LOCUS       ATP1A1_G4_Q118R_        2738 bp    DNA     circular UNA 08-JUN-2022
DEFINITION  Ligation of 2 sequences.
ACCESSION   urn.local...18-ewzuycl
VERSION     urn.local...18-ewzuycl
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     terminator      1..7
                     /standard_name="U6 Terminator"
     promoter        72..312
                     /note="RNA polymerase III promoter for human U6 snRNA"
                     /standard_name="U6 promoter"
     primer_bind     72..92
                     /note="Human U6 promoter, forward primer"
                     /standard_name="hU6-F"
     primer_bind     243..262
                     /note="Human U6 promoter, forward primer"
                     /standard_name="LKO.1 5'"
     misc_RNA        321..340
                     /standard_name="EBFP guide"
     misc_RNA        341..416
                     /standard_name="sgRNA scaffold"
     misc_RNA        417..441
                     /standard_name="EBFP to EGFP extension"
     misc_RNA        442..478
                     /standard_name="tevopreQ1 structural motif"
     terminator      479..485
                     /standard_name="U6 terminator"
     rep_origin      complement(582..1170)
     primer_bind     complement(662..681)
                     /note="pBR322 origin, forward primer"
                     /standard_name="pBR322ori-F"
     CDS             complement(1341..2201)
                     /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     1964..1983
                     /note="Ampicillin resistance gene, reverse primer"
                     /standard_name="Amp-R"
     promoter        complement(2202..2306)
                     /gene="bla"
                     /standard_name="AmpR promoter"
     promoter        2369..2609
                     /note="RNA polymerase III promoter for human U6 snRNA"
                     /standard_name="U6 promoter"
     primer_bind     2369..2389
                     /note="Human U6 promoter, forward primer"
                     /standard_name="hU6-F"
     primer_bind     2540..2559
                     /note="Human U6 promoter, forward primer"
                     /standard_name="LKO.1 5'"
     misc_RNA        2618..2637
                     /standard_name="ATP1A1-G4"
     misc_RNA        2638..2713
                     /standard_name="sgRNA scaffold"
     misc_RNA        2714..2725
                     /standard_name="RT template (Q118R)"
     misc_difference 2721..2723
                     /standard_name="Q118R (CAA to AGA)"
     misc_RNA        2726..2738
                     /standard_name="PBS"
ORIGIN      
        1 tttttttaag cttgggccgc tcgaggtacc tctctacgtc gactggatcc ggtaccaagg
       61 tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata cgatacaagg
      121 ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta gtacaaaata
      181 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa
      241 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct
      301 tgtggaaagg acgaaacacc ggctgaagca ctgcacgccg gttttagagc tagaaatagc
      361 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgcccct
      421 gacctacggc gtgcagtgct tcgcggttct atctagttac gcgttaaacc aactagaatt
      481 tttttaagct tgggccgctc gaggtacctc tctacatatg acatgtgagc aaaaggccag
      541 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
      601 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      661 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      721 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      781 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      841 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      901 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      961 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     1021 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     1081 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     1141 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     1201 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     1261 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     1321 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     1381 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     1441 gagggcttac catctggccc cagtgctgca atgataccgc gagatccacg ctcaccggct
     1501 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     1561 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     1621 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     1681 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     1741 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     1801 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1861 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     1921 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     1981 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     2041 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     2101 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     2161 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     2221 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     2281 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgct
     2341 agctgtacat accaaggtcg ggcaggaaga gggcctattt cccatgattc cttcatattt
     2401 gcatatacga tacaaggctg ttagagagat aattagaatt aatttgactg taaacacaaa
     2461 gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt
     2521 aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt
     2581 cttggcttta tatatcttgt ggaaaggacg aaacaccgtt cctcttctgt agcagctgtt
     2641 ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc
     2701 accgagtcgg tgctagcatc agagctgcta cagaagag
//
