LOCUS       Exported                8255 bp DNA     circular SYN 10-MAR-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8255)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 16, 2022 from SnapGene 6.0.5
            https://www.snapgene.com
COMMENT     LOCUS       pLKO_TRC005             9335 bp    DNA     circular 
            29-JAN-2010
            DEFINITION  pLKO TRC v2 library vector, with 1.9 kb stuffer insert. 
            SOURCE
              ORGANISM
            COMMENT     This file is created by Vector NTI
                        http://www.invitrogen.com/
            COMMENT     ORIGDB|GenBank
            COMMENT     VNTDATE|488543517|
            COMMENT     VNTDBDATE|544461344|
            COMMENT     LSOWNER|
            COMMENT     VNTNAME|pLKO_TRC005|
            COMMENT     VNTAUTHORNAME| Sidi Chen
            COMMENT     VNTOAUTHORNAME|UNKNOWN|
            COMMENT     VNTREPLTYPE|Plasmid
FEATURES             Location/Qualifiers
     source          1..8255
                     /label=pLKO_TRC005
                     /organism="synthetic DNA construct"
     source          1..2353
                     /label=pLKO_TRC005
     LTR             1..413
                     /label=RSV/5'LTR
     misc_feature    464..601
                     /label=psi
     misc_feature    568..932
                     /label=gag
     misc_feature    1078..1319
                     /label=RRE
     misc_feature    1717..1965
                     /label=hU6 Promoter
                     /note="hU6 Promoter"
     primer_bind     1717..1736
                     /label=HKO035
                     /note="HKO035"
     misc_feature    1875..4750
                     /label=gB_pSC014
                     /note="gB_pSC014"
     misc_feature    1889..1910
                     /label=hU6P primer
                     /note="hU6P primer"
     misc_feature    1907..1966
                     /label=array_F
                     /note="array_F"
     misc_feature    1933..1965
                     /label=Readout F priming
                     /note="Readout F priming"
     misc_feature    1947..1966
                     /label=array_F_anneal
                     /note="array_F_anneal"
     misc_feature    1967..1972
                     /label=BsmbI
                     /note="BsmbI"
     misc_feature    1979..1984
                     /label=BsmbI
                     /note="BsmbI"
     misc_feature    1986..2121
                     /label=chimeric guide RNA scaffold
     misc_feature    1986..2121
                     /label=chimeric guide RNA scaffold
                     /note="chimeric guide RNA scaffold"
     misc_feature    1986..2009
                     /label=array_Rms2_anneal
                     /note="array_Rms2_anneal"
     misc_feature    1998..2031
                     /label=MS2 binding loop
     misc_feature    1998..2031
                     /label=MS2 binding loop
                     /note="MS2 binding loop"
     misc_feature    2046..2070
                     /label=Readout_Rms2_priming
                     /note="Readout_Rms2_priming"
     misc_feature    2068..2101
                     /label=MS2 binding loop
     misc_feature    2068..2101
                     /label=MS2 binding loop
                     /note="MS2 binding loop"
     misc_feature    2122..2128
                     /label=U6 terminator
     misc_feature    2122..2128
                     /label=U6 Terminator
                     /note="U6 Terminator"
     misc_feature    2184..2301
                     /label=cPPT
     misc_feature    2360..2381
                     /label=pCM111_R1
                     /note="pCM111_R1"
     promoter        2372..2627
                     /label=EFS-NS
                     /note="EFS-NS"
     misc_feature    2372..2398
                     /label=pCM111_R2, readout PCR2 priming site
                     /note="pCM111_R2, readout PCR2 priming site"
     misc_feature    2378..2397
                     /label=EFS-F1
                     /note="EFS-F1"
     misc_feature    2652..3137
                     /label=Thy1.1 mouse
                     /note="Thy1.1 mouse"
     misc_feature    2976..2978
                     /label=Valine syn mut GTC - GTG
                     /note="Valine syn mut GTC - GTG"
     misc_feature    3138..3590
                     /label=MS2
     misc_feature    3138..3200
                     /label=2A
     misc_feature    3138..3200
                     /label=P2A ?
                     /note="P2A ?"
     misc_feature    3201..3590
                     /label=MS2
                     /note="MS2"
     misc_feature    3366..3368
                     /label=N55K
     misc_feature    3591..3596
                     /label=AfeI
     misc_feature    3597..3641
                     /label=Linker (GGS)5
     misc_feature    3597..3641
                     /label=NLS
                     /note="NLS"
     misc_feature    3597..3641
                     /label=linker GGS 5 ?
                     /note="linker GGS 5 ?"
     misc_feature    3642..3674
                     /label=NLS
     misc_feature    3642..3674
                     /label=NLS ?
                     /note="NLS ?"
     misc_feature    3675..3680
                     /label=BamHI
     misc_feature    3681..4232
                     /label=p65
     misc_feature    3681..4232
                     /label=p65
                     /note="p65"
     misc_feature    4233..4247
                     /label=GGGGS
     misc_feature    4233..4247
                     /label=GGGGS
                     /note="GGGGS"
     misc_feature    4248..4619
                     /label=HSF1
     misc_feature    4248..4619
                     /label=HSF1
                     /note="HSF1"
     misc_feature    4620..4622
                     /label=stop
                     /note="stop"
     misc_feature    4635..4682
                     /label=Short polyA
                     /note="Short polyA"
     misc_feature    4691..4696
                     /label=mutate G - t to eliminate BsmBI site
                     /note="mutate G - t to eliminate BsmBI site"
     misc_feature    4693..4712
                     /label=pSC008_R1
                     /note="pSC008_R1"
     LTR             4771..5006
                     /label=3' LTR SIN
     polyA_signal    5083..5213
                     /label=SV40 polyA
     rep_origin      5560..5990
                     /direction=RIGHT
                     /label=f1 Ori
     misc_feature    6361..7167
                     /label=AmpR
     rep_origin      complement(7293..7908)
                     /direction=LEFT
                     /label=pUC Ori
ORIGIN
        1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
       61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
      121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
      181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
      241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
      421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
      481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
      541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
      601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
      661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
      721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
      781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
      841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
      901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
      961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     1681 tttcagaccc acctcccaac cccgagggga cccagagagg gcctatttcc catgattcct
     1741 tcatatttgc atatacgata caaggctgtt agagagataa ttagaattaa tttgactgta
     1801 aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt
     1861 gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat
     1921 ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccggaga cgggataccg
     1981 tctctgtttt agagctaggc caacatgagg atcacccatg tctgcagggc ctagcaagtt
     2041 aaaataaggc tagtccgtta tcaacttggc caacatgagg atcacccatg tctgcagggc
     2101 caagtggcac cgagtcggtg ctttttttgg atccaagctt ggcgtaacta gatcttgaga
     2161 caaatggcag tattcatcca caattttaaa agaaaagggg ggattggggg gtacagtgca
     2221 ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa
     2281 attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc actttggcgc
     2341 cggctcgagg gggcccgggg aattcgctag ctaggtcttg aaaggagtgg gaattggctc
     2401 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
     2461 ggtcggcaat tgatccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
     2521 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
     2581 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggacc ggttctagac
     2641 gtacggccac catgaaccca gccatcagcg tcgctctcct gctctcagtc ttgcaggtgt
     2701 cccgagggca gaaggtgacc agcctgacag cctgcctggt gaaccaaaac cttcgcctgg
     2761 actgccgcca tgagaataac accaaggata actccatcca gcatgagttc agcctgaccc
     2821 gagagaagag gaagcacgtg ctctcaggca cccttgggat acccgagcac acgtaccgct
     2881 cccgcgtcac cctctccaac cagccctata tcaaggtcct taccctagcc aacttcacca
     2941 ccaaggatga gggcgactac ttttgtgagc ttcgcgtgtc gggcgcgaat cccatgagct
     3001 ccaataaaag tatcagtgtg tatagagaca agctggtcaa gtgtggcggc ataagcctgc
     3061 tggttcagaa cacatcctgg atgctgctgc tgctgctttc cctctccctc ctccaagccc
     3121 tggacttcat ttctctgggc agtggagagg gcagaggaag tctgctaaca tgcggtgacg
     3181 tcgaggagaa tcctggccca atggcttcaa actttactca gttcgtgctc gtggacaatg
     3241 gtgggacagg ggatgtgaca gtggctcctt ctaatttcgc taatggggtg gcagagtgga
     3301 tcagctccaa ctcacggagc caggcctaca aggtgacatg cagcgtcagg cagtctagtg
     3361 cccagaagag aaagtatacc atcaaggtgg aggtccccaa agtggctacc cagacagtgg
     3421 gcggagtcga actgcctgtc gccgcttgga ggtcctacct gaacatggag ctcactatcc
     3481 caattttcgc taccaattct gactgtgaac tcatcgtgaa ggcaatgcag gggctcctca
     3541 aagacggtaa tcctatccct tccgccatcg ccgctaactc aggtatctac agcgctggag
     3601 gaggtggaag cggaggagga ggaagcggag gaggaggtag cggacctaag aaaaagagga
     3661 aggtggcggc cgctggatcc ccttcagggc agatcagcaa ccaggccctg gctctggccc
     3721 ctagctccgc tccagtgctg gcccagacta tggtgccctc tagtgctatg gtgcctctgg
     3781 cccagccacc tgctccagcc cctgtgctga ccccaggacc accccagtca ctgagcgctc
     3841 cagtgcccaa gtctacacag gccggcgagg ggactctgag tgaagctctg ctgcacctgc
     3901 agttcgacgc tgatgaggac ctgggagctc tgctggggaa cagcaccgat cccggagtgt
     3961 tcacagatct ggcctccgtg gacaactctg agtttcagca gctgctgaat cagggcgtgt
     4021 ccatgtctca tagtacagcc gaaccaatgc tgatggagta ccccgaagcc attacccggc
     4081 tggtgaccgg cagccagcgg ccccccgacc ccgctccaac tcccctggga accagcggcc
     4141 tgcctaatgg gctgtccgga gatgaagact tctcaagcat cgctgatatg gactttagtg
     4201 ccctgctgtc acagatttcc tctagtgggc agggaggagg tggaagcggc ttcagcgtgg
     4261 acaccagtgc cctgctggac ctgttcagcc cctcggtgac cgtgcccgac atgagcctgc
     4321 ctgaccttga cagcagcctg gccagtatcc aagagctcct gtctccccag gagcccccca
     4381 ggcctcccga ggcagagaac agcagcccgg attcagggaa gcagctggtg cactacacag
     4441 cgcagccgct gttcctgctg gaccccggct ccgtggacac cgggagcaac gacctgccgg
     4501 tgctgtttga gctgggagag ggctcctact tctccgaagg ggacggcttc gccgaggacc
     4561 ccaccatctc cctgctgaca ggctcggagc ctcccaaagc caaggacccc actgtctcct
     4621 aatgtacaag cgctaataaa agatctttat tttcattaga tctgtgtgtt ggttttttgt
     4681 gtggtaactc tagacgtgcg gtcgacttta agaccaatga cttacaaggc agctgtagat
     4741 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
     4801 caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag
     4861 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     4921 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     4981 tagtcagtgt ggaaaatctc tagcagtacg tatagtagtt catgtcatct tattattcag
     5041 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct
     5101 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5161 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag
     5221 ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     5281 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     5341 agctattcca gaagtagtga ggaggctttt ttggaggcct agggacgtac ccaattcgcc
     5401 ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga
     5461 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     5521 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     5581 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5641 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     5701 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     5761 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     5821 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5881 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     5941 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     6001 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
     6061 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     6121 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     6181 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     6241 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     6301 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     6361 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     6421 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     6481 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     6541 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     6601 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     6661 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     6721 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6781 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6841 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6901 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6961 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     7021 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     7081 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     7141 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     7201 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     7261 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     7321 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     7381 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     7441 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     7501 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     7561 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     7621 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     7681 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     7741 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     7801 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     7861 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     7921 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     7981 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     8041 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     8101 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     8161 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac
     8221 cctcactaaa gggaacaaaa gctggagctg caagc
//
