LOCUS       sequence-449445-        3128 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid codes for the guide of the OgeuIscB with a AGC 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 AGC RNA
FEATURES             Location/Qualifiers
     source          1..3128
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     158..177
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     411..428
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    545..566
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(581..586,587..604,605..611)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    619..635
                     /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     624..646
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     643..659
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     643..659
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        673..921
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     673..693
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     844..863
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(1171..1188)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1171..1187)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1180..1202)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1396..1415)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1515..1537
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1575..1593)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1661..1765
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1766..1834,1835..2626)
                     /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(1984..2003)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(2797..3128,1..257)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
       61 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
      121 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
      181 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
      241 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
      301 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
      361 ccgcctttga gtgagctgat acctctcgcc gcagccgaac gaccgagcgc agcgagtcag
      421 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
      481 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gaacgcaacg
      541 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
      601 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
      661 atgattacgc cagagggcct atttcccatg attccttcat atttgcatat acgatacaag
      721 gctgttagag agataattgg aattaatttg actgtaaaca caaagatatt agtacaaaat
      781 acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa
      841 atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc
      901 ttgtggaaag gacgaaacac cgagcagcag cagcagcagc ggctcttcca actttatggt
      961 tgcgaccgta ggttgaaaga gcacaggctg agacattcgt aaggccgaaa gaccggacgc
     1021 accctgggat ttccccagtc cccggaactg catagcggat gccagttgat ggagcaatct
     1081 atcagataag ccagggggaa caatcacctc tctgtatcag agagagtttt acaaaaggag
     1141 gaacggtttt ttggtaccga gctcgaattc actggccgtc gttttacaac gtcgtgactg
     1201 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     1261 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     1321 cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     1381 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     1441 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     1501 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     1561 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     1621 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     1681 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     1741 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     1801 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     1861 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     1921 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     1981 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     2041 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     2101 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     2161 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     2221 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     2281 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     2341 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     2401 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     2461 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     2521 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     2581 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     2641 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     2701 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2761 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2821 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2881 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2941 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3001 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3061 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3121 gggttcgt
//
