LOCUS       sequence-176607-        2766 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Backbone for cloning LwCas13a guides under tRNAval promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2766)
  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 2766)
  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: "Nature. 
            2017 Oct 4. doi: 10.1038/nature24049."
COMMENT     Sequence Label: pC017 - LwCas13a guide expression backbone wit...
FEATURES             Location/Qualifiers
     source          1..2766
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      159..747
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     648..667
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      1052..1507
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1139..1158)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1349..1370
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1524..1543)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1643..1665
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1703..1721)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1789..1893
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1894..1962,1963..2754)
                     /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(2112..2131)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
       61 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
      121 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
      181 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
      241 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
      301 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
      361 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
      421 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
      481 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
      541 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
      601 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
      661 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
      721 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
      781 gccttttgct ggccttttgc tcagctagcc aggactagtc ttttaggtca aaaagaagaa
      841 gctttgtaac cgttggtttc cgtagtgtag tggttatcac gttcgcctaa cacgcgaaag
      901 gtccccggtt cgaaggattt agactacccc aaaaacgaag gggactaaaa cgggtcttcg
      961 agaagacctt tttttttgaa ttctgatgcg gtattttctc cttacgcatc tgtgcggtat
     1021 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     1081 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct
     1141 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     1201 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     1261 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     1321 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     1381 aactctatct cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg
     1441 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     1501 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     1561 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     1621 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     1681 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     1741 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     1801 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     1861 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     1921 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     1981 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     2041 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     2101 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     2161 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     2221 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     2281 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     2341 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     2401 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     2461 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     2521 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     2581 gctgataaat ctggagccgg tgagcgtgga agccgcggta tcattgcagc actggggcca
     2641 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     2701 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     2761 gaccaa
//
