LOCUS       sequence-443113-        2747 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  human U6-driven expression of OgeuIscB omegaRNA-v13 (omega-RNA-v13) 
            scaffold with BsmBI for guide cloning.
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-v13
FEATURES             Location/Qualifiers
     source          1..2747
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(125..229)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(304..323)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     328..350
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     342..359
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     343..359
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(588..836)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(646..665)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(816..836)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(850..866)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(850..866)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(863..885)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    874..890
                     /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(898..904,905..922,923..928))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    943..964
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1081..1098)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1252..1840)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1332..1351)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2011..2747,1..55,56..124))
                     /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     2634..2653
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
       61 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
      121 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
      181 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
      241 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
      301 cagtacaatc tgctctgatg ccgagctccc agtcacgacg ttgtaaaacg acggccagtg
      361 aattcgagct cggtaccaaa aaaaactctc tctgtttcca gagaggtgat tgttccccct
      421 ggcttatctg attttcatca actgacatcc gctatgcagt tccggggact ggggaaatcc
      481 cagggtgcgt ccggcctttc ggccttacga atgtctcagc ctgtgctctt tcaacctttt
      541 caagttggaa gagcctgaga cgatctcgtt cactcttttc gtctctcggt gtttcgtcct
      601 ttccacaaga tatataaagc caagaaatcg aaatactttc aagttacggt aagcatatga
      661 tagtccattt taaaacataa ttttaaaact gcaaactacc caagaaatta ttactttcta
      721 cgtcacgtat tttgtactaa tatctttgtg tttacagtca aattaattcc aattatctct
      781 ctaacagcct tgtatcgtat atgcaaatat gaaggaatca tgggaaatag gccctctggc
      841 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
      901 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
      961 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     1021 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     1081 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1141 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1201 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1261 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1321 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1381 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1441 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1501 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1561 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1621 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1681 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1741 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1801 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     1861 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     1921 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     1981 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2041 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2101 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     2161 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2221 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2281 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     2341 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     2401 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     2461 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     2521 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     2581 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     2641 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     2701 actctcaagg atcttaccgc tgttgagatc cagttcgatg taaccca
//
