LOCUS       194694                 10082 bp    DNA     circular UNA 26-JUN-2023
DEFINITION  pLKO.1_U6nt-shRNA_CMV_NES-CaProLa_pos-ctrl-meGFP.
ACCESSION   urn.local...2j-gfr95wy
VERSION     urn.local...2j-gfr95wy
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     exon            289..1128
                     /standard_name="Puromycin Resisteance cassette"
     gene            1256..1308
                     /note="Geneious type: Gene"
                     /standard_name="delta_U3"
     gene            1309..1489
                     /note="Geneious type: Gene"
                     /standard_name="HIV-1_5_LTR; truncHIV-1_3_LTR"
     misc_feature    1722..1799
                     /note="Geneious type: Origin of Replication"
                     /standard_name="SV40 origin"
     promoter        complement(1878..1896)
                     /note="Geneious type: Promoter"
                     /standard_name="T7 prom"
     misc_feature    complement(1879..1896)
                     /gene="T7 promoter"
                     /standard_name="T7 promoter misc"
     misc_feature    1903..2063
                     /note="Geneious type: Reporter"
                     /standard_name="lacZ_a reporter"
     misc_feature    complement(2064..2519)
                     /note="Geneious type: motif"
                     /standard_name="F1 Ori"
     promoter        2580..2608
                     /note="Geneious type: Promoter"
                     /standard_name="amp prom"
     CDS             2650..3510
                     /standard_name="AmpR"
     misc_feature    3665..4284
                     /note="Geneious type: Origin of Replication"
                     /standard_name="pBR322 origin"
     promoter        4593..4622
                     /note="Geneious type: Promoter"
                     /standard_name="lac prom"
     promoter        4692..4711
                     /note="Geneious type: Promoter"
                     /standard_name="T3 prom"
     gene            4965..5145
                     /note="Geneious type: Gene"
                     /standard_name="HIV-1_5_LTR; truncHIV-1_3_LTR"
     gene            5256..5300
                     /note="Geneious type: Gene"
                     /standard_name="HIV-1_psi_pack"
     misc_feature    5811..6044
                     /note="Geneious type: Regulatory Sequence"
                     /standard_name="RRE reg"
     promoter        6557..6813
                     /note="Geneious type: Promoter"
                     /standard_name="hU6 prom"
     ncRNA           6814..7341
                     /standard_name="non-targeted shRNA"
     promoter        7342..7492
                     /standard_name="CMV"
     primer_bind     <7493..7512
                     /standard_name="jh3194"
     CDS             7493..9868
                     /standard_name="CaProLa_Pos_ctrl"
     CDS             7493..7555
                     /standard_name="NES (CaMPARI)"
     misc_feature    7556..7558
                     /note="Geneious type: motif"
                     /standard_name="A145"
     CDS             <7556..>7585
                     /standard_name="HaloPep145-154"
     misc_feature    7580..7582
                     /note="Geneious type: motif"
                     /standard_name="R153"
     misc_feature    7583..7585
                     /note="Geneious type: motif"
                     /standard_name="T154"
     misc_feature    7586..7591
                     /note="Geneious type: motif"
                     /standard_name="GS"
     CDS             7592..8032
                     /codon_start=1
                     /product="calcium-binding protein"
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTE
                     AELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAA
                     ELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVVMMTAK"
                     /standard_name="calcium-binding protein CDS"
     CDS             8039..8128
                     /standard_name="Pro30"
     CDS             8138..8215
                     /standard_name="M13_2.07"
     misc_feature    8216..8224
                     /note="Geneious type: motif"
                     /standard_name="GGS"
     misc_feature    8225..8227
                     /note="Geneious type: motif"
                     /standard_name="F144"
     CDS             <8225..>8680
                     /note="Geneious type: cds"
                     /standard_name="HaloTag7"
     misc_feature    8681..8725
                     /note="Geneious type: motif"
                     /standard_name="(GGS/T)x5"
     CDS             <8726..>9139
                     /note="Geneious type: cds"
                     /standard_name="HaloTag7"
     misc_feature    9137..9139
                     /note="Geneious type: motif"
                     /standard_name="Trp141"
     CDS             <9140..9148
                     /codon_start=1
                     /standard_name="GGS-Linker"
     CDS             9149..9865
                     /note="Geneious type: cds"
                     /standard_name="eGFP"
     primer_bind     complement(9845..9888)
                     /standard_name="jh3195"
     terminator      9866..9868
                     /standard_name="TAA-STOP"
     exon            9869..9997
                     /standard_name="cPPT"
     primer_bind     9869..9897
                     /standard_name="jh3192"
     exon            9998..288
                     /standard_name="hPGK 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 gatccaccgg agcttaccat gaccgagtac
      541 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
      601 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
      661 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
      721 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
      781 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
      841 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
      901 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
      961 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     1021 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     1081 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg cccgccccac
     1141 gacccgcagc gcccgaccga aaggagcgca cgaccccatg catcggtacc tttaagacca
     1201 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
     1261 gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg
     1321 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     1381 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     1441 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca
     1501 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
     1561 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     1621 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     1681 tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc ctaactccgc
     1741 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     1801 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     1861 ggacgtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt
     1921 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     1981 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     2041 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2101 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     2161 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2221 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     2281 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     2341 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     2401 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     2461 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     2521 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     2581 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     2641 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     2701 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     2761 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     2821 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     2881 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     2941 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     3001 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     3061 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     3121 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     3181 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     3241 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     3301 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     3361 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     3421 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     3481 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     3541 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     3601 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     3661 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3721 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3781 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     3841 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3901 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3961 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4021 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4081 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4141 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4201 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4261 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4321 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     4381 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     4441 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     4501 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     4561 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     4621 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     4681 gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaatgt
     4741 agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct
     4801 tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg
     4861 ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc
     4921 attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag
     4981 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     5041 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     5101 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag
     5161 ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga
     5221 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag
     5281 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag
     5341 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca
     5401 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac
     5461 atcacgaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     5521 aagaacttac atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     5581 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     5641 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
     5701 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     5761 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     5821 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     5881 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     5941 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     6001 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     6061 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     6121 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     6181 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     6241 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     6301 ctgtggtata taaaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt
     6361 ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca
     6421 gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg
     6481 agagagagac agagacagat ccattcgatt agtgaacgga tctcgacggt atcgatcacg
     6541 agactagcct cgagcggccg cccccttcac cgagggccta tttcccatga ttccttcata
     6601 tttgcatata cgatacaagg ctgttagaga gataattgga attaatttga ctgtaaacac
     6661 aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt
     6721 tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga
     6781 tttcttggct ttatatatct tgtggaaagg acgaaacacc gggcgcgata gcgctaataa
     6841 tttctcgaga aattattagc gctatcgcgc tttttgaatt cagttattaa tagtaatcaa
     6901 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     6961 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     7021 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     7081 aagctgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     7141 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     7201 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     7261 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     7321 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     7381 agaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     7441 gcagagctgg tttagtgaac cgtcagatcc gctagcgctc ccggtcgcca ccatgctgca
     7501 gaacgagctt gctcttaagt tggctggact tgatattaac aagactggag gttctgcccg
     7561 cgagaccttc caggccttcc gcaccggttc tgaccaactg actgaagagc agatcgcaga
     7621 atttaaagag gctttctccc tatttgacaa ggacggggat gggacaataa caaccaagga
     7681 gctggggacg gtgatgcggt ctctggggca gaaccccaca gaagcagagc tgcaggacat
     7741 gatcaatgaa gtagatgccg acggtgacgg cacaatcgac ttccctgagt tcctgacaat
     7801 gatggcaaga aaaatgaaag acacagacag tgaagaagaa attagagaag cgttccgtgt
     7861 gtttgataag gatggcaatg gctacatcag tgcagcagag cttcgccacg tgatgacaaa
     7921 ccttggagag aagttaacag atgaagaggt tgatgaaatg atcagggaag cagacatcga
     7981 tggggatggt caggtaaact acgaagagtt tgtagtgatg atgacagcga aggaattccc
     8041 tcctcctccg ccaccgccac caccgccgcc gccgccacca ccacctcctc cgcctcctcc
     8101 accaccaccg ccaccgcctc caccacctgg cggcagcagg gtcgactcat cacgtcgtaa
     8161 gttcaataag acaggtcacg cactgagagc tataggtcgg ctgagctcac tcgagggcgg
     8221 ttcttttgcc cgcgagacct tccaggcctt ccgcaccacc gacgtcggcc gcaagctgat
     8281 catcgatcag aacgttttta tcgagggtac gctgccgatg ggtgtcgtcc gcccgctgac
     8341 tgaagtcgag atggaccatt accgcgagcc gttcctgaat cctgttgacc gcgagccact
     8401 gtggcgcttc ccaaacgagc tgccaatcgc cggtgagcca gcgaacatcg tcgcgctggt
     8461 cgaagaatac atggactggc tgcaccagtc ccctgtcccg aagctgctgt tctggggcac
     8521 cccaggcgtt ctgatcccac cggccgaagc cgctcgcctg gccaaaagcc tgcctaactg
     8581 caaggctgtg gacatcggcc cgggtctgaa tctgctgcaa gaagacaacc cggacctgat
     8641 cggcagcgag atcgcgcgct ggctgtctac tctggagatt ggtggcaccg gtggctctgg
     8701 cggtaccggt ggctctggcg gttctatcgg tactggcttt ccattcgacc cccattatgt
     8761 ggaagtcctg ggcgagcgca tgcactacgt cgatgttggt ccgcgcgatg gcacccctgt
     8821 gctgttcctg cacggtaacc cgacctcctc ctacgtgtgg cgcaacatca tcccgcatgt
     8881 tgcaccgacc catcgctgca ttgctccaga cctgatcggt atgggcaaat ccgacaaacc
     8941 agacctgggt tatttcttcg acgaccacgt ccgcttcatg gatgccttca tcgaagccct
     9001 gggtctggaa gaggtcgtcc tggtcattca cgactggggc tccgctctgg gtttccactg
     9061 ggccaagcgc aatccagagc gcgtcaaagg tattgcattt atggagttca tccgccctat
     9121 cccgacctgg gacgaatggg gaggttctat ggtgagcaag ggcgaggagc tgttcaccgg
     9181 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     9241 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     9301 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     9361 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     9421 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     9481 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     9541 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     9601 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     9661 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     9721 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccaaac tgagcaaaga
     9781 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     9841 tctcggcatg gacgagctgt acaagtaaaa ttaattctcg acctcgagac aaatggcagt
     9901 attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat
     9961 agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat
    10021 tcaaaatttt cgggtttatt acagggacag cagagatcca ctttggccgc ggctcgaggg
    10081 gg
//
