LOCUS       sequence-150238-        2435 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning vector for expression of FnCpf1 crRNA. It contains BsmB1 
            site for cloning..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2435)
  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 2435)
  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-Fn-crRNA
FEATURES             Location/Qualifiers
     source          1..2435
                     /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(509..526)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(680..1268)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(760..779)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1439..2230,2231..2299))
                     /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     2062..2081
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2300..2404)
                     /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 actgttgtag attgagacgg
      361 gatccatccg tctccttttt tctagattcg cgatgtacgg gccagatata cgcgttgaca
      421 ttgattattg actagttgtc ttcctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
      481 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
      541 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
      601 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
      661 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
      721 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
      781 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
      841 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
      901 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
      961 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     1021 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     1081 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     1141 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     1201 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     1261 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     1321 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     1381 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     1441 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     1501 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     1561 gtgctgcaat gataccgcga gatccacgct caccggctcc agatttatca gcaataaacc
     1621 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     1681 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     1741 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     1801 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     1861 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     1921 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     1981 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     2041 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     2101 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     2161 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     2221 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     2281 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     2341 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     2401 cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
