LOCUS       sequence-372333-        7672 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of CENP-C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7672)
  AUTHORS   Gascoigne KE, Takeuchi K, Suzuki A, Hori T, Fukagawa T, Cheeseman IM
  TITLE     Induced ectopic kinetochore assembly bypasses the requirement for 
            CENP-A nucleosomes.
  JOURNAL   Cell. 2011 Apr 29;145(3):410-22. doi: 10.1016/j.cell.2011.03.031.
  PUBMED    21529714
REFERENCE   2  (bases 1 to 7672)
  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: 
            "10.1016/j.cell.2011.03"; journalName: "Cell"; date: "2011-04-29- 
            29"; volume: "145"; issue: "3"; pages: "410-22"
COMMENT     Sequence Label: pKG139
FEATURES             Location/Qualifiers
     source          1..7672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        108..411
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        412..615
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     566..586
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      654..663
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             660..1367
                     /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(805..823)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1073..1092
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1413..1433
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1446..1490
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    4507..4628
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4544..4563)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4598..4617
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4635..5090)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4772..4793)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4984..5003
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5117..5221
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5223..5580
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5223..5414
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5246..5266)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5431..5566
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5615..6409
                     /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             5615..6409
                     /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(5669..5688)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6279..6298
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6597..6616)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6641..6688
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7017..7605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7506..7525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag
       61 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
      121 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      181 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      241 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
      301 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
      361 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      421 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      481 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      541 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      601 tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg gtcgccacca
      661 tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg
      721 tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc
      781 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct
      841 tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc
      901 ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg
      961 tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg
     1021 gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa
     1081 tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg
     1141 ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg
     1201 aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca
     1261 acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac
     1321 gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtcc ggaggggatc
     1381 ggtggagctc caccggtggc ggccgctcta gagagaatct ttattttcag ggcgccgcca
     1441 aattcaaaga aaccgctgct gctaaattcg aacgccagca catggacagc ggaggtggag
     1501 gtactagtct cgagatggct gcgtccggtc tggatcatct caaaaatggc tacagaagaa
     1561 gattttgtcg accttccagg gcacgtgaca ttaacacaga gcaaggccag aatgttctgg
     1621 aaatcttaca agactgtttt gaagaaaaaa gtcttgccaa tgattttagt acaaattcta
     1681 caaaatcagt gcctaattca acacgcaaaa taaaagacac ttgtattcag tcaccaagca
     1741 aagagtgcca gaaatcacat ccaaagtcag ttccagtttc ttcaaagaag aaagaagcct
     1801 ctctacagtt tgttgtagaa ccaagtgaag ccacaaacag atcagttcag gcccatgaag
     1861 ttcatcagaa aattctggca actgatgtta gttccaaaaa tacacctgac tcgaaaaaaa
     1921 tatcaagtag aaacataaat gatcatcaca gtgaagctga tgaagaattt tacttatccg
     1981 ttggctcacc ttctgttctt ttggatgcaa aaacatctgt atcacaaaat gttattccat
     2041 ctagtgccca aaagagagag acttacactt ttgaaaattc agtaaatatg ctgccttcaa
     2101 gtacagaggt ttcagttaaa accaaaaaaa ggttaaactt tgatgataaa gttatgttaa
     2161 agaaaataga aatagataat aaagtatcag atgaagagga taaaacatca gaaggacaag
     2221 aaagaaaacc atcaggatca tctcagaata gaatacgaga ttcagaatat gaaattcaac
     2281 gacaagctaa aaaaagtttt tcaacattgt ttttagaaac agtaaaacga aaaagtgaat
     2341 ccagtcccat tgttaggcat gcggcaactg ctccacctca ttcgtgtcct cccgatgata
     2401 cgaagttgat agaggatgaa tttataattg atgagtcgga tcaaagtttt gccagtagat
     2461 cttggattac aataccaaga aaggcagggt ctctgaaaca acgcacaata tccccggctg
     2521 agagcactgc actctttcaa ggtagaaagt caagagaaaa gcatcataat atattaccta
     2581 agactttggc aaatgacaaa cattcccata aacctcaccc agtagagaca tctcagccct
     2641 ctgataaaac agtactggat acaagttatg ctttgatagg tgaaacagta aataattata
     2701 gatctacaaa atatgaaatg tattccaaga atgcagaaaa accatctaga agcaaaagga
     2761 ctataaaaca aaaacagaga agaaaattca tggctaaacc agctgaagaa cagcttgatg
     2821 tgggacagtc taaagatgaa aacatacata catcacatat tacccaagac gaatttcaaa
     2881 gaaattcaga cagaaatatg gaagagcatg aagagatggg aaatgattgt gtttccaaaa
     2941 aacagatgcc acctgtggga agcaagaaaa gtagcactag aaaagataag gaagaatcta
     3001 aaaagaagcg cttttccagt gagtccaaga acaaacttgt acctgaagaa gtgacttcaa
     3061 ctgtcacgaa aagtcgaaga atttccaggc gtccatctga ttggtgggtg gtaaaatcag
     3121 aggagagtcc tgtttatagc aattcttcag taagaaatga attaccaatg catcacaata
     3181 gtagccgaaa atctactaag aaaacaaatc agtcatctaa ggatattagg aaaaaaacta
     3241 ttccacttaa aaggcagaag acagcaacta aaggcaacca aagagtacag aagtttttaa
     3301 atgctgaagg ttctggaggt atcgttggtc atgatgaaat ttccagatgt tcactgagtg
     3361 agccattgga aagtgatgag gcagacttgg ctaagaagaa aaatcttgat tgttctagat
     3421 ctacaagaag ctcaaagaat gaagataaca ttatgactgc acagaatgtt cccctaaagc
     3481 ctcagaccag tggatataca tgtaatatac caacagagtc aaacttggat tctggagagc
     3541 ataagacttc agttttagag gaaagtggac cttccaggct caataataat tatttaatgt
     3601 ctggaaagaa tgatgtggat gatgaggaag ttcatggaag ttcagatgac tcaaaacaat
     3661 ctaaagtgat accaaagaac agaatccatc acaaactagt attgccctcc aacacaccaa
     3721 atgttcgcag gaccaagaga acacgtttga aacctttgga gtactggcga ggagagcgaa
     3781 tagattatca aggaaggcca tcaggaggat tcgtgattag tggagtacta tctccagaca
     3841 caatatcgtc taaaaggaag gcaaaagaaa atattggaaa agtcaacaaa aaatctaata
     3901 agaaaaggat ctgtcttgat aacgatgaaa gaaagactaa cttaatggta aatctaggta
     3961 tacctcttgg agatcctttg cagccaacga gggtaaagga cccagaaaca agagagatta
     4021 ttctcatgga tcttgtaagg ccacaagata catatcaatt ttttgttaag catggtgagt
     4081 tgaaggtata caagacattg gatacaccct ttttttctac tgggaaattg atattaggac
     4141 cacaagaaga aaagggaaag cagcatgttg gccaggatat attggttttt tatgttaact
     4201 ttggtgacct tttgtgtact ttacatgaaa caccttatat attaagtact ggggattcgt
     4261 tctatgttcc ttcaggtaac tattataaca tcaaaaatct ccggaatgag gaaagtgttc
     4321 ttctttttac tcagataaaa agatgagaat tctgcagtcg acggtaccgc gggcccggga
     4381 tccaccggat ctagataact gatcataatc agccatacca catttgtaga ggttttactt
     4441 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     4501 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4561 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4621 gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     4681 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     4741 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     4801 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     4861 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     4921 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     4981 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     5041 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     5101 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     5161 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     5221 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     5281 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     5341 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5401 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     5461 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     5521 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     5581 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     5641 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     5701 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     5761 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     5821 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     5881 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     5941 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     6001 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     6061 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     6121 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     6181 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     6241 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     6301 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     6361 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     6421 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     6481 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     6541 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     6601 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     6661 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     6721 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     6781 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     6841 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     6901 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6961 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     7021 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     7081 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     7141 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     7201 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     7261 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     7321 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     7381 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     7441 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7501 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7561 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     7621 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tc
//
