LOCUS       sequence-205258-        7515 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of vsv-INCENP wt-mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7515)
  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 7515)
  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: mCherry-Incenp wt
FEATURES             Location/Qualifiers
     source          1..7515
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             643..675
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     CDS             3472..4182
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3617..3635)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3885..3904
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4303..4424
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4340..4359)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4394..4413
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4431..4886)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4568..4589)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4780..4799
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4913..5017
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5019..5376
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5019..5210
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5042..5062)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5227..5362
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5411..6205
                     /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             5411..6205
                     /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(5465..5484)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6075..6094
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6393..6412)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6437..6484
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6813..7401
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7302..7321
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttggt accgccacca tgtacaccga catcgagatg
      661 aaccggttgg gcaaggggac gacggcccca gggcccattc acctgctgga gctctgtgac
      721 cagaagctca tggagtttct ctgcaacatg gataataagg acttggtgtg gcttgaggag
      781 atccaagagg aggccgagcg catgttcacc agagaattca gcaaagagcc agagctgatg
      841 cccaaaacac cttctcagaa gaaccgacgg aagaagagac ggatttctta tgttcaggat
      901 gaaaacagag atcccatcag gagaaggtta tcccgcagaa agtctcggag cagccagctg
      961 agctcccgac gcctccgcag caaggacagt gtagagaagc tggctacagt ggtcggggag
     1021 aacggctccg ttctgcggcg tgtgacccgt gctgcggctg cagctgccgc ggctaccatg
     1081 gcattggctg caccttcttc acccacccct gagtctccca cgatgctgac taagaagccc
     1141 gaggataacc acacccagtg ccagctggtg cctgtggtgg agatcggcat cagtgagcgc
     1201 cagaatgctg agcagcatgt cacccagctc atgtccaccg agcctctgcc ccgcactctg
     1261 tccccgactc cagcttcagc cacagctcca acctcccagg gcatcccgac atcagatgag
     1321 gaatcaacac ctaagaagtc gaaggccagg atactggagt ccatcacagt gagctccctg
     1381 atggctacac cccaggaccc caagggtcaa ggggtcggga cggggcggtc tgcgtctaag
     1441 ctcaggattg cgcaggtctc ccctggccca cgggactcgc cagcctttcc agattctcca
     1501 tggcgggagc gggtgctggc tcccatcctg ccggataact tctccacgcc cacgggctct
     1561 cgcacggact ctcaatcggt gcggcacagc ccgatcgccc cgtcttcccc gagtccccga
     1621 gtcttagccc agaagtactc tctggtggcc aaacaggaaa gtgttgtccg cagggcgagc
     1681 agaaggcttg ccaagaagac tgccgaagag ccagctgcct ctggccgcat catctgtcac
     1741 agttacctgg agaggctcct gaatgttgag gtgccccaga aagttggttc tgagcagaag
     1801 gaaccccccg aggaggctga gcctgtggcg gcagctgagc cagaggtccc tgagaacaac
     1861 ggaaataact cgtggcccca caatgacaca gagatcgcca acagcacacc caacccgaag
     1921 cctgcagcca gcagcccgga aacaccctct gcagggcagc aagaggccaa gacggaccaa
     1981 gcagatggac ccagagagcc accgcagagt gccaggagga agcgcagcta caagcaggcc
     2041 gtgagtgagc tggacgagga gcagcacctg gaggatgagg agctgcagcc ccccaggagc
     2101 aagacccctt cctcaccctg cccagccagc aaggtggtac ggcccctccg gacctttctg
     2161 cacacagtgc agaggaacca gatgctcacg accccgacct cagccccacg cagcgtcatg
     2221 aagtccttta ttaagcgcaa cactcccctg cgcatggacc ccaagtgcag cttcgtcgag
     2281 aaggagcggc agcgcctgga gaatctgcgg cggaaggagg aggccgagca gctgcgcagg
     2341 cagaaggtgg aggaggacaa gcggcggcgg ctggaggagg tgaagctgaa gcgtgaggaa
     2401 cgcctccgca aggtgctgca ggcccgcgag cgggtggagc agatgaagga ggagaagaag
     2461 aagcagattg agcagaagtt tgctcagatc gacgagaaga ctgagaaggc caaggaggag
     2521 cggctggcag aggagaaggc caagaaaaag gcggcggcca agaagatgga ggaggtggaa
     2581 gcacgcagga agcaggaaga ggatgcacgt aggctcaggt ggctgcagca ggaggaggaa
     2641 gagcggcggc accaagagct gctgcagaag aagaaggaag aggagcagga gcggctgcgg
     2701 aaggcggccg aggctaagcg gctggcagag cagcgggagc aggagcggcg ggagcaggag
     2761 cggcgcgagc aggagcggcg cgagcaggag cggcgggagc aggagcggcg cgagcaggag
     2821 cgacagctgg cagagcagga gcgtcggcgg gagcaggagc ggctccaggc cgagagggag
     2881 ctgcaggagc gggagaaggc cctgcggctg cagaaggagc agctgcagag ggaactggag
     2941 gagaagaaga agaaggaaga gcagcagcgt ctggctgagc ggcagctgca ggaggagcaa
     3001 gagaagaaag ccaaggaggc agtaggggcc agcaaggccc tgaatgtgac tgtggacgtg
     3061 cagtctccag cttgtacctc atatcaaatg actccgcaag ggcacagggc ccctcccaag
     3121 atcaacccag ataactacgg gatggatctg aatagcgacg actccaccga tgatgaggcc
     3181 catccccgga agcccatccc cacctgggcc cgaggcaccc cgctcagcca ggctatcatt
     3241 caccagtact accacccacc gaaccttctg gagctctttg gaaccattct cccactggac
     3301 ttggaggata tcttcaagaa gagcaagccc cgctatcaca agcgcaccag ctctgctgtc
     3361 tggaactcac cgcccctgca gggcgccagg gtccccagca gcctggccta cagcctgaag
     3421 aagcacaagc ttcgaattct gcagtcgacg gtaccgcggg cccgggatcc aatggtgagc
     3481 aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg
     3541 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     3601 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg
     3661 gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac
     3721 atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
     3781 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc
     3841 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag
     3901 aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag
     3961 ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag
     4021 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag
     4081 ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag
     4141 ggccgccact ccaccggcgg catggacgag ctgtacaagt agcggccgcg actctagatc
     4201 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     4261 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     4321 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     4381 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag
     4441 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     4501 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     4561 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     4621 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     4681 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     4741 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     4801 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     4861 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     4921 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4981 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     5041 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5101 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     5161 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     5221 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5281 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     5341 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     5401 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     5461 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     5521 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     5581 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     5641 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     5701 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     5761 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     5821 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     5881 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     5941 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     6001 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     6061 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     6121 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     6181 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     6241 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     6301 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     6361 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     6421 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     6481 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     6541 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     6601 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     6661 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6721 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6781 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6841 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6901 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     6961 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7021 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7081 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7141 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7201 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7261 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7321 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7381 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7441 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     7501 attaccgcca tgcat
//
