LOCUS       ATP1A1_G6_T804N_        2701 bp    DNA     circular UNA 19-OCT-2021
DEFINITION  Ligation of 2 sequences.
ACCESSION   urn.local...7i-dzvvg0i
VERSION     urn.local...7i-dzvvg0i
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     promoter        1..241
                     /note="RNA polymerase III promoter for human U6 snRNA"
                     /standard_name="U6 promoter"
     primer_bind     1..21
                     /note="Human U6 promoter, forward primer"
                     /standard_name="hU6-F"
     primer_bind     172..191
                     /note="Human U6 promoter, forward primer"
                     /standard_name="LKO.1 5'"
     misc_RNA        250..269
                     /standard_name="MTOR F2108L guide"
     misc_RNA        270..345
                     /standard_name="sgRNA scaffold"
     misc_RNA        346..370
                     /standard_name="MTOR F2108L extension"
     terminator      371..377
                     /standard_name="U6 Terminator"
     rep_origin      complement(474..1062)
     primer_bind     complement(554..573)
                     /note="pBR322 origin, forward primer"
                     /standard_name="pBR322ori-F"
     CDS             complement(1233..2093)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
                     /standard_name="AmpR"
     primer_bind     1856..1875
                     /note="Ampicillin resistance gene, reverse primer"
                     /standard_name="Amp-R"
     promoter        complement(2094..2198)
                     /gene="bla"
                     /standard_name="AmpR promoter"
     promoter        2261..2501
                     /note="RNA polymerase III promoter for human U6 snRNA"
                     /standard_name="U6 promoter"
     primer_bind     2261..2281
                     /note="Human U6 promoter, forward primer"
                     /standard_name="hU6-F"
     primer_bind     2432..2451
                     /note="Human U6 promoter, forward primer"
                     /standard_name="LKO.1 5'"
     misc_RNA        2510..2529
                     /standard_name="ATP1A1-G6"
     misc_RNA        2530..2605
                     /standard_name="sgRNA scaffold"
     misc_RNA        2606..2630
                     /standard_name="ATP1A1 T804N extension"
     misc_RNA        2606..2618
                     /standard_name="RT remplate"
     misc_difference complement(2609..2611)
                     /standard_name="T804N (ACT to AAT)"
     misc_difference 2618
                     /standard_name="G6 seed mutation"
     misc_RNA        2619..2630
                     /standard_name="PBS"
     terminator      2631..2637
                     /standard_name="U6 terminator"
ORIGIN      
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg cagctgcttt gagattcgtg ttttagagct agaaatagca agttaaaata
      301 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgctgtgt tgcgacgaat
      361 ctcaaagcag tttttttaag cttgggccgc tcgaggtacc tctctacata tgacatgtga
      421 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      481 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      541 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      601 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      661 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      721 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      781 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      841 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      901 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      961 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     1021 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     1081 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     1141 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     1201 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1261 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1321 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagatcca
     1381 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1441 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1501 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1561 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1621 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1681 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1741 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1801 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1861 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1921 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1981 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     2041 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     2101 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     2161 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     2221 cctgacgtcg ctagctgtac ataccaaggt cgggcaggaa gagggcctat ttcccatgat
     2281 tccttcatat ttgcatatac gatacaaggc tgttagagag ataattagaa ttaatttgac
     2341 tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta
     2401 gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa
     2461 gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg caaacattcc
     2521 actaccactg ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact
     2581 tgaaaaagtg gcaccgagtc ggtgcgacat tccccagagg tagtggaatg tttttttaag
     2641 cttgggccgc tcgaggtacc tctctacgtc gactggatcc ggtaccaagg tcgggcagga
     2701 a
//
