LOCUS       pDY2259_hNOLC1_atgRNA_P 2359 bp ds-DNA     circular     08-MAY-2024
DEFINITION  .
KEYWORDS    "creator:MIT" "marker:AmpR, Amp-R, SPACER"
FEATURES             Location/Qualifiers
     primer_bind     393..412
                     /label="Amp-R"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /note="label: Amp-R"
     promoter        complement(631..735)
                     /label="AmpR promoter"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /note="label: AmpR promoter"
     primer_bind     842..862
                     /label="hU6-F"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
                     /note="label: hU6-F"
     promoter        842..1090
                     /label="U6 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="note: RNA polymerase III promoter for human U6 snRNA (Domitrovich & Kunkel, 2003)"
     primer_bind     1013..1032
                     /label="LKO.1 5'"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
                     /note="label: LKO.1 5'"
     misc_feature    1091..1110
                     /label="SPACER"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_RNA        1111..1186
                     /label="gRNA scaffold"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
                     /note="note: guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system"
     protein_bind    1191..1228
                     /label="Bxb1 attB"
                     /ApEinfo_revcolor="#31849b"
                     /ApEinfo_fwdcolor="#31849b"
                     /note="bound_moiety: Mycobacteriophage Bxb1 integrase note: Minimal attB site for the Mycopbacteriophage Bxb1 integrase (Voutev and Mann, 2017)"
     rep_origin      complement(1345..1958)
                     /label="ori"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
                     /note="direction: LEFT label: ori"
     primer_bind     complement(1450..1469)
                     /label="pBR322ori-F"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
                     /note="label: pBR322ori-F"
     CDS             complement(2129..630)
                     /label="AmpR"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
                     /note="label: AmpR"
ORIGIN
        1 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
       61 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
      121 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
      181 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
      241 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
      301 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
      361 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
      421 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
      481 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
      541 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
      601 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
      661 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
      721 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgcta gctgtacaaa
      781 aaagcaggct ttaaaggaac caattcagtc gactggatcc ggtaccaagg tcgggcagga
      841 agagggccta tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga
      901 gataattaga attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag
      961 aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca
     1021 tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg
     1081 acgaaacacc GCGTATTGCC TGGAGGATGG gttttagagc tagaaatagc aagttaaaat
     1141 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgcccgg atgatcctga
     1201 cgacggagac cgccgtcgtc gacaagccgg cctcctccag gtttttttaa gcttgggccg
     1261 ctcgaggtac ctctctacat atgacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     1321 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1381 aaatcgacgc tcaaggtttt agagctaggc caacatgagg tcagaggtgg cgaaacccga
     1441 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1501 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1561 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1621 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1681 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     1741 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     1801 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1861 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     1921 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1981 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     2041 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2101 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     2161 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     2221 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gatccacgct
     2281 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     2341 gtcctgcaac tttatccgc
//