LOCUS       pUSEPR                  7877 bp ds-DNA     circular     18-NOV-2021
DEFINITION  .
ACCESSION   urn.local...2od-btutnh0
VERSION     urn.local...2od-btutnh0
KEYWORDS    "vector backbone:pLKO" "insert" "Sequence" "source:Francisco;
            Sanches-Rivera - Scott Lowe lab" "date;
            generated/aquired:2018-09-17" "antibiotic resistance:Amp";
            "bacterial strain for transformation:Stbl2, Stbl3" "bacterial;
            glycerol stock available?:yes, in stbl2" "creator:Francisco J.
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     regulatory      237..616
                     /note="Geneious type: enhancer"
                     /standard_name="CMV enhancer"
     regulatory      618..816
                     /note="Geneious type: promoter"
                     /standard_name="CMV promoter"
     misc_feature    834..1014
                     /note="Geneious type: LTR"
                     /standard_name="5' LTR (truncated)"
     misc_feature    1061..1186
                     /standard_name="HIV-1 Ψ"
     misc_feature    1685..1918
                     /standard_name="RRE"
     misc_feature    2138..2157
                     /standard_name="lenti_seq_f"
     misc_feature    2445..2562
                     /standard_name="cPPT/CTS"
     misc_feature    2625..2654
                     /standard_name="pAF Gibson"
     regulatory      2655..2895
                     /note="Geneious type: promoter"
                     /standard_name="U6 promoter"
     primer_bind     2655..2675
                     /standard_name="U6-Scaffold Fwd Primer (w/o gibson
                     Adapters)"
     misc_feature    2881..2900
                     /note="Geneious type: PCR_primer"
                     /standard_name="F"
     unsure          2882..3142
     misc_feature    2882..2904
                     /note="Geneious type: PCR_primer"
                     /standard_name="Mix"
     misc_feature    2900..3013
                     /standard_name="Improved sgRNA Scaffold (BsmBI)"
     misc_feature    2900..2925
                     /standard_name="Benedikt BsmBI Filler"
     misc_feature    2922..2971
                     /note="Geneious type: PCR_primer"
                     /standard_name="end"
     misc_feature    2922..2939
                     /note="Geneious type: PCR_primer"
                     /standard_name="F"
     misc_feature    2926..2946
                     /standard_name="Modified Stem Loop"
     misc_feature    complement(2949..2971)
                     /note="Geneious type: PCR_primer"
                     /standard_name="R"
     primer_bind     complement(2993..3013)
                     /standard_name="U6-Scaffold Rev Primer (w/o gibson
                     Adapters)"
     misc_feature    3014..3043
                     /standard_name="pBF Gibson"
     misc_feature    complement(3014..3043)
                     /standard_name="gAR"
     primer_bind     3044..3064
                     /standard_name="FSR-19 (w/o Gibson Adapters)"
     misc_feature    complement(3045..3064)
                     /note="Geneious type: PCR_primer"
                     /standard_name="R"
     regulatory      3069..3280
                     /note="Geneious type: promoter"
                     /standard_name="EF-1α core promoter"
     misc_feature    complement(3123..3142)
                     /standard_name="Lenti_Seq_r"
     misc_feature    complement(3170..3189)
                     /note="Geneious type: PCR_primer"
                     /standard_name="R"
     primer_bind     complement(3274..3294)
                     /standard_name="FSR-20 (w/o Gibson Adapters)"
     primer_bind     3295..3351
                     /standard_name="FSR-50 (Full Primer)"
     primer_bind     3295..3351
                     /standard_name="Puro Fwd + gBF"
     misc_feature    3295..3324
                     /standard_name="gBF Adapter"
     misc_feature    3325..3330
                     /standard_name="KOZAK"
     CDS             3331..3927
                     /standard_name="PuroR"
     primer_bind     complement(3907..3957)
                     /standard_name="FSR-51 (Puro Rev + 30bp of P2A Homology)"
     primer_bind     3928..3978
                     /standard_name="FSR-52 (P2A Fwd + 30bp FSR-51 homology)"
     CDS             3943..3999
                     /standard_name="P2A"
     unsure          complement(3980..3999)
     CDS             4000..4713
                     /standard_name="TagRFP"
     primer_bind     4000..4020
                     /standard_name="FSR-48 (w/o Gibson Adapters)"
     exon            4300..4305
                     /standard_name="Recode"
     misc_feature    complement(4689..4710)
                     /note="Geneious type: PCR_primer"
                     /standard_name="R"
     primer_bind     complement(4693..4743)
                     /standard_name="FSR-49 (Full Primer)"
     misc_feature    complement(4714..4743)
                     /standard_name="gBR (Rev Comp)"
     misc_feature    4766..5354
                     /standard_name="WPRE"
     primer_bind     complement(5357..5373)
                     /standard_name="KS primer"
     misc_feature    5883..6063
                     /note="Geneious type: LTR"
                     /standard_name="5' LTR (truncated)"
     rep_origin      complement(6125..6713)
     CDS             complement(6884..7675)
                     /standard_name="AmpR"
     CDS             complement(7676..7744)
                     /standard_name="AmpR"
     regulatory      complement(7745..7849)
                     /note="Geneious type: promoter"
                     /standard_name="AmpR promoter"
ORIGIN      
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc
      841 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      901 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      961 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
     1021 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
     1081 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
     1141 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
     1201 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
     1261 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
     1321 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
     1381 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
     1441 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
     1501 aagtaagacc accgcacagc aagcggccgg ccgcgctgat cttcagacct ggaggaggag
     1561 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
     1621 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
     1681 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
     1741 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
     1801 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
     1861 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
     1921 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
     1981 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
     2041 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
     2101 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
     2161 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
     2221 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
     2281 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
     2341 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac
     2401 tgcgtgcgcc aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg
     2461 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
     2521 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac
     2581 agcagagatc cagtttggtt agtaccgggc ccgggtcgac tagtgatcag tgtgagggag
     2641 tgtaaagctg gtttgagggc ctatttccca tgattccttc atatttgcat atacgataca
     2701 aggctgttag agagataatt agaattaatt tgactgtaaa cacaaagata ttagtacaaa
     2761 atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta
     2821 aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata
     2881 tcttgtggaa aggacgaaac accggagacg tgaccgtctc tgtttaagag ctatgctgga
     2941 aacagcatag caagtttaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag
     3001 tcggtgcttt tttaaaccgc tgttcctagg aatcccgagg ccttcgagtg gctccggtgc
     3061 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
     3121 caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
     3181 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
     3241 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtgtcgtgac gcgggacccg
     3301 acattagcgc tacagcttaa gcgggccacc atgaccgagt acaagcccac ggtgcgcctc
     3361 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     3421 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     3481 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     3541 gccgccgtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     3601 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     3661 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggagtc
     3721 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     3781 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     3841 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     3901 tggtgcatga cccgcaagcc cggtgccgct agcggcagcg gcgccaccaa cttcagcctg
     3961 ctgaagcagg ccggcgacgt ggaggagaac cccggcccca tggtgtctaa gggcgaagag
     4021 ctgattaagg agaacatgca catgaagctg tatatggagg gcaccgtgaa caaccaccac
     4081 ttcaagtgca catccgaggg cgaaggcaag ccctacgagg gcacccagac catgagaatc
     4141 aaggtggtcg agggcggccc tctccccttc gccttcgaca tcctggctac cagcttcatg
     4201 tacggcagca gaaccttcat caaccacacc cagggcatcc ccgacttctt taagcagtcc
     4261 ttccctgagg gcttcacatg ggagagagtc accacatacg aagacggggg cgtgctgacc
     4321 gctacccagg acaccagcct ccaggacggc tgcctcatct acaacgtcaa gatcagaggg
     4381 gtgaacttcc catccaacgg ccctgtgatg cagaagaaaa cactcggctg ggaggccaac
     4441 accgagatgc tgtaccccgc tgacggcggc ctggaaggca gaagcgacat ggccctgaag
     4501 ctcgtgggcg ggggccacct gatctgcaac ttcaagacca catacagatc caagaaaccc
     4561 gctaagaacc tcaagatgcc cggcgtctac tatgtggacc acagactgga aagaatcaag
     4621 gaggccgaca aagagaccta cgtcgagcag cacgaggtgg ctgtggccag atactgcgac
     4681 ctccctagca aactggggca caaacttaat tgaaaacgtt gttgtttggg gttgaattac
     4741 tcttctagac cgcggcgcgc ccgataatca acctctggat tacaaaattt gtgaaagatt
     4801 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     4861 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     4921 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     4981 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     5041 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     5101 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     5161 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     5221 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     5281 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     5341 ggccgcctcc ccgcatcgat accgtcgacc tcgatcgaga cctagaaaaa catggagcaa
     5401 tcacaagtag caatacagca gctaccaatg ctgattgtgc ctggctagaa gcacaagagg
     5461 aggaggaggt gggttttcca gtcacacctc aggtaccttt aagaccaatg acttacaagg
     5521 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     5581 cccaacgaag acaagatatc cttgatctgt ggatctacca cacacaaggc tacttccctg
     5641 attggcagaa ctacacacca gggccaggga tcagatatcc actgaccttt ggatggtgct
     5701 acaagctagt accagttgag caagagaagg tagaagaagc caatgaagga gagaacaccc
     5761 gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa gtattagagt
     5821 ggaggtttga cagccgccta gcatttcatc acatggcccg agagctgcat ccggactgta
     5881 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     5941 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     6001 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     6061 gcagcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6121 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6181 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6241 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6301 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6361 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6421 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6481 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6541 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6601 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6661 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6721 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6781 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6841 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6901 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6961 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7021 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7081 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7141 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7201 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7261 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7321 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7381 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7441 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7501 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7561 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7621 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7681 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7741 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7801 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7861 gaaaagtgcc acctgac
//
