LOCUS       sequence-443130-        2747 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  human U6-driven expression of omega-RNA-v2 scaffold with BsmBI for 
            guide cloing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2747)
  AUTHORS   Yan H, Tan X, Zou S, Sun Y, Ke A, Tang W
  TITLE     Assessing and engineering the IscB-omegaRNA system for programmed 
            genome editing.
  JOURNAL   Nat Chem Biol. 2024 Jul 8. doi: 10.1038/s41589-024-01669-3.
  PUBMED    38977787
REFERENCE   2  (bases 1 to 2747)
  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: "Nat Chem 
            Biol. 2024 Jul 8. doi: 10.1038/s41589-024-01669-3."
COMMENT     Sequence Label: OgeuIscB omega-RNA-v2
FEATURES             Location/Qualifiers
     source          1..2747
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(117..221)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(296..315)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     320..342
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     334..351
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     335..351
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(580..828)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(638..657)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(808..828)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(842..858)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(842..858)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(855..877)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    866..882
                     /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(890..896,897..914,915..920))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    935..956
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1073..1090)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1244..1832)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1324..1343)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2003..2747,1..47,48..116))
                     /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     2626..2645
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
       61 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
      121 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
      181 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
      241 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
      301 tctgctctga tgccgagctc ccagtcacga cgttgtaaaa cgacggccag tgaattcgag
      361 ctcggtacca aaaaaaactc tctctgtttc cagagaggtg attgttcccc ctggcttatc
      421 tgattttcat caactgacat ccgctatgca gttccgggga ctggggaaat cccagggtgc
      481 gtccggcctt tcggccttac gaatgtctca gcctgtgctc tttcaacctt ttcaagttga
      541 aagagcctga gacgatctcg ttcactcttt tcgtctctcg gtgtttcgtc ctttccacaa
      601 gatatataaa gccaagaaat cgaaatactt tcaagttacg gtaagcatat gatagtccat
      661 tttaaaacat aattttaaaa ctgcaaacta cccaagaaat tattactttc tacgtcacgt
      721 attttgtact aatatctttg tgtttacagt caaattaatt ccaattatct ctctaacagc
      781 cttgtatcgt atatgcaaat atgaaggaat catgggaaat aggccctctg gcgtaatcat
      841 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
      901 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
      961 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     1021 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     1081 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     1141 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     1201 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     1261 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     1321 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     1381 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     1441 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     1501 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     1561 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     1621 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     1681 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     1741 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     1801 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     1861 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     1921 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     1981 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     2041 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     2101 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     2161 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     2221 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     2281 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     2341 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     2401 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     2461 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     2521 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     2581 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     2641 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     2701 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgca
//
