LOCUS       sequence-449444-        3128 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid codes for the guide of the OgeuIscB with a CTG 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 CTG RNA
FEATURES             Location/Qualifiers
     source          1..3128
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(376..1167,1168..1236))
                     /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     999..1018
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1237..1341)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1409..1427
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1465..1487)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1587..1606
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1800..1822
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1814..1831
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1815..1831
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2081..2329)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(2139..2158)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(2309..2329)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(2343..2359)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2343..2359)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2356..2378)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2367..2383
                     /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)."
     promoter        complement(join(2391..2397,2398..2415,2416..2421))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2436..2457
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2574..2591)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(2745..3128,1..205))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2825..2844)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
       61 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
      121 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
      181 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
      241 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
      301 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
      361 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
      421 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
      481 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
      541 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
      601 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
      661 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
      721 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
      781 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
      841 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
      901 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
      961 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     1021 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     1081 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     1141 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     1201 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     1261 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     1321 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     1381 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     1441 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     1501 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     1561 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     1621 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     1681 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     1741 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     1801 ccagtcacga cgttgtaaaa cgacggccag tgaattcgag ctcggtacca aaaaaccgtt
     1861 cctccttttg taaaactctc tctgatacag agaggtgatt gttccccctg gcttatctga
     1921 tagattgctc catcaactgg catccgctat gcagttccgg ggactgggga aatcccaggg
     1981 tgcgtccggt ctttcggcct tacgaatgtc tcagcctgtg ctctttcaac ctacggtcgc
     2041 aaccataaag ttggaagagc ccagcagcag cagcagcagc ggtgtttcgt cctttccaca
     2101 agatatataa agccaagaaa tcgaaatact ttcaagttac ggtaagcata tgatagtcca
     2161 ttttaaaaca taattttaaa actgcaaact acccaagaaa ttattacttt ctacgtcacg
     2221 tattttgtac taatatcttt gtgtttacag tcaaattaat tccaattatc tctctaacag
     2281 ccttgtatcg tatatgcaaa tatgaaggaa tcatgggaaa taggccctct ggcgtaatca
     2341 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     2401 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     2461 gcgttgcgtt cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     2521 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     2581 actgactcgc tgcgctcggt cgttcggctg cggcgagagg tatcagctca ctcaaaggcg
     2641 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2701 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2761 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2821 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2881 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2941 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3001 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3061 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3121 gcgaggta
//
