LOCUS       Exported                2335 bp ds-DNA     circular SYN 18-JAN-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    pU6-Sp-gRNA-AAVS1_A3786c_attP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2335)
  AUTHORS   AVA
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jan 18, 2022 from SnapGene 5.0.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..2335
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     24..46
                     /label=AVA037
                     /note="color: black; sequence: GCAGGCTTTAAAGGAACCAATTC"
     promoter        82..330
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     189..208
                     /label=KZ396
                     /note="color: #a020f0; direction: RIGHT"
     misc_feature    331..350
                     /label=spacer
                     /note="color: #3366ff"
     misc_RNA        351..426
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
                     /note="color: #00ccff; direction: RIGHT"
     misc_feature    427..469
                     /label=attP
                     /note="color: #ccffcc"
     misc_feature    470..482
                     /label=PBS
                     /note="color: #339966"
     primer_bind     complement(511..534)
                     /label=AVA038
                     /note="color: black; sequence: GCTCACATGTCATATGTAGAGAGG"
     rep_origin      complement(586..1174)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     misc_feature    complement(749..768)
                     /label=KZ397
                     /note="color: #a6acb3; direction: LEFT"
     primer_bind     1249..1268
                     /label=ACK562
                     /note="color: #a020f0; direction: RIGHT"
     CDS             complement(1345..2205)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 1345 .. 2136 / #ccffcc
                        2: 2137 .. 2205 / #ccffcc / signal sequence
                     Cleavage site after base 2136"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1751..1770
                     /label=ACK563
                     /note="color: #a020f0; direction: RIGHT"
     promoter        complement(2206..2310)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     promoter        complement(2247..2275)
                     /label=amp prom
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 gacgtcgcta gctgtacaaa aaagcaggct ttaaaggaac caattcagtc gactggatcc
       61 ggtaccaagg tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata
      121 cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta
      181 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta
      241 tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct
      301 ttatatatct tgtggaaagg acgaaacacc gctggccccc caccgcccca gttttagagc
      361 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
      421 cggtgctacc gtacaccact gagaccgcgg tggttgacca gacaaacctg gcggtggggg
      481 gcttttttta agcttgggcc gctcgaggta cctctctaca tatgacatgt gagcaaaagg
      541 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
      601 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
      661 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
      721 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
      781 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
      841 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
      901 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
      961 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     1021 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     1081 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     1141 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     1201 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     1261 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     1321 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     1381 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     1441 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     1501 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     1561 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     1621 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     1681 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     1741 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     1801 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     1861 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     1921 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     1981 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     2041 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     2101 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     2161 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     2221 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     2281 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacct
//
