LOCUS       sequence-420055-        5391 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Express PUFc_NES-ADARdd(E488Q).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5391)
  AUTHORS   Liu Z, Jillette N, Robson P, Cheng AW
  TITLE     Simultaneous multifunctional transcriptome engineering by CRISPR RNA
            scaffold.
  JOURNAL   Nucleic Acids Res. 2023 Aug 11;51(14):e77. doi: 10.1093/nar/gkad547.
  PUBMED    37395412
REFERENCE   2  (bases 1 to 5391)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "2023-08-11"; volume: "51(14):e77. doi"; pages: "
            10.1093/nar/gkad547"
COMMENT     Sequence Label: pmax-PUFc_NES-ADARdd(E488Q)
FEATURES             Location/Qualifiers
     source          1..5391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          813..945
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     protein_bind    960..984
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2193..2219
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     protein_bind    complement(3420..3444)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3497..3538
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3564..3583)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    3593..3674
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     3618..3637
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3872..4460)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3952..3971)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4541..5335)
                     /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             complement(4541..5335)
                     /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(4652..4671)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5262..5281
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagg
      721 tcgtttagtg aaccgtcaga tcactagtag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctc gactgatcac aggtaagtat caaggttaca agacaggttt
      841 aaggaggcca atagaaactg ggcttgtcga gacagagaag attcttgcgt ttctgatagg
      901 cacctattgg tcttactgac atccactttg cctttctctc cacagggaaa aaacaattga
      961 caagtttgta caaaaaagca ggctccgaat tcaccggtgc cgccaccatg aacgttggtg
     1021 gtggtggaag cgggggtggg ggcagcggtg gagggggaag cgggcgcgcc tctagaggcc
     1081 gcagccgcct tttggaagat tttcgaaaca accggtaccc caatttacaa ctgcgggaga
     1141 ttgctggaca tataatggaa ttttcccaag accagcatgg gtccagattc attcagctga
     1201 aactggagcg tgccacacca gctgagcgcc agcttgtctt caatgaaatc ctccaggctg
     1261 cctaccaact catggtggat gtgtttggta attacgtcat tcagaagttc tttgaatttg
     1321 gcagtcttga acagaagctg gctttggcag aacggattcg aggccacgtc ctgtcattgg
     1381 cactacagat gtatggcagc cgtgttatcg agaaagctct tgagtttatt ccttcagacc
     1441 agcagaatga gatggttcgg gaactagatg gccatgtctt gaagtgtgtg aaagatcaga
     1501 atggcaatca cgtggttcag aaatgcattg aatgtgtaca gccccagtct ttgcaattta
     1561 tcatcgacgc gtttaaggga caggtatttg ccttatccac acatccttat ggctgccgag
     1621 tgattcagag aatcctggag cactgtctcc ctgaccagac actccctatt ttagaggagc
     1681 ttcaccagca cacagagcag ctggtacagg atcaatatgg aagttatgta atcgaacatg
     1741 tactggagca cggtcgtcct gaggataaaa gcaaaattgt agcagaaatc cgaggcaatg
     1801 tacttgtatt gagtcagcac aaatttgcaa acaatgttgt gcagaagtgt gttactcacg
     1861 cctcacgtac ggagcgcgct gtgctcatcg acgaggtgtg caccatgaac gacggtcccc
     1921 acagtgcctt atacaccatg atgaaggacc agtatgccaa ctacgtggtc cagaagatga
     1981 ttgacgtggc ggagccaggc cagcggaaga tcgtcatgca taagatccgg ccccacatcg
     2041 caactcttcg taagtacacc tatggcaagc acattctggc caagctggag aagtactaca
     2101 tgaagaacgg tgttgactta gggggcggcc gcgggggagg cggttccggt ggcggcggaa
     2161 gcggaggtgg aggatcaggg ccggccctgc agctgcctcc acttgaaaga ctgacactgg
     2221 gatccggagg aggtggaagc cagctgcatt taccgcaggt tttagctgac gctgtctcac
     2281 gcctggtcct gggtaagttt ggtgacctga ccgacaactt ctcctcccct cacgctcgca
     2341 gaaaagtgct ggctggagtc gtcatgacaa caggcacaga tgttaaagat gccaaggtga
     2401 taagtgtttc tacaggaaca aaatgtatta atggtgaata catgagtgat cgtggccttg
     2461 cattaaatga ctgccatgca gaaataatat ctcggagatc cttgctcaga tttctttata
     2521 cacaacttga gctttactta aataacaaag atgatcaaaa aagatccatc tttcagaaat
     2581 cagagcgagg ggggtttagg ctgaaggaga atgtccagtt tcatctgtac atcagcacct
     2641 ctccctgtgg agatgccaga atcttctcac cacatgagcc aatcctggaa gaaccagcag
     2701 atagacaccc aaatcgtaaa gcaagaggac agctacggac caaaatagag tctggtcagg
     2761 ggacgattcc agtgcgctcc aatgcgagca tccaaacgtg ggacggggtg ctgcaagggg
     2821 agcggctgct caccatgtcc tgcagtgaca agattgcacg ctggaacgtg gtgggcatcc
     2881 agggatcact gctcagcatt ttcgtggagc ccatttactt ctcgagcatc atcctgggca
     2941 gcctttacca cggggaccac ctttccaggg ccatgtacca gcggatctcc aacatagagg
     3001 acctgccacc tctctacacc ctcaacaagc ctttgctcag tggcatcagc aatgcagaag
     3061 cacggcagcc agggaaggcc cccaacttca gtgtcaactg gacggtaggc gactccgcta
     3121 ttgaggtcat caacgccacg actgggaagg atgagctggg ccgcgcgtcc cgcctgtgta
     3181 agcacgcgtt gtactgtcgc tggatgcgtg tgcacggcaa ggttccctcc cacttactac
     3241 gctccaagat taccaagccc aacgtgtacc atgagtccaa gctggcggca aaggagtacc
     3301 aggccgccaa ggcgcgtctg ttcacagcct tcatcaaggc ggggctgggg gcctgggtgg
     3361 agaagcccac cgagcaggac cagttctcac tcacgaacgt ttgattaatt aagaattcga
     3421 cccagctttc ttgtacaaag tggttgatat ccagcacagt ggcggccgct cgagtctaga
     3481 gggcccgcgg ttcgaaggta agcctatccc taaccctctc ctcggtctcg attctacgcg
     3541 taccggttag caattgtttt ttcgatgagt ttggacaaac cacaactaga atgcagtgaa
     3601 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     3661 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     3721 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtacttaa gagggggaga
     3781 ccaaagggcg agacgttaag gcctcacgtg acatgtgagc aaaaggccag caaaaggcca
     3841 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3901 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3961 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttacgg
     4021 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4081 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4141 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4201 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4261 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4321 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4381 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4441 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4501 ggaacgaaaa ctcacgttaa gggattttgg tcatgccgtc tcagaagaac tcgtcaagaa
     4561 ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc
     4621 ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct
     4681 gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt
     4741 ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg
     4801 gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt
     4861 ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctctc tcgatgcgat
     4921 gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg
     4981 catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc
     5041 ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagtacag
     5101 ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcttgcagtt
     5161 cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca
     5221 gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata
     5281 gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcataatat
     5341 tattgaagca tttatcaggg ttcgtctcgt cccggtctcc tcccatgcat g
//
