LOCUS       Exported                2908 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Backbone for cloning LwCas13a guides under U6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pC016 - LwCas13a guide expression backbone with U6 promoter
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2908)
  AUTHORS   Abudayyeh OO, Gootenberg JS, Essletzbichler P, Han S, Joung J, 
            Belanto JJ, Verdine V, Cox DBT, Kellner MJ, Regev A, Lander ES, 
            Voytas DF, Ting AY, Zhang F
  TITLE     RNA targeting with CRISPR-Cas13.
  JOURNAL   Nature. 2017 Oct 4. doi: 10.1038/nature24049.
  PUBMED    28976959
REFERENCE   2  (bases 1 to 2908)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2017 Oct 4. doi: 10.1038/nature24049."
FEATURES             Location/Qualifiers
     source          1..2908
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      138..726
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     627..646
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        789..1029
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     789..809
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     960..979
                     /label=LKO.1 5
                     /note="Human U6 promoter, forward primer"
     rep_origin      1173..1628
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1260..1279)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1470..1491
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1645..1664)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1764..1786
                     /label=pGEX 3
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1824..1842)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1910..2014
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2015..2875
                     /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(2233..2252)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"