LOCUS       sequence-208716-        7669 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of vsv-dCEN-INCENP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7669)
  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 7669)
  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-dCEN-Incenp
FEATURES             Location/Qualifiers
     source          1..7669
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        6..385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..589
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     540..560
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     634..653
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        634..652
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             687..719
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     promoter        complement(3374..3392)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3375..3392)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3412..3429)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3418..3642
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3774..4362)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3854..3873)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4691..4738
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4763..4782
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4970..5764)
                     /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(4970..5764)
                     /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(5081..5100)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5691..5710
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5799..6156)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5813..5948
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5965..6156
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6113..6133
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(6216..7007,7008..7076))
                     /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     6839..6858
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7077..7181)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7208..7663
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7295..7314)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7505..7526
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
       61 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      121 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      181 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      361 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      421 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      481 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      541 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac
      601 tagagaaccc actgcttact ggcttatcga aattaatacg actcactata gggagaccca
      661 agcttggtac cggatccgcc accatgtaca ccgacatcga gatgaaccgg ttgggcaagg
      721 aattcagcaa agagccagag ctgatgccca aaacaccttc tcagaagaac cgacggaaga
      781 agagacggat ttcttatgtt caggatgaaa acagagatcc catcaggaga aggttatccc
      841 gcagaaagtc tcggagcagc cagctgagct cccgacgcct ccgcagcaag gacagtgtag
      901 agaagctggc tacagtggtc ggggagaacg gctccgttct gcggcgtgtg acccgtgctg
      961 cggctgcagc tgccgcggct accatggcat tggctgcacc ttcttcaccc acccctgagt
     1021 ctcccacgat gctgactaag aagcccgagg ataaccacac ccagtgccag ctggtgcctg
     1081 tggtggagat cggcatcagt gagcgccaga atgctgagca gcatgtcacc cagctcatgt
     1141 ccaccgagcc tctgccccgc actctgtccc cgactccagc ttcagccaca gctccaacct
     1201 cccagggcat cccgacatca gatgaggaat caacacctaa gaagtcgaag gccaggatac
     1261 tggagtccat cacagtgagc tccctgatgg ctacacccca ggaccccaag ggtcaagggg
     1321 tcgggacggg gcggtctgcg tctaagctca ggattgcgca ggtctcccct ggcccacggg
     1381 actcgccagc ctttccagat tctccatggc gggagcgggt gctggctccc atcctgccgg
     1441 ataacttctc cacgcccacg ggctctcgca cggactctca atcggtgcgg cacagcccga
     1501 tcgccccgtc ttccccgagt ccccgagtct tagcccagaa gtactctctg gtggccaaac
     1561 aggaaagtgt tgtccgcagg gcgagcagaa ggcttgccaa gaagactgcc gaagagccag
     1621 ctgcctctgg ccgcatcatc tgtcacagtt acctggagag gctcctgaat gttgaggtgc
     1681 cccagaaagt tggttctgag cagaaggaac cccccgagga ggctgagcct gtggcggcag
     1741 ctgagccaga ggtccctgag aacaacggaa ataactcgtg gccccacaat gacacagaga
     1801 tcgccaacag cacacccaac ccgaagcctg cagccagcag cccggaaaca ccctctgcag
     1861 ggcagcaaga ggccaagacg gaccaagcag atggacccag agagccaccg cagagtgcca
     1921 ggaggaagcg cagctacaag caggccgtga gtgagctgga cgaggagcag cacctggagg
     1981 atgaggagct gcagcccccc aggagcaaga ccccttcctc accctgccca gccagcaagg
     2041 tggtacggcc cctccggacc tttctgcaca cagtgcagag gaaccagatg ctcacgaccc
     2101 cgacctcagc cccacgcagc gtcatgaagt cctttattaa gcgcaacact cccctgcgca
     2161 tggaccccaa gtgcagcttc gtcgagaagg agcggcagcg cctggagaat ctgcggcgga
     2221 aggaggaggc cgagcagctg cgcaggcaga aggtggagga ggacaagcgg cggcggctgg
     2281 aggaggtgaa gctgaagcgt gaggaacgcc tccgcaaggt gctgcaggcc cgcgagcggg
     2341 tggagcagat gaaggaggag aagaagaagc agattgagca gaagtttgct cagatcgacg
     2401 agaagactga gaaggccaag gaggagcggc tggcagagga gaaggccaag aaaaaggcgg
     2461 cggccaagaa gatggaggag gtggaagcac gcaggaagca ggaagaggat gcacgtaggc
     2521 tcaggtggct gcagcaggag gaggaagagc ggcggcacca agagctgctg cagaagaaga
     2581 aggaagagga gcaggagcgg ctgcggaagg cggccgaggc taagcggctg gcagagcagc
     2641 gggagcagga gcggcgggag caggagcggc gcgagcagga gcggcgcgag caggagcggc
     2701 gggagcagga gcggcgcgag caggagcgac agctggcaga gcaggagcgt cggcgggagc
     2761 aggagcggct ccaggccgag agggagctgc aggagcggga gaaggccctg cggctgcaga
     2821 aggagcagct gcagagggaa ctggaggaga agaagaagaa ggaagagcag cagcgtctgg
     2881 ctgagcggca gctgcaggag gagcaagaga agaaagccaa ggaggcagca ggggccagca
     2941 aggccctgaa tgtgactgtg gacgtgcagt ctccagcttg tacctcatat cagatgactc
     3001 cgcaagggca cagggcccct cccaagatca acccagataa ctacgggatg gatctgaata
     3061 gcgacgactc caccgatgat gaggcccatc cccggaagcc catccccacc tgggcccgag
     3121 gcaccccgct cagccaggct atcattcacc agtactacca cccaccgaac cttctggagc
     3181 tctttggaac cattctccca ctggacttgg aggatatctt caagaagagc aagccccgct
     3241 atcacaagcg caccagctct gctgtctgga actcaccgcc cctgcagggc gccagggtcc
     3301 ccagcagcct ggcctacagc ctgaagaagc acaagctttg actcgagtag atgactagtc
     3361 tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg
     3421 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     3481 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     3541 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     3601 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     3661 ccagtggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3721 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3781 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3841 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3901 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3961 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4021 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4081 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4141 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4201 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4261 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4321 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4381 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4441 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4501 taaagtatat atgagtaacc tgaggctatg gcagggcctg ccgccccgac gttggctgcg
     4561 agccctgggc cttcacccga acttgggggg tggggtgggg aaaaggaaga aacgcgggcg
     4621 tattggcccc aatggggtct cggtggggta tcgacagagt gccagccctg ggaccgaacc
     4681 ccgcgtttat gaacaaacga cccaacaccg tgcgttttat tctgtctttt tattgccgtc
     4741 atagcgcggg ttccttccgg tattgtctcc ttccgtgttt cagttagcct ccccctaggg
     4801 tgggcgaaga actccagcat gagatccccg cgctggagga tcatccagcc ggcgtcccgg
     4861 aaaacgattc cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa atctcgtgat
     4921 ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct cagaagaact
     4981 cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca
     5041 cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg
     5101 ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc
     5161 ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct
     5221 cgccgtcggg catgctcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat
     5281 gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct
     5341 cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc
     5401 gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga
     5461 gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt
     5521 cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt
     5581 cttgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct
     5641 gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat
     5701 agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa
     5761 tcatgcgaaa cgatcctcat cctgtctctt gatcgatctt tgcaaaagcc taggcctcca
     5821 aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca
     5881 taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg
     5941 ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt
     6001 tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg
     6061 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac
     6121 tgacacacat tccacagctg gttctttccg cctcaggact cttccttttt caataaatca
     6181 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6241 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6301 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6361 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6421 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6481 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6541 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6601 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6661 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6721 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6781 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6841 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6901 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6961 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7021 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7081 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7141 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7201 ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     7261 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     7321 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     7381 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     7441 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     7501 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     7561 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     7621 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttacgcgc
//
