LOCUS Exported 6368 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pMKO.1 puro p53 shRNA 1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6368) AUTHORS Masutomi K, Yu EY, Khurts S, Ben-Porath I, Currier JL, Metz GB, Brooks MW, Kaneko S, Murakami S, DeCaprio JA, Weinberg RA, Stewart SA, Hahn WC TITLE Telomerase maintains telomere structure in normal human cells. JOURNAL Cell. 2003 Jul 25. 114(2):241-53. PUBMED 12887925 REFERENCE 2 (bases 1 to 6368) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6368 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..241 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 1..21 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 172..191 /label=LKO.1 5' /note="Human U6 promoter, forward primer" promoter 338..667 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 518..653 /label=SV40 ori /note="SV40 origin of replication" CDS 677..1276 /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(677..696) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 1173..1193 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 1472..1897 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from Moloney murine leukemia virus" primer_bind complement(2000..2022) /label=pGEX 3' /note="pGEX vectors, reverse primer" promoter 2163..2492 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2343..2478 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2514..2533 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(2614..2631) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2785..3373) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2865..2884) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3544..4404) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 4167..4186 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4405..4509) /gene="bla" /label=AmpR promoter primer_bind 4577..4595 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" enhancer 4695..5074 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 5075..5278 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5229..5249 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 5279..5454 /label=5' LTR (truncated) /note="truncated long terminal repeat from Moloney murine sarcoma virus" misc_feature 5517..5716 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 5917..6333 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 6263..6285 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 6301..6317 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" ORIGIN 1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag 61 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga 121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat 181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga 241 cgaaacaccg gtgactccag tggtaatcta cttcaagaga gtagattaca ctggagtctt 301 tttgaattcg ccagcacagt ggtcgaccct gtggaatgtg tgtcagttag ggtgtggaaa 361 gtccccaggc ttcccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac 421 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa 481 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag 541 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc 601 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt 661 ttgcaaaaag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga 721 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca 781 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac 841 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt 901 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat 961 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc 1021 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca 1081 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg 1141 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg 1201 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg 1261 caagcccggt gcctgacgcc cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg 1321 accccatgca tcgattagtc caatttgtta aagacaggat atcagtggtc caggctctag 1381 ttttgactca acaatatcac cagctgaagc ctatagagta cgagccatag ataaaataaa 1441 agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca 1501 agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaga 1561 agttcagatc aaggtcagga acagatggaa cagggtcgac cctagagaac catcagatgt 1621 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt 1681 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac 1741 ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa 1801 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc 1861 tgagtgattg actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga 1921 gacccctgcc cagggaccac cgacccacca ccgggaggta agctggctgc ctcgcgcgtt 1981 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc 2041 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt 2101 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtagatc cggctgtgga 2161 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 2221 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca 2281 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc 2341 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt 2401 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag 2461 gaggcttttt tggaggccta ggcttttgca aaaagcttac tggcttaact atgcggcatc 2521 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 2581 gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 2641 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 2701 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 2761 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 2821 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 2881 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 2941 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 3001 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 3061 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 3121 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 3181 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat 3241 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 3301 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa 3361 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga 3421 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct 3481 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga 3541 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc 3601 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg 3661 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat 3721 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat 3781 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg 3841 caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc 3901 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa 3961 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc 4021 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt 4081 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag 4141 ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt 4201 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag 4261 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac 4321 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc 4381 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca 4441 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg 4501 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat 4561 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcaag aattagcttg 4621 gccattgcat acgttgtatc catatcataa tatgtacatt tatattggct catgtccaac 4681 attaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc 4741 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc 4801 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt 4861 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca 4921 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg 4981 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca 5041 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa 5101 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa 5161 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc 5221 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctca 5281 ataaaagagc ccacaacccc tcactcggcg cgccagtctt ccgatagact gcgtcgcccg 5341 ggtacccgta ttcccaataa agcctcttgc tgtttgcatc cgaatcgtgg tctcgctgtt 5401 ccttgggagg gtctcctctg agtgattgac tacccacgac gggggtcttt catttggggg 5461 ctcgtccggg atttggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc 5521 tggccagcaa cttatctgtg tctgtccgat tgtctagtgt ctatgtttga tgttatgcgc 5581 ctgcgtctgt actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac 5641 gagttctgaa cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt 5701 tgtggcccga cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt 5761 ctggtaggag acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt 5821 tggaaccgaa gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc 5881 tgtctgactg tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta 5941 agtttgacct taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat 6001 gtcaagaaga gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga 6061 tggccgcgag acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt 6121 tcacctggcc cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg 6181 gcttttgacc cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt 6241 cctccatccg ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc 6301 ctttatccag ccctcactcc ttctctaggc gccggaattg aagatctgtc gacaaggtcg 6361 ggcaggaa //