LOCUS Exported 6699 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pMKO.1 puro Rb shRNA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6699) AUTHORS Boehm JS, Hession MT, Bulmer SE, Hahn WC TITLE Transformation of human and murine fibroblasts without viral oncoproteins. JOURNAL Mol Cell Biol. 2005 Aug . 25(15):6464-74. PUBMED 16024784 REFERENCE 2 (bases 1 to 6699) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6699 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 116..134 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" enhancer 234..613 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 614..817 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 768..788 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 818..993 /label=5' LTR (truncated) /note="truncated long terminal repeat from Moloney murine sarcoma virus" misc_feature 1056..1255 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 1456..1872 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1802..1824 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1840..1856 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" promoter 1908..2148 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 1908..1928 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 2079..2098 /label=LKO.1 5' /note="Human U6 promoter, forward primer" promoter 2246..2575 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2426..2561 /label=SV40 ori /note="SV40 origin of replication" CDS 2585..3184 /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(2585..2604) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 3081..3101 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 3380..3805 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from Moloney murine leukemia virus" primer_bind complement(3908..3930) /label=pGEX 3' /note="pGEX vectors, reverse primer" promoter 4071..4400 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4251..4386 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4422..4441 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" rep_origin complement(4813..5401) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4893..4912) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5572..6432) /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 6195..6214 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6433..6537) /gene="bla" /label=AmpR promoter ORIGIN 1 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 61 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 121 gcgtatcacg aggccctttc gtcttcaaga attagcttgg ccattgcata cgttgtatcc 181 atatcataat atgtacattt atattggctc atgtccaaca ttaccgccat gttgacattg 241 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat 301 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc 361 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca 421 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta 481 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta 541 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat 601 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga 661 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca 721 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg 781 taggcgtgta cggtgggagg tctatataag cagagctcaa taaaagagcc cacaacccct 841 cactcggcgc gccagtcttc cgatagactg cgtcgcccgg gtacccgtat tcccaataaa 901 gcctcttgct gtttgcatcc gaatcgtggt ctcgctgttc cttgggaggg tctcctctga 961 gtgattgact acccacgacg ggggtctttc atttgggggc tcgtccggga tttggagacc 1021 cctgcccagg gaccaccgac ccaccaccgg gaggtaagct ggccagcaac ttatctgtgt 1081 ctgtccgatt gtctagtgtc tatgtttgat gttatgcgcc tgcgtctgta ctagttagct 1141 aactagctct gtatctggcg gacccgtggt ggaactgacg agttctgaac acccggccgc 1201 aaccctggga gacgtcccag ggactttggg ggccgttttt gtggcccgac ctgaggaagg 1261 gagtcgatgt ggaatccgac cccgtcagga tatgtggttc tggtaggaga cgagaaccta 1321 aaacagttcc cgcctccgtc tgaatttttg ctttcggttt ggaaccgaag ccgcgcgtct 1381 tgtctgctgc agcgctgcag catcgttctg tgttgtctct gtctgactgt gtttctgtat 1441 ttgtctgaaa attagggcca gactgttacc actcccttaa gtttgacctt aggtcactgg 1501 aaagatgtcg agcggatcgc tcacaaccag tcggtagatg tcaagaagag acgttgggtt 1561 accttctgct ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga cggcaccttt 1621 aaccgagacc tcatcaccca ggttaagatc aaggtctttt cacctggccc gcatggacac 1681 ccagaccagg tcccctacat cgtgacctgg gaagccttgg cttttgaccc ccctccctgg 1741 gtcaagccct ttgtacaccc taagcctccg cctcctcttc ctccatccgc cccgtctctc 1801 ccccttgaac ctcctcgttc gaccccgcct cgatcctccc tttatccagc cctcactcct 1861 tctctaggcg ccggaattga agatctgtcg acaaggtcgg gcaggaagag ggcctatttc 1921 ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata attagaatta 1981 atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc 2041 ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa 2101 cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccggca 2161 gagatcgtgt attgagattc tcgagaatct caatacacga tctctgtttt tgaattcgcc 2221 agcacagtgg tcgaccctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctt 2281 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa 2341 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac 2401 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc 2461 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc 2521 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct 2581 taccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc 2641 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc 2701 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct 2761 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc 2821 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag 2881 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa 2941 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct 3001 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt 3061 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac 3121 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc 3181 ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc 3241 gattagtcca atttgttaaa gacaggatat cagtggtcca ggctctagtt ttgactcaac 3301 aatatcacca gctgaagcct atagagtacg agccatagat aaaataaaag attttattta 3361 gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag 3421 taacgccatt ttgcaaggca tggaaaaata cataactgag aatagagaag ttcagatcaa 3481 ggtcaggaac agatggaaca gggtcgaccc tagagaacca tcagatgttt ccagggtgcc 3541 ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct 3601 tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggg 3661 gcgccagtcc tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg 3721 cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac 3781 tacccgtcag cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca 3841 gggaccaccg acccaccacc gggaggtaag ctggctgcct cgcgcgtttc ggtgatgacg 3901 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg 3961 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag 4021 ccatgaccca gtcacgtagc gatagcggag tgtagatccg gctgtggaat gtgtgtcagt 4081 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 4141 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 4201 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc 4261 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg 4321 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg 4381 gaggcctagg cttttgcaaa aagcttactg gcttaactat gcggcatcag agcagattgt 4441 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg 4501 catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 4561 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 4621 cgcaggaaag aacatggctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 4681 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 4741 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 4801 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 4861 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 4921 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 4981 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 5041 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 5101 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 5161 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 5221 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct 5281 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 5341 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 5401 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 5461 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 5521 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 5581 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 5641 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 5701 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 5761 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 5821 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 5881 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 5941 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 6001 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 6061 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 6121 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 6181 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 6241 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 6301 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 6361 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 6421 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 6481 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 6541 atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta 6601 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttaa 6661 ttattgaagc atttatcagg gttattgtct catgagcgg //