LOCUS       sequence-348473-        5528 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PEAR-GFP target plasmid with split EGFP and disrupted 5' splice 
            site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5528)
  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 5528)
  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: pDAS12125_PEAR-GFP
FEATURES             Location/Qualifiers
     source          1..5528
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1716..1735)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2063..2084
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2127..2192
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2316..2437
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2353..2372)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2407..2426
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2444..2899)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2581..2602)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2793..2812
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2926..3030
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3032..3389
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3032..3223
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3055..3075)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3240..3375
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3424..4218
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3424..4218
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3478..3497)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4088..4107
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4406..4425)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4450..4497
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4826..5414
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5315..5334
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtgcagaca
      901 agtcctagac tgcgggtttg acattcattt tttagttaaa atatgggaaa gtttaaaaga
      961 aggaatgatg aataaactat acaaccactt taaatatgtg ttggtcccct ttaagtcagc
     1021 aagcagatat atcagatatg gcagctaact taaggagtcc ccgtggatac tggatgaata
     1081 gcccccaatt ccaccaatgc aagaaccaac agtctatttg gtcttgatta ggacagtaaa
     1141 cttcctgcac accatagtct gtgcactaaa caggagtaaa acacccttaa ggcatctctt
     1201 acattggctg cattaaagtg tgtcacttaa tgacatccgt gatatcaggg atagtttgac
     1261 attggacata cagttctctc tcagtgattg tcacctgctt atatagtacg aggacccata
     1321 tgtggtcaat cactttcttg ttgccactcc atgccaggga agacatagtt acaatgagtt
     1381 ataattataa aagctcagcc ttgcatgagg atataaaata catttgccat acctctacat
     1441 atgccttagt tgcttttaag ctttgtttgg aattgcaaag gccaacttcc taaacacagt
     1501 ttcaaagtat tcttttgtca cacctttcca ctggatactt gtattgtctg caggaagagg
     1561 ccaccctgcc tatgcttctc cttagggcct cgttctgcag agtcagcata aactcccagt
     1621 ctgctcaacc aattctgtct gattagggcc atatctcctt accatgagtt gatttttgtt
     1681 tcttttttga gcaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     1741 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     1801 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     1861 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     1921 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     1981 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     2041 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     2101 ctctcggcat ggacgagctg tacaagtccg gactcagatc tcgagctcaa gcttcgaatt
     2161 ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc tagataactg atcataatca
     2221 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     2281 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     2341 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     2401 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat
     2461 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     2521 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     2581 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     2641 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     2701 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     2761 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     2821 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     2881 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     2941 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3001 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     3061 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3121 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3181 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3241 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3301 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3361 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     3421 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3481 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3541 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3601 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3661 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3721 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3781 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3841 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3901 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     3961 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4021 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4081 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4141 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4201 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4261 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4321 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4381 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     4441 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     4501 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     4561 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     4621 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     4681 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4741 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4801 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4861 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4921 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4981 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5041 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5101 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5161 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5221 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5281 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5341 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5401 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5461 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     5521 ccatgcat
//
