LOCUS       sequence-345280-        2962 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  hU6-BpiI-Cas13bt3-DR (crRNA expression).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2962)
  AUTHORS   Kannan S, Altae-Tran H, Jin X, Madigan VJ, Oshiro R, Makarova KS, 
            Koonin EV, Zhang F
  TITLE     Compact RNA editors with small Cas13 proteins.
  JOURNAL   Nat Biotechnol. 2021 Aug 30. pii: 10.1038/s41587-021-01030-2. doi: 
            10.1038/s41587-021-01030-2.
  PUBMED    34462587
REFERENCE   2  (bases 1 to 2962)
  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 
            Biotechnol. 2021 Aug 30. pii: 10.1038/s41587-021-01030-2. doi: 
            10.1038/s41587-021-01030-2."
COMMENT     Sequence Label: pAB1620 hU6-BpiI-Cas13bt3-DR
FEATURES             Location/Qualifiers
     source          1..2962
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    102..123
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(138..143,144..161,162..168)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    176..192
                     /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     181..203
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     200..216
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     200..216
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        230..478
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     230..250
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     401..420
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(562..579)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(562..578)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(571..593)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(787..806)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     906..928
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(966..984)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1052..1156
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1157..1225,1226..2017)
                     /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(1375..1394)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2188..2776
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2677..2696
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2930..2947
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
       61 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
      121 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
      181 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccag agggcctatt
      241 tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat
      301 taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt
      361 tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt
      421 aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccat
      481 gtcttcctgg gacgaagaca agctggagca gcccccgatt tgtggggtga ttacagcttt
      541 tttggtaccg agctcgaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
      601 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
      661 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg
      721 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac
      781 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
      841 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
      901 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
      961 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     1021 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     1081 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     1141 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     1201 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     1261 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     1321 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     1381 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     1441 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     1501 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     1561 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     1621 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     1681 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     1741 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     1801 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     1861 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     1921 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     1981 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     2041 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     2101 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2161 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2221 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2281 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     2341 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     2401 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     2461 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     2521 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     2581 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     2641 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     2701 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     2761 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     2821 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     2881 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     2941 gagcgaggaa gcggaagagc gc
//
