LOCUS       ATP1A1_G6_T804N_        2702 bp    DNA     circular UNA 19-OCT-2021
DEFINITION  Ligation of 2 sequences.
ACCESSION   urn.local...7a-dzvvfvp
VERSION     urn.local...7a-dzvvfvp
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..270
                     /standard_name="MTOR I2017T guide"
     misc_RNA        271..346
                     /standard_name="sgRNA scaffold"
     misc_RNA        347..371
                     /standard_name="MTOR I2017T extension"
     terminator      372..378
                     /standard_name="U6 Terminator"
     rep_origin      complement(475..1063)
     primer_bind     complement(555..574)
                     /note="pBR322 origin, forward primer"
                     /standard_name="pBR322ori-F"
     CDS             complement(1234..2094)
                     /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     1857..1876
                     /note="Ampicillin resistance gene, reverse primer"
                     /standard_name="Amp-R"
     promoter        complement(2095..2199)
                     /gene="bla"
                     /standard_name="AmpR promoter"
     promoter        2262..2502
                     /note="RNA polymerase III promoter for human U6 snRNA"
                     /standard_name="U6 promoter"
     primer_bind     2262..2282
                     /note="Human U6 promoter, forward primer"
                     /standard_name="hU6-F"
     primer_bind     2433..2452
                     /note="Human U6 promoter, forward primer"
                     /standard_name="LKO.1 5'"
     misc_RNA        2511..2530
                     /standard_name="ATP1A1-G6"
     misc_RNA        2531..2606
                     /standard_name="sgRNA scaffold"
     misc_RNA        2607..2631
                     /standard_name="ATP1A1 T804N extension"
     misc_RNA        2607..2619
                     /standard_name="RT remplate"
     misc_difference complement(2610..2612)
                     /standard_name="T804N (ACT to AAT)"
     misc_difference 2619
                     /standard_name="G6 seed mutation"
     misc_RNA        2620..2631
                     /standard_name="PBS"
     terminator      2632..2638
                     /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 catgccagag gatggccact gttttagagc tagaaatagc aagttaaaat
      301 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgcgctg acacgagtgg
      361 ccatcctctg gtttttttaa gcttgggccg ctcgaggtac ctctctacat atgacatgtg
      421 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
      481 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
      541 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
      601 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
      661 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
      721 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
      781 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
      841 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
      901 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
      961 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     1021 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     1081 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     1141 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     1201 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     1261 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     1321 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagatcc
     1381 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     1441 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     1501 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     1561 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     1621 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     1681 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     1741 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     1801 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     1861 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     1921 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     1981 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     2041 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     2101 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     2161 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     2221 acctgacgtc gctagctgta cataccaagg tcgggcagga agagggccta tttcccatga
     2281 ttccttcata tttgcatata cgatacaagg ctgttagaga gataattaga attaatttga
     2341 ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt
     2401 agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa
     2461 agtatttcga tttcttggct ttatatatct tgtggaaagg acgaaacacc gcaaacattc
     2521 cactaccact gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac
     2581 ttgaaaaagt ggcaccgagt cggtgcgaca ttccccagag gtagtggaat gtttttttaa
     2641 gcttgggccg ctcgaggtac ctctctacgt cgactggatc cggtaccaag gtcgggcagg
     2701 aa
//
