LOCUS       Exported                7102 bp ds-DNA     circular SYN 04-ABR-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pD206-pLKO.1-TRC.mKO2_shHRASLS.1
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7102)
  AUTHORS   Douglas Adamoski
  TITLE     Direct Submission
  JOURNAL   Exported quarta-feira, 4 de abr de 2018 from SnapGene 2.8.3
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7102
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     misc_feature    516..521
                     /note="Kozak"
     CDS             519..1172
                     /codon_start=1
                     /product="monomeric Kusabira-Orange2 fluorescent protein"
                     /note="mKO2"
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     LTR             1274..1507
                     /note="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1579..1700
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      1740..1875
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     rep_origin      2082..2537
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2563..2667
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             2668..3528
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3699..4287
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     LTR             4983..5163
                     /note="5' LTR"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5210..5335
                     /note="HIV-1 psi"
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5828..6061
                     /note="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     promoter        6588..6828
                     /label=U6 Promoter
                     /note="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_feature    6839..6859
                     /note="HRASLS (57110) shRNA"
                     /note="shRNA TRCN0000378630
                     Target Sequence:CGATAAGTACCGTTGAGTTTG
                     Hairpin 
                     Sequence:5'-CCGG-CGATAAGTACCGTTGAGTTTG-CTCGAG-CAAACTCAACGGT
                     ACTTATCG-TTTTTG-3'"
     misc_feature    6866..6886
                     /note="HRASLS (57110) shRNA"
                     /note="shRNA TRCN0000378630
                     Target Sequence:CGATAAGTACCGTTGAGTTTG
                     Hairpin 
                     Sequence:5'-CCGG-CGATAAGTACCGTTGAGTTTG-CTCGAG-CAAACTCAACGGT
                     ACTTATCG-TTTTTG-3'"
     misc_feature    6933..7050
                     /note="cPPT/CTS"
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7099..507
                     /note="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
ORIGIN
        1 ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg
       61 cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt
      121 cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct
      181 ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa
      241 gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc
      301 cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc
      361 cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc
      421 gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt
      481 gaccgaatca ccgacctctc tccccagggg gatctaccat ggtgagcgtg atcaagcccg
      541 agatgaagat gaggtactac atggacggct ccgtcaatgg gcatgagttc acaatcgagg
      601 gtgagggcac aggcagacct tacgagggac atcaggagat gacactgcgc gtcacaatgg
      661 ccgagggcgg gccaatgcct ttcgccttcg acctggtgtc ccacgtgttc tgttacggcc
      721 acagagtttt taccaagtac ccagaagaga tcccagacta tttcaagcag gcctttcctg
      781 agggcctgtc ctgggagagg tccctggagt tcgaggacgg cggctccgcc tccgtgagcg
      841 cccacatcag cctgaggggc aacaccttct accacaagtc caagttcacc ggcgtgaact
      901 tccccgccga cggccccatc atgcagaacc agagcgtgga ctgggagccc tccaccgaga
      961 agatcaccgc cagcgacggc gtgctgaagg gcgacgtgac catgtacctg aagctggagg
     1021 gcggcggcaa ccacaagtgc cagatgaaga ccacctacaa ggccgccaag gagatcctgg
     1081 agatgcccgg cgaccactac atcggccaca ggctggtgag gaagaccgag ggcaacatca
     1141 ccgagcaggt ggaggacgcc gtggcccact ccggtggcgg aggctcgggc ggaggtgggt
     1201 cgggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     1261 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct
     1321 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     1381 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     1441 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     1501 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
     1561 atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     1621 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     1681 aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca
     1741 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     1801 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     1861 gcttttttgg aggcctaggg acgtacccaa ttcgccctat agtgagtcgt attacgcgcg
     1921 ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     1981 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     2041 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
     2101 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     2161 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     2221 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     2281 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     2341 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     2401 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     2461 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     2521 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     2581 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     2641 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     2701 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     2761 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     2821 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     2881 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     2941 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     3001 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     3061 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     3121 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     3181 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     3241 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     3301 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     3361 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     3421 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     3481 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     3541 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     3601 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     3661 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     3721 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     3781 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     3841 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     3901 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     3961 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     4021 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4081 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     4141 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     4201 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     4261 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     4321 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     4381 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     4441 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     4501 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     4561 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     4621 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     4681 agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg
     4741 gagctgcaag cttaatgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga
     4801 tgagttagca acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag
     4861 taaggtggta cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac
     4921 gaaccactga attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacataa
     4981 acgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     5041 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     5101 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     5161 gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg
     5221 caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac
     5281 gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat
     5341 taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa
     5401 aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa
     5461 tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc
     5521 ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg
     5581 tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga
     5641 gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag
     5701 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac
     5761 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag
     5821 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag
     5881 cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga
     5941 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca
     6001 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc
     6061 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta
     6121 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca
     6181 gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc
     6241 aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt
     6301 ttaacataac aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg
     6361 taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt
     6421 caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa
     6481 tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg aacggatctc
     6541 gacggtatcg atcacgagac tagcctcgag cggccgcccc cttcaccgag ggcctatttc
     6601 ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata attggaatta
     6661 atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc
     6721 ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa
     6781 cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccggcg
     6841 ataagtaccg ttgagtttgc tcgagcaaac tcaacggtac ttatcgtttt tgaattctcg
     6901 acctcgagac aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg
     6961 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta
     7021 caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca
     7081 ctttggccgc ggctcgaggg gg
//
