LOCUS       Exported                2307 bp DNA     circular SYN 17-DEC-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2307)
  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              JOURNAL   Unpublished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..2307
                     /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"
     promoter        1..241
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
                     /note="color: #ffffff; 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_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..373
                     /label=pegRNA-Extension-PBS15-RTT12
                     /note="pegRNA-Extension-PBS15-RTT12"
                     /note="color: #804000"
     rep_origin      complement(477..1065)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #c0c0c0"
     rep_origin      complement(477..1065)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     complement(557..576)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="color: #ff8080; direction: LEFT"
     CDS             complement(1236..2096)
                     /codon_start=1
                     /label=bla
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="color: #e8fafd"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             complement(1236..2096)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 1236 .. 2027 / #ccffcc
                        2: 2028 .. 2096 / #ccffcc / signal sequence
                     Cleavage site after base 2027"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1859..1878
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
                     /note="color: #ff8080; direction: RIGHT"
     promoter        complement(2097..2201)
                     /label=bla
                     /note="color: #8080ff; direction: LEFT"
     promoter        complement(2097..2201)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; 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 gcattttttt aagcttgggc cgctcgaggt acctctctac atatgacatg
      421 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      481 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      541 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      601 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      661 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      721 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      781 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      841 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
      901 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
      961 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     1021 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     1081 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     1141 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     1201 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     1261 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     1321 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagat
     1381 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     1441 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     1501 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     1561 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     1621 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     1681 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     1741 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     1801 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     1861 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     1921 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     1981 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     2041 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     2101 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     2161 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     2221 ccacctgacg tcgctagctg tacaaaaaag caggctttaa aggaaccaat tcagtcgact
     2281 ggatccggta ccaaggtcgg gcaggaa
//
