LOCUS       Exported                2301 bp DNA     circular SYN 17-DEC-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2301)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 17, 2021 from SnapGene Viewer 6.0.0
            https://www.snapgene.com
COMMENT     LOCUS       Exported                3004 bp ds-DNA     circular SYN 
            18-OCT-2019

            DEFINITION  Prime editing in mammalian cells.

            ACCESSION   .

            VERSION     .

            KEYWORDS    pU6-pegRNA-GG-acceptor

            SOURCE      synthetic DNA construct

              ORGANISM  synthetic DNA construct

            REFERENCE   1  (bases 1 to 3004)

              TITLE     Liu Lab plasmids

              JOURNAL   Unpublished

            REFERENCE   2  (bases 1 to 3004)

              AUTHORS   .

              TITLE     Direct Submission

              JOURNAL   Exported Oct 18, 2019 from SnapGene Server 1.1.58

                        http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..2301
                     /label=synthetic DNA construct
                     /note="color: #000000"
                     /organism="synthetic DNA construct"
     promoter        1..241
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
                     /note="color: #8080ff; direction: RIGHT"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
                     /note="color: #ff8080; direction: RIGHT"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
                     /note="color: #ff8080; direction: RIGHT"
     misc_feature    250..270
                     /label=pegRNA-Spacer
                     /note="pegRNA-Spacer"
                     /note="color: #804000"
     misc_feature    271..346
                     /label=pegRNA-scaffold
                     /note="pegRNA-scaffold"
                     /note="color: #804000"
     misc_RNA        271..346
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
                     /note="color: #00ccff; direction: RIGHT"
     misc_feature    347..367
                     /label=pegRNA-Extension-PBS9-RTT12
                     /note="pegRNA-Extension-PBS9-RTT12"
                     /note="color: #804000"
     rep_origin      complement(471..1059)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #c0c0c0"
     primer_bind     complement(551..570)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="color: #ff8080; direction: LEFT"
     CDS             complement(1230..2090)
                     /codon_start=1
                     /label=bla
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="color: #e8fafd"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1853..1872
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
                     /note="color: #ff8080; direction: RIGHT"
     promoter        complement(2091..2195)
                     /label=bla
                     /note="color: #8080ff; direction: LEFT"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg attgcaaaga ttgctgacta gttttagagc tagaaatagc aagttaaaat
      301 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgcgcaa tgctgtagtc
      361 agcaatcttt ttttaagctt gggccgctcg aggtacctct ctacatatga catgtgagca
      421 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
      481 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
      541 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
      601 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
      661 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
      721 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
      781 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
      841 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
      901 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
      961 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     1021 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     1081 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     1141 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     1201 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     1261 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     1321 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agatccacgc
     1381 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     1441 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     1501 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     1561 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     1621 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     1681 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     1741 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     1801 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     1861 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     1921 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     1981 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     2041 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     2101 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     2161 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     2221 gacgtcgcta gctgtacaaa aaagcaggct ttaaaggaac caattcagtc gactggatcc
     2281 ggtaccaagg tcgggcagga a
//
