LOCUS       AAV-U6-sgPlxnb2-KO-4-EF 5593 bp ds-DNA     circular     15-JUN-2022
DEFINITION  .
FEATURES             Location/Qualifiers
     repeat_region   complement(1..141)
                     /label="AAV2 ITR"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
                     /note="note: pLannotate database: snapgene identity: 100.0 match_length: 100.0 fragment: False other: repeat_region"
     primer          140..177
                     /label="InF-U6-F"
                     /note="sequence: ctgcggcccccgttagagggcctatttcccatgattcc"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     promoter        155..395
                     /label="U6 promoter"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
                     /note="note: pLannotate database: snapgene identity: 100.0 match_length: 100.0 fragment: False other: promoter"
     misc_feature    193..1316
                     /label="sanger (hU6-f)"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          400..424
                     /label="AAV-sgPlxnb2-4-EF1a-LibVec FWD"
                     /note="sequence: caccGACCGTCCATCCGTATCAGCA"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     primer          complement(404..428)
                     /label="AAV-sgPlxnb2-4-EF1a-LibVec REV"
                     /note="sequence: aaacTGCTGATACGGATGGACGGTC"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     misc_feature    405..424
                     /label="Guide"
                     /ApEinfo_revcolor="#7ea7d8"
                     /ApEinfo_fwdcolor="#7ea7d8"
     misc_RNA        425..500
                     /label="gRNA scaffold"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
                     /note="note: pLannotate database: snapgene identity: 100.0 match_length: 100.0 fragment: False other: misc_RNA"
     primer          475..493
                     /label="qPCR-gRNA-F"
                     /note="sequence: ttgaaaaagtggcaccgag"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     primer          complement(486..521)
                     /label="InF-gRNA-R"
                     /note="sequence: tccatctttgcattaaaaaaagcaccgactcggtgc"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     CDS             517..583
                     /label="small t antigen (fragment)"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /note="note: pLannotate database: snapgene identity: 100.0 match_length: 12.8 fragment: True other: CDS"
     CDS             517..583
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="MDKVLNREESLQLMDLLGLERS"
     primer          complement(568..587)
                     /label="qPCR-EF1a-R"
                     /note="sequence: ttcccactcctttcaagacc"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     promoter        590..1768
                     /label="EF-1α promoter"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
                     /note="note: pLannotate database: snapgene identity: 100.0 match_length: 100.0 fragment: False other: promoter"
     CDS             1782..2501
                     /label="EGFP"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
                     /note="note: pLannotate database: snapgene identity: 100.0 match_length: 100.0 fragment: False other: CDS"
     CDS             1782..2501
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK*"
     promoter        complement(2569..2587)
                     /label="SP6 promoter"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /note="note: pLannotate database: snapgene identity: 100.0 match_length: 100.0 fragment: False other: promoter"
     polyA_signal    2613..2837
                     /label="bGH poly(A) signal"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
                     /note="note: pLannotate database: snapgene identity: 100.0 match_length: 100.0 fragment: False other: polyA_signal"
     repeat_region   2856..2996
                     /label="AAV2 ITR"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
                     /note="note: pLannotate database: snapgene identity: 100.0 match_length: 100.0 fragment: False other: repeat_region"
     misc_feature    complement(3001..3052)
                     /label="bom (fragment)"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /note="note: pLannotate database: snapgene identity: 100.0 match_length: 36.9 fragment: True other: misc_feature"
     rep_origin      3071..3451
                     /label="M13 ori"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /note="note: pLannotate database: snapgene identity: 100.0 match_length: 100.0 fragment: False other: rep_origin"
     misc_feature    complement(3527..3576)
                     /label="bom (fragment)"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
                     /note="note: pLannotate database: snapgene identity: 100.0 match_length: 35.5 fragment: True other: misc_feature"
     CDS             complement(3678..3712)
                     /label="rop (fragment)"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: pLannotate database: snapgene identity: 100.0 match_length: 18.2 fragment: True other: CDS"
     CDS             complement(3678..3712)
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="SRVSVMTVKTS"
     promoter        3808..3912
                     /label="AmpR promoter"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
                     /note="note: pLannotate database: snapgene identity: 100.0 match_length: 100.0 fragment: False other: promoter"
     CDS             3913..4773
                     /label="AmpR"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
                     /note="note: pLannotate database: snapgene identity: 99.8 match_length: 100.0 fragment: False other: CDS"
     CDS             3913..4773
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*"
     rep_origin      4944..5532
                     /label="ori"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
                     /note="note: pLannotate database: snapgene identity: 100.0 match_length: 100.0 fragment: False other: rep_origin"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccccc gttagagggc ctatttccca tgattccttc
      181 atatttgcat atacgataca aggctgttag agagataatt agaattaatt tgactgtaaa
      241 cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc
      301 agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt
      361 cgatttcttg gctttatata tcttgtggaa aggacgaaac accGACCGTC CATCCGTATC
      421 AGCAgtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa
      481 aagtggcacc gagtcggtgc tttttttaat gcaaagatgg ataaagtttt aaacagagag
      541 gaatctttgc agctaatgga ccttctaggt cttgaaagga gtgggaattg gctccggtgc
      601 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
      661 caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
      721 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
      781 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc
      841 ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg
      901 gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag
      961 gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg
     1021 gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc
     1081 tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct
     1141 tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga
     1201 cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc
     1261 gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc
     1321 gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc
     1381 ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc
     1441 gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt
     1501 cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag
     1561 cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca
     1621 cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga
     1681 atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag
     1741 tttttttctt ccatttcagg tgtcgtgagc ggccgctcga gatggtgagc aagggcgagg
     1801 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     1861 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     1921 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     1981 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     2041 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     2101 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     2161 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     2221 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     2281 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     2341 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     2401 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     2461 ccgccgggat cactctcggc atggacgagc tgtacaagta agagcgcacc cgtccgagct
     2521 aaggcgcgcc atacatgcat ggtgtacaag taatctagag ggccctattc tatagtgtca
     2581 cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     2641 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2701 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2761 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     2821 tgcggtgggc tctatggctt ctgaggcggg gccgcaggaa cccctagtga tggagttggc
     2881 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     2941 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     3001 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     3061 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3121 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3181 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     3241 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     3301 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3361 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc gggctattct
     3421 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3481 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     3541 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     3601 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccttc
     3661 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gatacgaaag
     3721 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     3781 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3841 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3901 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     3961 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4021 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4081 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     4141 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     4201 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     4261 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     4321 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     4381 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     4441 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     4501 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     4561 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     4621 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     4681 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     4741 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     4801 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4861 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4921 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4981 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5041 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     5101 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5161 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5221 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5281 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5341 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5401 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5461 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5521 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5581 tttgctcaca tgt
//