LOCUS       sequence-348471-        4681 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PEAR-GFP plasmid with a pegRNA that targets its own plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4681)
  AUTHORS   Simon DA, Talas A, Kulcsar PI, Biczok Z, Krausz SL, Varady G, Welker
            E
  TITLE     PEAR, a flexible fluorescent reporter for the identification and 
            enrichment of successfully prime edited cells.
  JOURNAL   Elife. 2022 Feb 23;11. pii: 69504. doi: 10.7554/eLife.69504.
  PUBMED    35196219
REFERENCE   2  (bases 1 to 4681)
  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: "Elife."; 
            date: "2022-02-23"; volume: "11. pii: 69504. doi"; pages: " 
            10.7554/eLife.69504"
COMMENT     Sequence Label: pDAS12342_PEAR-GFP_2in1
FEATURES             Location/Qualifiers
     source          1..4681
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        53..301
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     53..73
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     224..243
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        322..397
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        520..823
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        824..1027
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     978..998
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1066..1075
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1117..1138)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2175..2194)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2522..2543
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2586..2651
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2775..2896
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2812..2831)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2866..2885
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2955..3543)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3035..3054)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3714..4505,4506..4574))
                     /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     4337..4356
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4575..4679)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 catttccccg aaaagtgcca cctgacgtcc agcagagatc cactttggcg ccgagggcct
       61 atttcccatg attccttcat atttgcatat acgatacaag gctgttagag agataattgg
      121 aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta gaaagtaata
      181 atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc atatgcttac
      241 cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag gacgaaacac
      301 cgaatgtcaa acccgcagtc tgttttagag ctagaaatag caagttaaaa taaggctagt
      361 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcggc tacgtgcagg taagtcctag
      421 actgcgggtt tttttttccg ttacataact tacggtaaat ggcccaattc gagtggctcc
      481 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc gttacataac ttacggtaaa
      541 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      601 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      661 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      721 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      781 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      841 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      901 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      961 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     1021 cagagctggt ttagtgaacc gtcagatccg ctagcgctac cggtcgccac catggtgagc
     1081 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     1141 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     1201 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     1261 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     1321 ttcttcaagt ccgccatgcc cgaaggctac gtgcagacaa gtcctagact gcgggtttga
     1381 cattcatttt ttagttaaaa tatgggaaag tttaaaagaa ggaatgatga ataaactata
     1441 caaccacttt aaatatgtgt tggtcccctt taagtcagca agcagatata tcagatatgg
     1501 cagctaactt aaggagtccc cgtggatact ggatgaatag cccccaattc caccaatgca
     1561 agaaccaaca gtctatttgg tcttgattag gacagtaaac ttcctgcaca ccatagtctg
     1621 tgcactaaac aggagtaaaa cacccttaag gcatctctta cattggctgc attaaagtgt
     1681 gtcacttaat gacatccgtg atatcaggga tagtttgaca ttggacatac agttctctct
     1741 cagtgattgt cacctgctta tatagtacga ggacccatat gtggtcaatc actttcttgt
     1801 tgccactcca tgccagggaa gacatagtta caatgagtta taattataaa agctcagcct
     1861 tgcatgagga tataaaatac atttgccata cctctacata tgccttagtt gcttttaagc
     1921 tttgtttgga attgcaaagg ccaacttcct aaacacagtt tcaaagtatt cttttgtcac
     1981 acctttccac tggatacttg tattgtctgc aggaagaggc caccctgcct atgcttctcc
     2041 ttagggcctc gttctgcaga gtcagcataa actcccagtc tgctcaacca attctgtctg
     2101 attagggcca tatctcctta ccatgagttg atttttgttt cttttttgag caggagcgca
     2161 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     2221 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     2281 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     2341 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     2401 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     2461 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     2521 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     2581 acaagtccgg actcagatct cgagctcaag cttcgaattc tgcagtcgac ggtaccgcgg
     2641 gcccgggatc caccggatct agataactga tcataatcag ccataccaca tttgtagagg
     2701 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     2761 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     2821 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     2881 tcatcaatgt atcttagctg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2941 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3001 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3061 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3121 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3181 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3241 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3301 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3361 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3421 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3481 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3541 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3601 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     3661 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     3721 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     3781 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     3841 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3901 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3961 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4021 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4081 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4141 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4201 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4261 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4321 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4381 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4441 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     4501 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     4561 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     4621 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     4681 a
//
