LOCUS       sequence-150232-        2436 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning vector for expression of LbCpf1 crRNA. It contains BsmB1 
            site for cloning..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2436)
  AUTHORS   Kim D, Kim J, Hur JK, Been KW, Yoon SH, Kim JS
  TITLE     Genome-wide analysis reveals specificities of Cpf1 endonucleases in 
            human cells.
  JOURNAL   Nat Biotechnol. 2016 Jun 6. doi: 10.1038/nbt.3609.
  PUBMED    27272384
REFERENCE   2  (bases 1 to 2436)
  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. 2016 Jun 6. doi: 10.1038/nbt.3609."
COMMENT     Sequence Label: pU6-Lb-crRNA
FEATURES             Location/Qualifiers
     source          1..2436
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        84..332
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     84..104
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     255..274
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(510..527)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(681..1269)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(761..780)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1440..2231,2232..2300))
                     /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     2063..2082
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2301..2405)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgaagact caattgtcga ttagtgaacg gatctcgacg gtatcgatca cgagactagc
       61 ctcgagcggc cgcccccttc accgagggcc tatttcccat gattccttca tatttgcata
      121 tacgatacaa ggctgttaga gagataattg gaattaattt gactgtaaac acaaagatat
      181 tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat
      241 tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg
      301 ctttatatat cttgtggaaa ggacgaaaca ccgaatttct actaagtgta gattgagacg
      361 ggatccatcc gtctcctttt ttctagattc gcgatgtacg ggccagatat acgcgttgac
      421 attgattatt gactagttgt cttcctgcat taatgaatcg gccaacgcgc ggggagaggc
      481 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
      541 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
      601 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      661 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
      721 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
      781 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
      841 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
      901 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
      961 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     1021 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     1081 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     1141 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     1201 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     1261 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     1321 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     1381 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     1441 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     1501 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     1561 agtgctgcaa tgataccgcg agatccacgc tcaccggctc cagatttatc agcaataaac
     1621 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     1681 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     1741 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     1801 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     1861 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     1921 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     1981 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     2041 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     2101 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     2161 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     2221 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     2281 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     2341 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     2401 ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
