LOCUS Exported 6540 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses 3xFNLDD in mammalian cells. ACCESSION . VERSION . KEYWORDS 3xFNLDD/pMXs-puro SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6540) AUTHORS Fujita T, Asano Y, Ohtsuka J, Takada Y, Saito K, Ohki R, Fujii H TITLE Identification of telomere-associated molecules by engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP). JOURNAL Sci Rep. 2013 Nov 8;3:3171. doi: 10.1038/srep03171. PUBMED 24201379 REFERENCE 2 (bases 1 to 6540) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6540 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 22..615 /note="long terminal repeat from Moloney murine leukemia virus" misc_feature 678..1035 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 1094..1510 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1440..1462 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1478..1494 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" misc_feature 1520..1895 /label=pol region /note="Moloney murine leukemia virus (MMLV) pol region containing the splice acceptor site" primer_bind 1832..1848 /label=pBMN 5' /note="MMLV sequence, for inserts in pBMN retroviral vector" CDS 1973..2038 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 2075..2095 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 2108..2716 /codon_start=1 /gene="lexA" /product="E. coli DNA-binding protein that represses genes involved in the SOS response to DNA damage" /label=LexA /note="The N-terminal DNA-binding domain recognizes the 16-bp palindromic sequence 5'-CTGTATATATATACAG-3'." /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAAGEPLLAQQHIEGHYQVDPSL FKPNADFLLRVSGMSMKDIGIMDGDLLAVHKTQDVRNGQVVVARIDDEVTVKRLKKQGN KVELLPENSEFKPIVVDLRQQSFTIEGLAVGVIRNGDWL" primer_bind 2651..2672 /label=LexA /note="LexA DNA binding domain, forward primer" primer_bind complement(2875..2895) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2880..3209 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3060..3195 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3122..3141 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3219..3818 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(3219..3238) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 3715..3735 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 4045..4514 /note="long terminal repeat from Moloney murine leukemia virus" primer_bind complement(4644..4661) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4815..5403) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4895..4914) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5567..6427) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 6190..6209 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6428..6532) /gene="bla" /label=AmpR promoter ORIGIN 1 cccgaaaagt gccacctgca taatgaaaga ccccacctgt aggtttggca agctagctta 61 agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaaa agttcagatc 121 aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag 181 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat 241 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg 301 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct 361 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg 421 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc 481 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc 541 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag 601 cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg 661 acccaccacc gggaggtaag ctggccagca acttatctgt gtctgtccga ttgtctagtg 721 tctatgactg attttatgcg cctgcgtcgg tactagttag ctaactagct ctgtatctgg 781 cggacccgtg gtggaactga cgagttcgga acacccggcc gcaaccctgg gagacgtccc 841 agggacttcg ggggccgttt ttgtggcccg acctgagtcc aaaaatcccg atcgttttgg 901 actctttggt gcacccccct aataggaggg atatgtggtt ctggtaggag acgagaacct 961 aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tgggaccgaa gccgcgccgc 1021 gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt 1081 gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa 1141 agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac 1201 cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa 1261 ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc 1321 agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt 1381 caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc 1441 ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc 1501 tctaggcgcc cccatatggc catatgagat cttatatggg gcacccccgc cccttgtaaa 1561 cttccctgac cctgacatga caagagttac taacagcccc tctctccaag ctcacttaca 1621 ggctctctac ttagtccagc acgaagtctg gagacctctg gcggcagcct accaagaaca 1681 actggaccga ccggtggtac ctcaccctta ccgagtcggc gacacagtgt gggtccgccg 1741 acaccagact aagaacctag aacctcgctg gaaaggacct tacacagtcc tgctgaccac 1801 ccccaccgcc ctcaaagtag acggcatcgc agcttggata cacgccgccc acgtgaaggc 1861 tgccgacccc gggggtggac ccatcctcta gactgccgga tctagctagt taattaagga 1921 tcccagtgtg gtggtaggga attcgtttag tgaaccgtca gaattaacca tggactacaa 1981 agaccatgac ggtgattata aagatcatga catcgattac aaggatgacg atgacaagct 2041 tgcggccgcg aattcatcga tagatcgatc ccctccaaaa aagaagagaa aggtcgaacc 2101 tgagggaatg aaagcgttaa cggccaggca acaagaggtg tttgatctca tccgtgatca 2161 catcagccag acaggtatgc cgccgacgcg tgcggaaatc gcgcagcgtt tggggttccg 2221 ttccccaaac gcggctgaag aacatctgaa ggcgctggca cgcaaaggcg ttattgaaat 2281 tgtttccggc gcatcacgcg ggattcgtct gttgcaggaa gaggaagaag ggttgccgct 2341 ggtaggtcgt gtggctgccg gtgaaccact tctggcgcaa cagcatattg aaggtcatta 2401 tcaggtcgat ccttccttat tcaagccgaa tgctgatttc ctgctgcgcg tcagcgggat 2461 gtcgatgaaa gatatcggca ttatggatgg tgacttgctg gcagtgcata aaactcagga 2521 tgtacgtaac ggtcaggtcg ttgtcgcacg tattgatgac gaggttaccg ttaagcgcct 2581 gaaaaaacag ggcaataaag tcgaactgtt gccagaaaat agcgagttta aaccaattgt 2641 cgtagatctt cgtcagcaga gcttcaccat tgaagggctg gcggttgggg ttattcgcaa 2701 cggcgactgg ctgtagtcga aattccagct gagcgccggt cgctaccatt accagttggt 2761 ctggtgtcaa aaataataat aaccgggcag gccatgtctg cccgtatttc gcgtaaggaa 2821 atccattatg tactatttaa actcgagcgg ccgccagcac agtggtcgac ctgtggaatg 2881 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 2941 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa 3001 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca 3061 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt 3121 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag 3181 gcttttttgg aggcctaggc ttttgcaaaa agcttaccat gaccgagtac aagcccacgg 3241 tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg 3301 ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg 3361 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg 3421 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt 3481 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac 3541 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg 3601 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag 3661 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc 3721 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac 3781 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg cccgccccac gacccgcagc 3841 gcccgaccga aaggagcgca cgaccccatg catcgtcgac gataaaataa aagattttat 3901 ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt 3961 aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagag aagttcagat 4021 caaggtcagg aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca 4081 gttcctgccc cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga 4141 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc 4201 ggtccagccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc 4261 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc 4321 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc 4381 ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat 4441 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca 4501 gcgggggtct ttcacatgca gcatgtatca aaattaattt ggtttttttt cttaagtatt 4561 tacattaaat ggccatagtt gcattaatga atcggccaac gcgcggggag aggcggtttg 4621 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg 4681 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat 4741 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc 4801 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc 4861 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga 4921 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt 4981 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg 5041 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc 5101 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg 5161 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc 5221 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg 5281 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc 5341 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct 5401 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt 5461 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttgcggccg 5521 gccgcaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa 5581 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc 5641 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga 5701 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa 5761 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 5821 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 5881 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 5941 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg 6001 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 6061 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 6121 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 6181 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 6241 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 6301 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 6361 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 6421 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 6481 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc //