LOCUS Exported 9473 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION express tagged Cdc6 WT. ACCESSION . VERSION . KEYWORDS CLXSN 5myc-CDC6-WT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9473) 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 9473) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9473 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 7..386 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 387..590 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 541..561 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 591..766 /label=5' LTR (truncated) /note="truncated long terminal repeat from Moloney murine sarcoma virus" misc_feature 829..1028 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 1229..1645 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1575..1597 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1613..1629 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" primer_bind 1707..1726 /label=T7 /note="T7 promoter, forward primer" promoter 1707..1725 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 1793..1822 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 1826..1855 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 1865..1894 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 1898..1927 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" promoter complement(3671..3689) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(3672..3689) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(3711..3731) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3716..4045 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3896..4031 /label=SV40 ori /note="SV40 origin of replication" CDS 4105..4899 /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(4159..4178) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4769..4788 /label=Neo-F /note="Neomycin resistance gene, forward primer" LTR 4959..5552 /note="long terminal repeat from Moloney murine leukemia virus" intron 5897..5962 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 6092..6112 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 6384..6518 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6421..6440) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6475..6494 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 6976..7110 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(7175..7192) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7346..7934) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7426..7445) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8105..8965) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 8728..8747 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8966..9070) /gene="bla" /label=AmpR promoter primer_bind 9138..9156 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind 9316..9335 /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 tgttcccata gtaacgccaa 181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag 241 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc 301 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct 361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg 421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt 481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga 541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct caataaaaga 601 gcccacaacc cctcactcgg cgcgccagtc ttccgataga ctgcgtcgcc cgggtacccg 661 tattcccaat aaagcctctt gctgtttgca tccgaatcgt ggtctcgctg ttccttggga 721 gggtctcctc tgagtgattg actacccacg acgggggtct ttcatttggg ggctcgtccg 781 ggatttggag acccctgccc agggaccacc gacccaccac cgggaggtaa gctggccagc 841 aacttatctg tgtctgtccg attgtctagt gtctatgttt gatgttatgc gcctgcgtct 901 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg 961 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc 1021 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg 1081 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg 1141 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac 1201 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac 1261 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa 1321 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg 1381 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg 1441 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga 1501 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc 1561 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc 1621 agccctcact ccttctctag gcgccggaat tcgttaactc gactctggct aactagagaa 1681 cccactgctt actggcttat cgaaattaat acgactcact atagggagac ccaagcttgg 1741 taccaccatg gaacagaagc taatctctga ggaagttttg ctggggatct tggagcagaa 1801 attgataagc gaggaagacc ttggagaaca aaagctgatt tcggaagagg atctggggat 1861 cttggagcag aaattgataa gcgaggaaga ccttggagaa caaaagctga tttcggaaga 1921 ggatctgggg atccgaatgc ctcaaacccg atcccaggca caggctacaa tcagttttcc 1981 aaaaaggaag ctgtctcggg cattgaacaa agctaaaaac tccagtgatg ccaaactaga 2041 accaacaaat gtccaaaccg taacctgttc tcctcgtgta aaagccctgc ctctcagccc 2101 caggaaacgt ctgggcgatg acaacctatg caacactccc catttacctc cttgttctcc 2161 accaaagcaa ggcaagaaag agaatggtcc ccctcactca catacactta agggacgaag 2221 attggtattt gacaatcagc tgacaattaa gtctcctagc aaaagagaac tagccaaagt 2281 tcaccaaaac aaaatacttt cttcagttag aaaaagtcaa gagatcacaa caaattctga 2341 gcagagatgt ccactgaaga aagaatctgc atgtgtgaga ctattcaagc aagaaggcac 2401 ttgctaccag caagcaaagc tggtcctgaa cacagctgtc ccagatcggc tgcctgccag 2461 ggaaagggag atggatgtca tcaggaattt cttgagggaa cacatctgtg ggaaaaaagc 2521 tggaagcctt tacctttctg gtgctcctgg aactggaaaa actgcctgct taagccggat 2581 tctgcaagac ctcaagaagg aactgaaagg ctttaaaact atcatgctga attgcatgtc 2641 cttgaggact gcccaggctg tattcccagc tattgctcag gagatttgtc aggaagaggt 2701 atccaggcca gctgggaagg acatgatgag gaaattggaa aaacatatga ctgcagagaa 2761 gggccccatg attgtgttgg tattggacga gatggatcaa ctggacagca aaggccagga 2821 tgtattgtac acgctatttg aatggccatg gctaagcaat tctcacttgg tgctgattgg 2881 tattgctaat accctggatc tcacagatag aattctacct aggcttcaag ctagagaaaa 2941 atgtaagcca cagctgttga acttcccacc ttataccaga aatcagatag tcactatttt 3001 gcaagatcga cttaatcagg tatctagaga tcaggttctg gacaatgctg cagttcaatt 3061 ctgtgcccgc aaagtctctg ctgtttcagg agatgttcgc aaagcactgg atgtttgcag 3121 gagagctatt gaaattgtag agtcagatgt caaaagccag actattctca aaccactgtc 3181 tgaatgtaaa tcaccttctg agcctctgat tcccaagagg gttggtctta ttcacatatc 3241 ccaagtcatc tcagaagttg atggtaacag gatgaccctg agccaagaag gagcacaaga 3301 ttccttccct cttcagcaga agatcttggt ttgctctttg atgctcttga tcaggcagtt 3361 gaaaatcaaa gaggtcactc tggggaagtt atatgaagcc tacagtaaag tctgtcgcaa 3421 acagcaggtg gcggctgtgg accagtcaga gtgtttgtca ctttcagggc tcttggaagc 3481 caggggcatt ttaggattaa agagaaacaa ggaaacccgt ttgacaaagg tgtttttcaa 3541 gattgaagag aaagaaatag aacatgctct gaaagataaa gctttaattg gaaatatctt 3601 agctactgga ttgccttaag gatcgggtcg actcgagcgg ccgctcgagc atgcatctag 3661 agggccctat tctatagtgt cacctaaatg ctagtcgagg atccggctgt ggaatgtgtg 3721 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca 3781 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 3841 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc 3901 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat 3961 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt 4021 ttttggaggc ctaggctttt gcaaaaagct tgggctgcag gtcgaggcgg atctgatcaa 4081 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg 4141 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct 4201 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac 4261 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg 4321 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg 4381 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa 4441 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca 4501 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt 4561 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc 4621 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc 4681 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg 4741 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt 4801 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag 4861 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgata 4921 aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg 4981 tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaaatac ataactgaga 5041 atagagaagt tcagatcaag gtcaggaaca gatggaacag ctgaatatgg gccaaacagg 5101 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg aacagctgaa 5161 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca 5221 gatggtcccc agatgcggtc cagccctcag cagtttctag agaaccatca gatgtttcca 5281 gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct 5341 tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc 5401 actcggggcg ccagtcctcc gattgactga gtcgcccggg tacccgtgta tccaataaac 5461 cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt 5521 gattgactac ccgtcagcgg gggtctttca tttgggggct cgtccgggat cgggagaccc 5581 ctgcccaggg accaccgacc caccaccggg aggtaagctg gctgcctcgc gcgtttcggt 5641 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa 5701 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg 5761 ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atctagctag gtagctagag 5821 gatctttgtg aaggaacctt acttctgtgg tgtgacataa ttggacaaac tacctacaga 5881 gatttaaagc tctaaggtaa atataaaatt tttaagtgta taatgtgtta aactactgat 5941 tctaattgtt tgtgtatttt agattccaac ctatggaact gatgaatggg agcagtggtg 6001 gaatgccttt aatgaggaaa acctgttttg ctcagaagaa atgccatcta gtgatgatga 6061 ggctactgct gactctcaac attctactcc tccaaaaaag aagagaaagg tagaagaccc 6121 caaggacttt ccttcagaat tgctaagttt tttgagtcat gctgtgttta gtaatagaac 6181 tcttgcttgc tttgctattt acaccacaaa ggaaaaagct gcactgctat acaagaaaat 6241 tatggaaaaa tatttgatgt atagtgcctt gactagagat cataatcagc cataccacat 6301 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata 6361 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa 6421 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt 6481 tgtccaaact catcaatgta tcttatcatg tctggatcag cttcagaaga tggcggaggg 6541 cctccaacac agtaattttc ctcccgactc ttaaaataga aaatgtcaag tcagttaagc 6601 aggaagtgac taactgaccg cagctggccg tgcgacatcc tcttttaatt agttgctagg 6661 caactgccct ccagagggca gtgtggtttt gcaagaggaa gcaaaaagcc tctccaccca 6721 ggcctagaat gtttccaccc aatcattact atgacaacag ctgttttttt tagtattaag 6781 cagaggccgg gggcccctgg cctccgctta ctctggagaa aaagaagaga ggcattgtag 6841 aggcttccag aggcaacttg tcaaaacagg actgcttcta tttctgtcac actgtctggc 6901 cctgtcacaa ggtccagcac ctccataccc cctttaataa gcagtttggg aacgggtgcg 6961 ggtcttactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat 7021 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc 7081 cagaagtagt gaggaggctt ttttggaggc tgcattaatg aatcggccaa cgcgcgggga 7141 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 7201 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 7261 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 7321 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 7381 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 7441 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 7501 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 7561 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 7621 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 7681 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 7741 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta 7801 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 7861 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 7921 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 7981 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 8041 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 8101 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 8161 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 8221 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 8281 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 8341 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 8401 gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 8461 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 8521 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 8581 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 8641 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 8701 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 8761 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 8821 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 8881 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 8941 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 9001 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 9061 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca 9121 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg 9181 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag 9241 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg 9301 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg 9361 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc 9421 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cag //