     1621 ctggagtcca tcacagtgag ctccctgatg gctacacccc aggaccccaa gggtcaaggg
     1681 gtcgggacgg ggcggtctgc gtctaagctc aggattgcgc aggtctcccc tggcccacgg
     1741 gactcgccag cctttccaga ttctccatgg cgggagcggg tgctggctcc catcctgccg
     1801 gataacttct ccacgcccac gggctctcgc acggactctc aatcggtgcg gcacagcccg
     1861 atcgccccgt cttccccgag tccccgagtc ttagcccaga agtactctct ggtggccaaa
     1921 caggaaagtg ttgtccgcag ggcgagcaga aggcttgcca agaagactgc cgaagagcca
     1981 gctgcctctg gccgcatcat ctgtcacagt tacctggaga ggctcctgaa tgttgaggtg
     2041 ccccagaaag ttggttctga gcagaaggaa ccccccgagg aggctgagcc tgtggcggca
     2101 gctgagccag aggtccctga gaacaacgga aataactcgt ggccccacaa tgacacagag
     2161 atcgccaaca gcacacccaa cccgaagcct gcagccagca gcccggaaac accctctgca
     2221 gggcagcaag aggccaagac ggaccaagca gatggaccca gagagccacc gcagagtgcc
     2281 aggaggaagc gcagctacaa gcaggccgtg agtgagctgg acgaggagca gcacctggag
     2341 gatgaggagc tgcagccccc caggagcaag accccttcct caccctgccc agccagcaag
     2401 gtggtacggc ccctccggac ctttctgcac acagtgcaga ggaaccagat gctcacgacc
     2461 ccgacctcag ccccacgcag cgtcatgaag tcctttatta agcgcaacac tcccctgcgc
     2521 atggacccca agtgcagctt cgtcgagaag gagcggcagc gcctggagca gaaggagcag
     2581 ctgcagaggg aactggagga gaagaagaag aaggaagagc agcagcgtct ggctgagcgg
     2641 cagctgcagg aggagcaaga gaagaaagcc aaggaggcag caggggccag caaggccctg
     2701 aatgtgactg tggacgtgca gtctccagct tgtacctcat atcagatgac tccgcaaggg
     2761 cacagggccc ctcccaagat caacccagat aactacggga tggatctgaa tagcgacgac
     2821 tccaccgatg atgaggccca tccccggaag cccatcccca cctgggcccg aggcaccccg
     2881 ctcagccagg ctatcattca ccagtactac cacccaccga accttctgga gctctttgga
     2941 accattctcc cactggactt ggaggatatc ttcaagaaga gcaagccccg ctatcacaag
     3001 cgcaccagct ctgctgtctg gaactcaccg cccctgcagg gcgccagggt ccccagcagc
     3061 ctggcctaca gcctgaagaa gcacaagctt tgactcgagc atgcatctag agggccctat
     3121 tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg
     3181 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     3241 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     3301 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     3361 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     3421 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3481 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3541 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3601 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3661 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3721 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3781 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     3841 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     3901 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3961 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4021 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4081 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4141 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4201 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     4261 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4321 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4381 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4441 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4501 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4561 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4621 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4681 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4741 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4801 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4861 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4921 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4981 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5041 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5101 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5161 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5221 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     5281 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     5341 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     5401 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     5461 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     5521 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5581 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     5641 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5701 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5761 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5821 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5881 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5941 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6001 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6061 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6121 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6181 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6241 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6301 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6361 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6421 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6481 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6541 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6601 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6661 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6721 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6781 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6841 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6901 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6961 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7021 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7081 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7141 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7201 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7261 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7321 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7381 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7441 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7501 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     7561 ctgacgtc
//
