LOCUS       sequence-449443-        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     complement(37..56)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      850..1438
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1339..1358
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1592..1609
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1726..1747
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1762..1767,1768..1785,1786..1792)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1800..1816
                     /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     1805..1827
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1824..1840
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1824..1840
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1854..2102
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1854..1874
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2025..2044
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(2352..2369)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2352..2368)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2361..2383)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2577..2596)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2696..2718
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2756..2774)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2842..2946
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2947..3015,3016..3128,1..679)
                     /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"
ORIGIN
        1 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
       61 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
      121 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
      181 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
      241 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
      301 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
      361 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
      421 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
      481 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
      541 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
      601 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
      661 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
      721 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
      781 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
      841 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
      901 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
      961 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     1021 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1081 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1141 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1201 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     1261 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     1321 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     1381 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     1441 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     1501 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     1561 tacctctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     1621 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     1681 cgacaggttt cccgactgga aagcgggcag tgaacgcaac gcaattaatg tgagttagct
     1741 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     1801 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccagagggcc
     1861 tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga gagataattg
     1921 gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt agaaagtaat
     1981 aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta
     2041 ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttgtggaaa ggacgaaaca
     2101 ccgcagcagc agcagcagca gggctcttcc aactttatgg ttgcgaccgt aggttgaaag
     2161 agcacaggct gagacattcg taaggccgaa agaccggacg caccctggga tttccccagt
     2221 ccccggaact gcatagcgga tgccagttga tggagcaatc tatcagataa gccaggggga
     2281 acaatcacct ctctgtatca gagagagttt tacaaaagga ggaacggttt tttggtaccg
     2341 agctcgaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     2401 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     2461 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg
     2521 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     2581 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     2641 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     2701 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     2761 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     2821 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     2881 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     2941 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     3001 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     3061 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     3121 cgccccga
//
