LOCUS       sequence-206165-        9684 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express tagged Cdc6 mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9684)
  AUTHORS   Matson JP, Dumitru R, Coryell P, Baxley RM, Chen W, Twaroski K, 
            Webber BR, Tolar J, Bielinsky AK, Purvis JE, Cook JG
  TITLE     Rapid DNA replication origin licensing protects stem cell 
            pluripotency.
  JOURNAL   Elife. 2017 Nov 17;6. doi: 10.7554/eLife.30473.
  PUBMED    29148972
REFERENCE   2  (bases 1 to 9684)
  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"; journalName: "Elife."; 
            date: "2017-11-17"; volume: "6. doi"; pages: " 10.7554/eLife.30473"
COMMENT     Sequence Label: CLXSN 5myc-CDC6-mut
FEATURES             Location/Qualifiers
     source          1..9684
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        7..385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..588
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     539..559
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             589..764
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    827..1026
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1227..1643
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1573..1595
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1611..1627
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1692..1781
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     CDS             1722..1811
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     CDS             1752..1841
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     primer_bind     complement(3550..3570)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3555..3884
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3735..3870
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3944..4738
                     /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(3998..4017)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4608..4627
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             4798..5391
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     intron          5738..5803
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             5932..5952
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    6224..6345
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6261..6280)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6315..6334
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7390..7407)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7561..8149)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7641..7660)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8618..8994)
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     primer_bind     8941..8960
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9179..9283)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9351..9369
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     9529..9548
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttccatag taacgccaat
      181 agggactttc cattgacgtc aatgggtgga ctatttacgg taaactgccc acttggcagt
      241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      361 cgtattagtc atcgctatta ccatgtgatg cggttttggc agtacatcaa tgggcgtgga
      421 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      481 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      541 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctca ataaaagagc
      601 ccacaacccc tcactcggcg cgccagtctt ccgatagact gcgtcgcccg ggtacccgta
      661 ttcccaataa agcctcttgc tgtttgcatc cgaatcgtgg tctcgctgtt ccttgggagg
      721 gtctcctctg agtgattgac tacccacgac gggggtcttt catttggggg ctcgtccggg
      781 atttggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc tggccagcaa
      841 cttatctgtg tctgtccgat tgtctagtgt ctatgtttga tgttatgcgc ctgcgtctgt
      901 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      961 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
     1021 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
     1081 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
     1141 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
     1201 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1261 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1321 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1381 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1441 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1501 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1561 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1621 ccctcactcc ttctctaggc gccggaattc gttaacagga accaattcag tcgactggat
     1681 ccggtaccat ggaacaaaag ctgatttcgg aagaggatct ggaacaaaag ctgatttcgg
     1741 aagaggatct ggaacaaaag ctgatttcgg aagaggatct ggaacaaaag ctgatttcgg
     1801 aagaggatct ggaacaaaag ctgatttcgg aagaggatct ggggatcatg cctcaaaccc
     1861 gatcccaggc acaggctaca atcagttttc caaaaaggaa gctgtctcgg gcattgaaca
     1921 aagctaaaaa ctccagtgat gccaaactag aaccaacaaa tgtccaaacc gtaacctgtt
     1981 ctcctcgtgt aaaagccctg cctctcagcc ccgccaaacg tgccggcgat gacaacctat
     2041 gcaacactcc ccatttacct ccttgttctc caccaaagca aggcaaggcg gccgcaggtc
     2101 cccctcactc acatacactt aagggacgaa gattggtatt tgacaatcag ctgacaatta
     2161 agtctcctag caaaagagaa ctagccaaag ttcaccaaaa caaaatactt tcttcagtta
     2221 gaaaaagtca agagatcaca acaaattctg agcagagatg tccactgaag aaagaatctg
     2281 catgtgtgag actattcaag caagaaggca cttgctacca gcaagcaaag ctggtcctga
     2341 acacagctgt cccagatcgg ctgcctgcca gggaaaggga gatggatgtc atcaggaatt
     2401 tcttgaggga acacatctgt gggaaaaaag ctggaagcct ttacctttct ggtgctcctg
     2461 gaactggaaa aactgcctgc ttaagccgga ttctgcaaga cctcaagaag gaactgaaag
     2521 gctttaaaac tatcatgctg aattgcatgt ccttgaggac tgcccaggct gtattcccag
     2581 ctattgctca ggagatttgt caggaagagg tatccaggcc agctgggaag gacatgatga
     2641 ggaaattgga aaaacatatg actgcagaga agggccccat gattgtgttg gtattggacg
     2701 agatggatca actggacagc aaaggccagg atgtattgta cacgctattt gaatggccat
     2761 ggctaagcaa ttctcacttg gtgctgattg gtattgctaa taccctggat ctcacagata
     2821 gaattctacc taggcttcaa gctagagaaa aatgtaagcc acagctgttg aacttcccac
     2881 cttataccag aaatcagata gtcactattt tgcaagatcg acttaatcag gtatctagag
     2941 atcaggttct ggacaatgct gcagttcaat tctgtgcccg caaagtctct gctgtttcag
     3001 gagatgttcg caaagcactg gatgtttgca ggagagctat tgaaattgta gagtcagatg
     3061 tcaaaagcca gactattctc aaaccactgt ctgaatgtaa atcaccttct gagcctctga
     3121 ttcccaagag ggttggtctt attcacatat cccaagtcat ctcagaagtt gatggtaaca
     3181 ggatgacctt gagccaagaa ggagcacaag attccttccc tcttcagcag aagatcttgg
     3241 tttgctcttt gatgctcttg atcaggcagt tgaaaatcaa agaggtcact ctggggaagt
     3301 tatatgaagc ctacagtaaa gtctgtcgca aacagcaggt ggcggctgtg gaccagtcag
     3361 agtgtttgtc actttcaggg ctcttggaag ccaggggcat tttaggatta aagagaaaca
     3421 aggaaacccg tttgacaaag gtgtttttca agattgaaga gaaagaaata gaacatgctc
     3481 tgaaagataa agctttaatt ggaaatatct tagctactgg attgccttaa gcgtcgagga
     3541 tccggctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3601 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3661 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3721 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3781 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     3841 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt gggctgcagg
     3901 tcgaggcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     3961 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4021 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4081 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     4141 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4201 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4261 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4321 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4381 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     4441 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     4501 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4561 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     4621 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     4681 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     4741 gggactctgg ggttcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     4801 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     4861 gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagc
     4921 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     4981 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     5041 cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga
     5101 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
     5161 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
     5221 aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt
     5281 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt
     5341 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcat ttgggggctc
     5401 gtccgggatc gggagacccc tgcccaggga ccaccgaccc accaccggga ggtaagctgg
     5461 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     5521 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     5581 gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta
     5641 tacctagcta ggtagctaga ggatctttgt gaaggaacct tacttctgtg gtgtgacata
     5701 attggacaaa ctacctacag agatttaaag ctctaaggta aatataaaat ttttaagtgt
     5761 ataatgtgtt aaactactga ttctaattgt ttgtgtattt tagattccaa cctatggaac
     5821 tgatgaatgg gagcagtggt ccaatgcctt taatgaggaa aacctgtttt gctcagaaga
     5881 aatgcctcta gtgatgatga ggctactgct gactctcaac attctactcc tccaaaaaag
     5941 aagagaaagg tagaagaccc caaggacttt ccttcagaat tgctaagttt tttgagtcat
     6001 gctgtgttta gtaatagaac tcttgcttgc tttgctattt acaccacaaa ggaaaaagct
     6061 gcactgctat acaagaaaat tatggaaaaa tatttgatgt atagtgcctt gactagagat
     6121 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     6181 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     6241 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     6301 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgatcccg
     6361 ccatggtatc aacgccatat ttctatttac agtagggacc tcttcgttgt gtaggtaccg
     6421 ctgtattcct agggaaatag tagaggcacc ttgaactgtc tgcatcagcc atatagcccc
     6481 cgctgttcga cttacaaaca caggcacagt actgacaaac ccatacacct cctctgaaat
     6541 acccatagtt gctagggctg tctccgaact cattacaccc tccaaagtca gagctgtaat
     6601 ttcgccatca agggcagcga gggcttctcc agataaaata gcttctgccg agagtcccgt
     6661 aagggtagac acttcagcta atccctcgat gaggtctact agaatagtca gtgcggctcc
     6721 cattttgaaa attcacttac ttgatcagct tcagaagatg gcggagggcc tccaacacag
     6781 taattttcct cccgactctt aaaatagaaa atgtcaagtc agttaagcag gaagtggact
     6841 aactgacgca gctggccgtg cgacatcctc ttttaattag ttgctaggca acgccctcca
     6901 gagggcgtgt ggttttgcaa gaggaagcaa aagcctctcc acccaggcct agaatgtttc
     6961 cacccaatca ttactatgac aacagctgtt ttttttagta ttaagcagag gccggggacc
     7021 cctggcccgc ttactctgga gaaaaaaaac attgtagagg cttccagagg caacttgtca
     7081 aaacaggact gcttctattt ctgtcacact gtctggccct gtcacaaggt ccagcacctc
     7141 cataccccct ttaataagca gtttgggaac gggtgcgggt cttactccgc ccatccgccc
     7201 ctaactccgc ccagttccgc ccattctccg ccccatgctg actaattttt tttatttatg
     7261 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     7321 gaggctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     7381 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     7441 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     7501 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     7561 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     7621 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     7681 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     7741 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     7801 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7861 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7921 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7981 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     8041 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     8101 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     8161 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     8221 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     8281 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     8341 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     8401 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     8461 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     8521 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     8581 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctgcagg
     8641 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     8701 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     8761 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     8821 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg aactcaacca
     8881 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     8941 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     9001 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     9061 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     9121 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     9181 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     9241 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     9301 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     9361 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     9421 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     9481 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     9541 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     9601 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     9661 tgcgggcctc ttcgctatta cgcc
//
