LOCUS       sequence-436395-        3128 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid codes for the guide of the OgeuIscB with a CAG target.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3128)
  AUTHORS   Mangin A, Dion V, Menzies G
  TITLE     Developing small Cas9 hybrids using molecular modeling
  JOURNAL   bioRxiv 2023.10.24.563270
REFERENCE   2  (bases 1 to 3128)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2023.10.24.563270"
COMMENT     Sequence Label: OgeuIscB CAG RNA
FEATURES             Location/Qualifiers
     source          1..3128
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     265..284
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     518..535
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    652..673
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(688..693,694..711,712..718)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    726..742
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     731..753
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     750..766
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     750..766
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        780..1028
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     780..800
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     951..970
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(1278..1295)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1278..1294)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1287..1309)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1503..1522)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1622..1644
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1682..1700)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1768..1872
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1873..1941,1942..2733)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2091..2110)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(2904..3128,1..364)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
       61 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
      121 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
      181 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
      241 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
      301 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
      361 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
      421 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
      481 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      541 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
      601 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
      661 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
      721 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccag
      781 agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga
      841 taattggaat taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa
      901 agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata
      961 tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac
     1021 gaaacaccgc agcagcagca gcagcagggc tcttccaact ttatggttgc gaccgtaggt
     1081 tgaaagagca caggctgaga cattcgtaag gccgaaagac cggacgcacc ctgggatttc
     1141 cccagtcccc ggaactgcat agcggatgcc agttgatgga gcaatctatc agataagcca
     1201 gggggaacaa tcacctctct gtatcagaga gagttttaca aaaggaggaa cggttttttg
     1261 gtaccgagct cgaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     1321 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     1381 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg
     1441 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
     1501 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     1561 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     1621 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     1681 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     1741 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     1801 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     1861 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     1921 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     1981 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     2041 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     2101 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     2161 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     2221 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     2281 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     2341 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     2401 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     2461 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     2521 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     2581 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     2641 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     2701 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     2761 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     2821 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     2881 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     2941 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3001 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     3061 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3121 ctaatcct
//
