LOCUS       sequence-479114-        3131 bp    DNA     circular SYN 21-JUL-2025
DEFINITION  WT OrufIscB omegaRNA mammalian expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3131)
  AUTHORS   Kannan S, Altae-Tran H, Zhu S, Xu P, Strebinger D, Oshiro R, Faure 
            G, Moeller L, Pham J, Mears KS, Ni HM, Macrae RK, Zhang F
  TITLE     Evolution-guided protein design of IscB for persistent epigenome 
            editing in vivo.
  JOURNAL   Nat Biotechnol. 2025 May 7. doi: 10.1038/s41587-025-02655-3.
  PUBMED    40335752
REFERENCE   2  (bases 1 to 3131)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 21, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2025 May 7. doi: 10.1038/s41587-025-02655-3."
COMMENT     Sequence Label: pHS1376
FEATURES             Location/Qualifiers
     source          1..3131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     265..284
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     518..535
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    652..673
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(688..693,694..711,712..718)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    726..742
                     /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     731..753
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     750..766
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     750..766
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        780..1028
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     780..800
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     951..970
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(1281..1298)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1281..1297)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1290..1312)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1506..1525)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1625..1647
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1685..1703)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1771..1875
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1876..1944,1945..2736)
                     /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(2094..2113)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(2907..3131,1..364)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
       61 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
      121 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
      181 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
      241 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
      301 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
      361 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
      421 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
      481 agctgatacc tctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      541 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
      601 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
      661 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
      721 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccag
      781 agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga
      841 taattggaat taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa
      901 agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata
      961 tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac
     1021 gaaacaccat gtcttcctgg gacgaagaca agtcaataac ccatgactga agtcatgggc
     1081 ttgcagatgc aggtcctgat ggaagaaagg gttactgagc agagcagtga catgtcattc
     1141 gccgcggggt gattccaagc tccgcgctcc ggctagacat gcccatgcta tggaaacttt
     1201 aacggtatgt gcggttttcc gctcataccg gcttacaaca aataaggagt tattagtttt
     1261 ttggtaccga gctcgaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     1321 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     1381 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
     1441 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
     1501 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     1561 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     1621 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     1681 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     1741 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     1801 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     1861 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     1921 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     1981 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     2041 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     2101 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     2161 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     2221 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     2281 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     2341 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     2401 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     2461 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     2521 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     2581 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     2641 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     2701 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     2761 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     2821 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     2881 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     2941 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3001 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     3061 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     3121 ctgctaatcc t
//
