LOCUS       sequence-208158-        7517 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 7517)
  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 7517)
  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..7517
                     /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    4305..4426
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4342..4361)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4396..4415
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4433..4888)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4570..4591)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4782..4801
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4915..5019
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5021..5378
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5021..5212
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5044..5064)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5229..5364
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5413..6207
                     /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             5413..6207
                     /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(5467..5486)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6077..6096
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6395..6414)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6439..6486
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6815..7403
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7304..7323
                     /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 agcaggggcc agcaaggccc tgaatgtgac tgtggacgtg
     3061 cagtctccag cttgtacctc atatcagatg 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 aaagcggccg cgactctaga
     4201 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     4261 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     4321 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4381 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta
     4441 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     4501 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     4561 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     4621 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     4681 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     4741 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     4801 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     4861 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4921 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4981 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     5041 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     5101 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     5161 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     5221 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     5281 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     5341 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     5401 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     5461 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     5521 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     5581 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     5641 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     5701 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     5761 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     5821 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5881 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5941 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     6001 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     6061 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     6121 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     6181 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     6241 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     6301 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     6361 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     6421 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     6481 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     6541 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     6601 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     6661 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6721 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6781 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6841 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6901 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6961 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     7021 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     7081 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     7141 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     7201 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     7261 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     7321 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     7381 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     7441 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     7501 gtattaccgc catgcat
//
