LOCUS       sequence-348478-        4683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PEAR-GFP3 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 4683)
  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 4683)
  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: pDAS12489_PEAR-GFP_2in1_2.0
FEATURES             Location/Qualifiers
     source          1..4683
                     /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        522..825
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        826..1029
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     980..1000
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1068..1077
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2177..2196)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2524..2545
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2588..2653
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2777..2898
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2814..2833)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2868..2887
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2957..3545)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3037..3056)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3716..4507,4508..4576))
                     /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     4339..4358
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4577..4681)
                     /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 cgagctggac ggcgacgtaa agttttagag ctagaaatag caagttaaaa taaggctagt
      361 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcggc tacgtgcagg taagtccgtt
      421 tacgtcgccg tccttttttc cgttacataa cttacggtaa atggcccaat tcgagtggct
      481 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccgttacata acttacggta
      541 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      601 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      661 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
      721 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
      781 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      841 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      901 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      961 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     1021 agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggtcgcc accatggtga
     1081 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgaactc gatggagatg
     1141 tgaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     1201 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     1261 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     1321 acttcttcaa gtccgccatg cccgaaggct acgtgcagac aagtccgttt acgtcgccgt
     1381 ccagctcatt ttttagttaa aatatgggaa agtttaaaag aaggaatgat gaataaacta
     1441 tacaaccact ttaaatatgt gttggtcccc tttaagtcag caagcagata tatcagatat
     1501 ggcagctaac ttaaggagtc cccgtggata ctggatgaat agcccccaat tccaccaatg
     1561 caagaaccaa cagtctattt ggtcttgatt aggacagtaa acttcctgca caccatagtc
     1621 tgtgcactaa acaggagtaa aacaccctta aggcatctct tacattggct gcattaaagt
     1681 gtgtcactta atgacatccg tgatatcagg gatagtttga cattggacat acagttctct
     1741 ctcagtgatt gtcacctgct tatatagtac gaggacccat atgtggtcaa tcactttctt
     1801 gttgccactc catgccaggg aagacatagt tacaatgagt tataattata aaagctcagc
     1861 cttgcatgag gatataaaat acatttgcca tacctctaca tatgccttag ttgcttttaa
     1921 gctttgtttg gaattgcaaa ggccaacttc ctaaacacag tttcaaagta ttcttttgtc
     1981 acacctttcc actggatact tgtattgtct gcaggaagag gccaccctgc ctatgcttct
     2041 ccttagggcc tcgttctgca gagtcagcat aaactcccag tctgctcaac caattctgtc
     2101 tgattagggc catatctcct taccatgagt tgatttttgt ttcttttttg agcaggagcg
     2161 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     2221 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     2281 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     2341 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     2401 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     2461 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     2521 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     2581 gtacaagtcc ggactcagat ctcgagctca agcttcgaat tctgcagtcg acggtaccgc
     2641 gggcccggga tccaccggat ctagataact gatcataatc agccatacca catttgtaga
     2701 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     2761 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     2821 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     2881 actcatcaat gtatcttagc tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2941 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3001 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3061 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3121 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3181 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3241 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3301 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3361 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3421 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3481 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3541 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3601 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3661 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3721 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3781 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3841 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3901 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3961 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4021 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4081 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4141 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4201 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4261 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4321 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4381 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4441 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4501 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4561 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4621 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4681 gca
//
