LOCUS       sequence-341937-        3132 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  human U6-driven expression of OgeuIscB omegaRNA scaffold with BpiI 
            site for guide cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3132)
  AUTHORS   Altae-Tran H, Kannan S, Demircioglu FE, Oshiro R, Nety SP, McKay LJ,
            Dlakic M, Inskeep WP, Makarova KS, Macrae RK, Koonin EV, Zhang F
  TITLE     The widespread IS200/IS605 transposon family encodes diverse 
            programmable RNA-guided endonucleases.
  JOURNAL   Science. 2021 Oct;374(6563):57-65. doi: 10.1126/science.abj6856. 
            Epub 2021 Sep 9.
  PUBMED    34591643
REFERENCE   2  (bases 1 to 3132)
  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"; doi: 
            "10.1126/science.abj6856"; journalName: "Science"; date: "2021-10"; 
            volume: "374"; issue: "6563"; pages: "57-65"
COMMENT     Sequence Label: pHS0812 hU6-BpiI-OgeuIscB omegaRNA scaffold
FEATURES             Location/Qualifiers
     source          1..3132
                     /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(1282..1299)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1282..1298)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1291..1313)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1507..1526)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1626..1648
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1686..1704)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1772..1876
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1877..1945,1946..2737)
                     /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(2095..2114)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(2908..3132,1..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 gctcgccgca 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 aggctcttcc aactttatgg ttgcgaccgt
     1081 aggttgaaag agcacaggct gagacattcg taaggccgaa agaccggacg caccctggga
     1141 tttccccagt ccccggaact gcatagcgga tgccagttga tggagcaatc tatcagataa
     1201 gccaggggga acaatcacct ctctgtatca gagagagttt tacaaaagga ggaacggttt
     1261 tttggtaccg agctcgaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     1321 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     1381 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg
     1441 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac
     1501 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     1561 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     1621 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     1681 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     1741 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     1801 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     1861 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     1921 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     1981 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     2041 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     2101 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     2161 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     2221 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     2281 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     2341 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     2401 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     2461 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     2521 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     2581 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     2641 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     2701 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     2761 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     2821 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2881 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2941 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3001 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3061 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3121 tctgctaatc ct
//
