LOCUS       pAAV_KI;Sptbn1_Donor;smFP-V5_        5049 bp    DNA     circular UNA 11-JUL-2025
DEFINITION  pAAV-U6sgRNA(SapI)_hSyn-GFP-KASH-bGH (SpGuide acceptor). AAV
            plasmid for sgRNA cloning. GFP-KASH fusion facilitates FACS sorting
            of cells and nuclei. .
ACCESSION   urn.local...14-jebled8
VERSION     urn.local...14-jebled8
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     repeat_region   1..141
                     /locus_tag="AAV2 ITR"
                     /note="inverted terminal repeat of adeno-associated virus
                     serotype 2"
                     /standard_name="AAV2 ITR"
     repeat_region   12..141
                     /locus_tag="AAV2 ITR(1)"
                     /standard_name="AAV2 ITR"
     misc_feature    158..398
                     /standard_name="hU6"
     misc_feature    408..427
                     /standard_name="mouse Sptbn1 Cterm"
     misc_feature    428..513
                     /standard_name="Scaffold cr1"
     misc_feature    520..833
                     /standard_name="mU6"
     protein_bind    714..747
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (GCATACAT)."
                     /standard_name="loxP"
     primer_bind     751..771
                     /standard_name="mU6-F"
     misc_feature    843..862
                     /locus_tag="DRSR2(1)"
                     /standard_name="DRSR2"
     misc_feature    863..950
                     /standard_name="Scaffold cr2"
     misc_feature    963..985
                     /locus_tag="DRSR2(2)"
                     /standard_name="DRSR2"
     CDS             991..1032
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /standard_name="V5 tag"
     misc_feature    991..2226
                     /standard_name="smFP-V5"
     CDS             1042..1083
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /standard_name="V5 tag"
     CDS             1093..1134
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /standard_name="V5 tag"
     misc_feature    1537..1542
                     /standard_name="gaagac>gaggat, mutation on BbsI site"
     CDS             1666..1707
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /standard_name="V5 tag"
     CDS             1717..1758
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /standard_name="V5 tag"
     CDS             1777..1818
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /standard_name="V5 tag"
     CDS             1828..1869
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /standard_name="V5 tag"
     CDS             2083..2124
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /standard_name="V5 tag"
     CDS             2134..2175
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /standard_name="V5 tag"
     CDS             2185..2226
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /standard_name="V5 tag"
     misc_feature    2255..2269
                     /locus_tag="X"
                     /standard_name="X"
     misc_feature    2276..2298
                     /locus_tag="DRSR2(1)"
                     /standard_name="DRSR2"
     repeat_region   2312..2452
                     /locus_tag="AAV2 ITR(2)"
                     /note="inverted terminal repeat of adeno-associated virus
                     serotype 2"
                     /standard_name="AAV2 ITR"
     repeat_region   2312..2441
                     /locus_tag="AAV2 ITR(3)"
                     /standard_name="AAV2 ITR"
     rep_origin      2527..2982
     primer_bind     complement(2614..2633)
                     /locus_tag="F1ori-R"
                     /note="F1 origin, reverse primer"
                     /standard_name="F1ori-R"
     primer_bind     2824..2845
                     /locus_tag="F1ori-F"
                     /note="F1 origin, forward primer"
                     /standard_name="F1ori-F"
     primer_bind     complement(2999..3018)
                     /locus_tag="pRS-marker"
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
                     /standard_name="pRS-marker"
     primer_bind     3118..3140
                     /locus_tag="pGEX 3'"
                     /note="pGEX vectors, reverse primer"
                     /standard_name="pGEX 3'"
     primer_bind     complement(3178..3196)
                     /locus_tag="pBRforEco"
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
                     /standard_name="pBRforEco"
     regulatory      3264..3368
                     /note="Geneious type: promoter"
                     /gene="bla"
                     /locus_tag="AmpR promoter"
                     /standard_name="AmpR promoter"
     CDS             3369..4229
                     /codon_start=1
                     /gene="bla"
                     /locus_tag="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /product="beta-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
                     /standard_name="AmpR"
     primer_bind     complement(3587..3606)
                     /locus_tag="Amp-R"
                     /note="Ampicillin resistance gene, reverse primer"
                     /standard_name="Amp-R"
     rep_origin      4400..4988
     primer_bind     4889..4908
                     /locus_tag="pBR322ori-F"
                     /note="pBR322 origin, forward primer"
                     /standard_name="pBR322ori-F"
ORIGIN      
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgct cacatgtgag ggcctatttc ccatgattcc
      181 ttcatatttg catatacgat acaaggctgt tagagagata attggaatta atttgactgt
      241 aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt
      301 tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta
      361 tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccgctt ctccctcttg
      421 ccaggacgtt taagagctaa gctggaaaca gcatagcaag tttaaataag gctagtccgt
      481 tatcaacttg aaaaagtggc accgagtcgg tgcttttttg atccgacgcg ccatctctag
      541 gcccgcgccg gccccctcgc acggacttgt gggagaagct cggctactcc cctgccccgg
      601 ttaatttgca tataatattt cctagtaact atagaggctt aatgtgcgat aaaagacaga
      661 taatctgttc tttttaatac tagctacatt ttacatgata ggcttggatt tctataactt
      721 cgtatagcat acattatacg aagttataaa cagcacaaaa ggaaactcac cctaactgta
      781 aagtaattgt gtgttttgag actataagta tcccttggag aaccaccttg ttggaaacac
      841 cggcgatcgt aatcacccga gtgtttgaga gctaagcaga aagctgcata gcaagttcaa
      901 ataaggctag tccgtacaca acttgaaaaa gtggcagccg agtcggctgc ttttttctgc
      961 aggcgatcgt aatcacccga gtgggctcga ggaaaaccta taccgaaccc cctccttgga
     1021 cttgatagta ctcagcagca gggtaagccc attccaaacc cacttctcgg tctggatagc
     1081 acacagcaac aaggtaagcc tatccctaac cctctgttgg gcctggactc cacgggccaa
     1141 ggtgtgagta agggcgaaga actgtttacc ggcgtggtcc caattctggt tgagctcgac
     1201 ggtgacgtta acggacacaa gtttagtgtg cgcggagaag gagagggaga tgccaccaac
     1261 ggaaaattga cgctgaagtt tatatgcacc acgggaaagc tgccggtgcc gtggccgacc
     1321 ttggttacaa ccctgggtgg cggagtccag tgtttcagcc gttacccaga ccacatgaag
     1381 cagcatgact tctttaagag tgctatgccg gagggatacg tgcaggaacg gacgatcagt
     1441 ttcaaagatg acggtactta caagacgcga gccgaggtga agttcgaggg agacaccttg
     1501 gtgaacagga tagaacttaa gggaattgat ttcaaggagg atggtaatat tctgggtcac
     1561 aagctggaat acaactttaa cagccataac gtctacatta ctgccgacaa gcagaagaac
     1621 ggcataaagg cgaatttcaa aattcggcac aacgtggaag gtggtggcaa gccgatcccg
     1681 aatcccctgc tgggcttgga tagcacgcag caacagggaa agcccatccc taaccccttg
     1741 ctgggactcg acagcaccgg aggccagcaa ggcggtggca aaccgatacc caatcccctg
     1801 ctcggcctgg atagcaccca gcagcaaggc aagccaatcc ccaaccctct gctgggcctc
     1861 gactcgacag gtggcgatgg aagtgttcaa ctggcggacc actatcagca gaatacgccg
     1921 atcggagatg gccccgtgct gttgccggat aatcactatc tgtcgactca gagcgttctg
     1981 agtaaggacc caaacgaaaa acgagatcac atggtgttgt tggagttcgt gaccgccgcc
     2041 ggcattacgt tgggaatgga cgagctgtac aagggccaag gaggcaagcc cataccaaat
     2101 cccttgctgg gcctcgacag cactcagcaa caaggaaagc caataccaaa tccactgttg
     2161 ggcctcgaca gtacgcaaca gcagggcaag cccatcccga acccgttgct gggactggat
     2221 agtacctaaa ccggtgctcg agaattcgat atcatagcta gctagctagg gatccgcgat
     2281 cgtaatcacc cgagtgggct agaggggccg caggaacccc tagtgatgga gttggccact
     2341 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     2401 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     2461 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     2521 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2581 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     2641 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     2701 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     2761 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     2821 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcgggc tattcttttg
     2881 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     2941 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     3001 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     3061 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     3121 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     3181 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     3241 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3301 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3361 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3421 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3481 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3541 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3601 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3661 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3721 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3781 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3841 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3901 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     3961 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     4021 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     4081 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     4141 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4201 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     4261 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4321 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4381 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4441 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4501 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     4561 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4621 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4681 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4741 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4801 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4861 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4921 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4981 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5041 ctcacatgt
//
