LOCUS       sequence-245794-        2311 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli  vector for CasX gRNA  --- Used for In Vitro Transcription.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2311)
  AUTHORS   Liu JJ, Orlova N, Oakes BL, Ma E, Spinner HB, Baney KLM, Chuck J, 
            Tan D, Knott GJ, Harrington LB, Al-Shayeb B, Wagner A, Brotzmann J, 
            Staahl BT, Taylor KL, Desmarais J, Nogales E, Doudna JA
  TITLE     CasX enzymes comprise a distinct family of RNA-guided genome 
            editors.
  JOURNAL   Nature. 2019 Feb 4. pii: 10.1038/s41586-019-0908-x. doi: 
            10.1038/s41586-019-0908-x.
  PUBMED    30718774
REFERENCE   2  (bases 1 to 2311)
  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. 
            2019 Feb 4. pii: 10.1038/s41586-019-0908-x. doi: 
            10.1038/s41586-019-0908-x."
COMMENT     Sequence Label: PBLO CasX-gRNA
FEATURES             Location/Qualifiers
     source          1..2311
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..35
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     primer_bind     complement(266..283)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(437..1025)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(517..536)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1196..1987,1988..2056))
                     /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     1819..1838
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2057..2161)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2229..2247
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2251..2294)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 ttgacagcta gctcagtcct aggtataata ctagtacatc tggcgcgttt attccattac
       61 tttggagcca gtcccagcga ctatgtcgta tggacgaagc gcttatttat cggagagaaa
      121 ccgataagta aaacgcatca aagctggagt tgtcccaatt cttctagagt cacactggct
      181 caccttcggg tgggccctag agtcacactg gctcaccttc gggtgtttct gcgggatcct
      241 tactcgagtc tagactgcag gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
      301 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
      361 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
      421 ccgcgttgct ggcgtttttc cacaggctcc gcccccctga cgagcatcac aaaaatcgac
      481 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
      541 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
      601 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
      661 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
      721 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
      781 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
      841 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
      901 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
      961 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     1021 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     1081 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     1141 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     1201 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     1261 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     1321 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     1381 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     1441 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     1501 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     1561 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     1621 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     1681 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     1741 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     1801 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     1861 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     1921 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     1981 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     2041 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     2101 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     2161 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     2221 cctataaaaa taggcgtatc acgaggcaga atttcagata aaaaaaatcc ttagctttcg
     2281 ctaaggatga tttctggaat tctaaagatc t
//
