LOCUS       pRC0610_U6-SpCas9-XPR502-Puro-2A-tagBFP        9840 bp    DNA     circular     05-AUG-2022
DEFINITION  .
ACCESSION   urn.local...3h-e2233ii
KEYWORDS    .
SOURCE      
  ORGANISM  .
COMMENT     Created by Clontech Laboratories Inc. http://www.clontech.com Cat.
            No. 632565.
FEATURES             Location/Qualifiers
     LTR             1..635
                     /label="5'\LTR"
     source          1..9840
                     /dnas_title="4-0514-15_pSB700.U6.TTNsa.C3orf19Sp"
     misc_binding    636..653
                     /label="PBS"
     misc_feature    685..822
                     /label="packaging\signal"
     misc_binding    1303..1536
                     /label="RRE"
     misc_feature    2028..2151
                     /label="cPPT/CTS\(DNA\flap)"
     PCR_primer      2205..2225
                     /dnas_title="ATCGATCACGAGACTAGCCTC"
                     /pair="/primer=""ATCGATCACGAGACTAGCCTC"
                     /current=0
     promoter        2246..2486
                     /note="RNA polymerase III promoter for human U6 snRNA"
                     /label="U6 promoter"
     primer_bind     2246..2266
                     /note="Human U6 promoter, forward primer"
                     /label="hU6-F"
     source          <2246..>2726
                     /dnas_title="4-0514-15_pSB700.U6.TTNsa.C3orf19Sp"
     source          <2246..>2726
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          complement(<2246..>2726)
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     2417..2436
                     /note="Human U6 promoter, forward primer"
                     /label="LKO.1 5'"
     PCR_primer      2446..2495
                     /created_by="charir2"
                     /label="Forward-flank"
     PCR_primer      2473..2495
                     /created_by="charir2"
                     /label="Primer-592"
     PCR_primer      2473..2493
                     /created_by="charir2"
                     /label="F"
     unsure          2496..2501
                     /created_by="charir2"
                     /label="BsmbI"
     misc_feature    2502..2654
                     /Original_Bases="TGACCGTCTCTGTTTAAGAGCTATGCTGGAAA"
                     /label="TGACCGTCTCTGTTTAAGAGCTATGCTGGAAA"
                     /note="Geneious type: Editing History Replacement"
     unsure          2514..2519
                     /created_by="charir2"
                     /label="Bsmbi"
     misc_RNA        2521..2726
                     /created_by="charir2"
                     /label="2X-PP7-2X-MS2"
     PCR_primer      complement(2759..2777)
                     /dnas_title="GGGCTATGAACTAATGACG"
                     /pair="/primer=""GGGCTATGAACTAATGACG"
                     /current=0
     enhancer        2797..3084
                     /note="%22CAG enhancer%22 H CMV IE enhancer like pCAGGS"
     PCR_primer      complement(2843..2865)
                     /created_by="charir2"
                     /label=2402
     promoter        3103..4343
                     /note="CAG promoter like pCAGGS"
     misc_feature    4419..4424
                     /note="Kozak"
     CDS             4425..5021
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
                     /label="PuroR"
     primer_bind     complement(5001..5051)
                     /ApEinfo_revcolor="#a020f0"
                     /ApEinfo_fwdcolor="#a020f0"
                     /label="FSR-51 (Puro Rev + 30bp of P2A Homology)"
     primer_bind     5022..5072
                     /ApEinfo_revcolor="#ff0000"
                     /ApEinfo_fwdcolor="#ff0000"
                     /label="FSR-52 (P2A Fwd + 30bp FSR-51 homology)"
     CDS             5037..5093
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
                     /label="P2A"
     misc_feature    5084
                     /Original_Bases="C"
                     /label="C"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    5087
                     /Original_Bases="C"
                     /label="C"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    5090
                     /Original_Bases="C"
                     /label="C"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    5093
                     /Original_Bases="C"
                     /label="C"
                     /note="Geneious type: Editing History Replacement"
     CDS             5094..>5792
                     /kind="reporter"
                     /note="Blue fluorescent reporter protein - Ex:402, Em:457"
                     /Color="#3366ff"
                     /product="TagBFP"
                     /gene="TagBFP"
                     /label="TagBFP"
     misc_feature    5384
                     /Original_Bases="A"
                     /label="A"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    5528
                     /Original_Bases="G"
                     /label="G"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    5531
                     /Original_Bases="G"
                     /label="G"
                     /note="Geneious type: Editing History Replacement"
     CDS             <5793..5795
                     /ApEinfo_revcolor="#ff0000"
                     /ApEinfo_fwdcolor="#ff0000"
                     /label="TagRFP"
     misc_feature    5808..6399
                     /label="WPRE"
     LTR             6602..7238
                     /label="3'\LTR"
     misc_feature    7414..7443
                     /note="lac promoter"
     rep_origin      complement(7708..8381)
                     /label="pUC\ori"
     CDS             complement(8526..9522)
                     /label="Amp(R) with promoter 7449-8379"
     misc_feature    9542..9673
                     /note="SV40 polyA"
ORIGIN      
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cattagtgaa cggatctcga cggtatcgat cacgagacta
     2221 gcctcgagcg gccgccccct tcaccgaggg cctatttccc atgattcctt catatttgca
     2281 tatacgatac aaggctgtta gagagataat tagaattaat ttgactgtaa acacaaagat
     2341 attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa
     2401 attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt
     2461 ggctttatat atcttgtgga aaggacgaaa caccggagac gaccggtttt ttgcgtctct
     2521 gtttaagagc tatgctgcga atacgaggga gcagacgata tggcgtcgct cctctccacg
     2581 agagcatatg ggctccgtgg tctcgtacac catcagggta cgtatcagac accatcaggg
     2641 tctgctcgta ttcgcagcat agcaagttta aataaggcta gtccgttatc aacttgaaaa
     2701 agtggcaccg agtcggtgct ttttttgaat tctcgacctc gagacaaatg gcagtattcg
     2761 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     2821 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     2881 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     2941 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     3001 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     3061 cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc cacgttctgc
     3121 ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa
     3181 ttattttgtg cagcgatggg ggcggggggg ggggggcgcg cgccaggcgg ggcggggcgg
     3241 ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg
     3301 ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc
     3361 gcgcggcggg cggggagtcg ctgcgacgct gccttcgccc cgtgccccgc tccgccgccg
     3421 cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg
     3481 acggcccttc tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc
     3541 tgtggctgcg tgaaagcctt gaggggctcc gggagggccc tttgtgcggg gggagcggct
     3601 cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg
     3661 gcggctgtga gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg
     3721 ggagcgcggc cgggggcggt gccccgcggt gcgggggggg ctgcgagggg aacaaaggct
     3781 gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc
     3841 aaccccccct gcacccccct ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc
     3901 tccgtacggg gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg caggtggggg
     3961 tgccgggcgg ggcggggccg cctcgggccg gggagggctc gggggagggg cgcggcggcc
     4021 cccggagcgc cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat
     4081 cgtgcgagag ggcgcaggga cttcctttgt cccaaatctg tgcggagccg aaatctggga
     4141 ggcgccgccg caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg
     4201 aaatgggcgg ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc
     4261 ctcggggctg tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg
     4321 gcttctggcg tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct
     4381 ttttcctaca gctcctgggc aacgtgctgg ttattgtggc caccatgacc gagtacaagc
     4441 ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg
     4501 cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg
     4561 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg
     4621 tcgcggacga cggcgccgcc gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
     4681 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
     4741 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
     4801 ccaccgtcgg agtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc
     4861 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
     4921 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
     4981 aaggaccgcg cacctggtgc atgacccgca agcccggtgc cgctagcggc agcggcgcca
     5041 ccaacttcag cctgctgaag caggccggcg acgtggagga gaatcctgga ccgatgagcg
     5101 agctgattaa ggagaacatg cacatgaagc tgtacatgga gggcaccgtg gacaaccatc
     5161 acttcaagtg cacatccgag ggcgaaggca agccctacga gggcacccag accatgagaa
     5221 tcaaggtggt cgagggcggc cctctcccct tcgccttcga catcctggct actagcttcc
     5281 tctacggcag caagaccttc atcaaccaca cccagggcat ccccgacttc ttcaagcagt
     5341 ccttccctga gggcttcaca tgggagagag tcaccacata cgaggacggg ggcgtgctga
     5401 ccgctaccca ggacaccagc ctccaggacg gctgcctcat ctacaacgtc aagatcagag
     5461 gggtgaactt cacatccaac ggccctgtga tgcagaagaa aacactcggc tgggaggcct
     5521 tcaccgaaac actgtacccc gctgacggcg gcctggaagg cagaaacgac atggccctga
     5581 agctcgtggg cgggagccat ctgatcgcaa acatcaagac cacatataga tccaagaaac
     5641 ccgctaagaa cctcaagatg cctggcgtct actatgtgga ctacagactg gaaagaatca
     5701 aggaggccaa caacgagacc tacgtcgagc agcacgaggt ggcagtggcc agatactgcg
     5761 acctccctag caaactgggg cacaagctta attgaagcgt ctggaacaat caacctctgg
     5821 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     5881 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     5941 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     6001 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     6061 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     6121 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     6181 attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca
     6241 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     6301 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     6361 agacgagtcg gatctccctt tgggccgcct ccccgcctgg aattaattct gcagtcgaga
     6421 cctagaaaaa catggagcaa tcacaagtag caatacagca gctaccaatg ctgattgtgc
     6481 ctggctagaa gcacaagagg aggaggaggt gggttttcca gtcacacctc aggtaccttt
     6541 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagagggg
     6601 actggaaggg ctaattcact cccaacgaag acaagatatc cttgatctgt ggatctacca
     6661 cacacaaggc tacttccctg attagcagaa ctacacacca gggccagggg tcagatatcc
     6721 actgaccttt ggatggtgct acaagctagt accagttgag ccagataagg tagaagaggc
     6781 caataaagga gagaacacca gcttgttaca ccctgtgagc ctgcatggga tggatgaccc
     6841 ggagagagaa gtgttagagt ggaggtttga cagccgccta gcatttcatc acgtggcccg
     6901 agagctgcat ccggagtact tcaagaactg ctgatatcga gcttgctaca agggactttc
     6961 cgctggggac tttccaggga ggcgtggcct gggcgggact ggggagtggc gagccctcag
     7021 atcctgcata taagcagctg ctttttgcct gtactgggtc tctctggtta gaccagatct
     7081 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     7141 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     7201 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     7261 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggcct tgacattgct
     7321 agcgttttac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     7381 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     7441 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     7501 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     7561 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     7621 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     7681 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     7741 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     7801 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     7861 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7921 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     7981 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     8041 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     8101 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     8161 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     8221 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     8281 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     8341 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     8401 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     8461 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     8521 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     8581 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     8641 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     8701 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     8761 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     8821 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     8881 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     8941 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     9001 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     9061 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     9121 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     9181 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     9241 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     9301 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     9361 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     9421 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     9481 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatcaa
     9541 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     9601 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     9661 tcatgtctgg atcaactgga taactcaagc taaccaaaat catcccaaac ttcccacccc
     9721 ataccctatt accactgcca attacctgtg gtttcattta ctctaaacct gtgattcctc
     9781 tgaattattt tcattttaaa gaaattgtat ttgttaaata tgtactacaa acttagtagt
//
