LOCUS       sequence-420047-        4619 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express NES-PUFc-FseI[ADAR2-DD-splitN].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4619)
  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 4619)
  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: "Nucleic 
            Acids Res."; date: "2023-08-11"; volume: "51(14):e77. doi"; pages: "
            10.1093/nar/gkad547"
COMMENT     Sequence Label: pzk488-pmax-NES-PUFc-FseI[ADAR2-DD-splitN]
FEATURES             Location/Qualifiers
     source          1..4619
                     /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             1011..1037
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     protein_bind    complement(2648..2672)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2725..2766
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2792..2811)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    2821..2902
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     2846..2865
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3100..3688)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3180..3199)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3769..4563)
                     /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(3769..4563)
                     /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(3880..3899)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4490..4509
                     /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 ctgccaccat
     1021 tggaaaggct tacgctgggc ggaggcggat ctgggcgcgc ctctagaggc cgcagccgcc
     1081 ttttggaaga ttttcgaaac aaccggtacc ccaatttaca actgcgggag attgctggac
     1141 atataatgga attttcccaa gaccagcatg ggtccagatt cattcagctg aaactggagc
     1201 gtgccacacc agctgagcgc cagcttgtct tcaatgaaat cctccaggct gcctaccaac
     1261 tcatggtgga tgtgtttggt aattacgtca ttcagaagtt ctttgaattt ggcagtcttg
     1321 aacagaagct ggctttggca gaacggattc gaggccacgt cctgtcattg gcactacaga
     1381 tgtatggcag ccgtgttatc gagaaagctc ttgagtttat tccttcagac cagcagaatg
     1441 agatggttcg ggaactagat ggccatgtct tgaagtgtgt gaaagatcag aatggcaatc
     1501 acgtggttca gaaatgcatt gaatgtgtac agccccagtc tttgcaattt atcatcgacg
     1561 cgtttaaggg acaggtattt gccttatcca cacatcctta tggctgccga gtgattcaga
     1621 gaatcctgga gcactgtctc cctgaccaga cactccctat tttagaggag cttcaccagc
     1681 acacagagca gctggtacag gatcaatatg gaagttatgt aatcgaacat gtactggagc
     1741 acggtcgtcc tgaggataaa agcaaaattg tagcagaaat ccgaggcaat gtacttgtat
     1801 tgagtcagca caaatttgca aacaatgttg tgcagaagtg tgttactcac gcctcacgta
     1861 cggagcgcgc tgtgctcatc gacgaggtgt gcaccatgaa cgacggtccc cacagtgcct
     1921 tatacaccat gatgaaggac cagtatgcca actacgtggt ccagaagatg attgacgtgg
     1981 cggagccagg ccagcggaag atcgtcatgc ataagatccg gccccacatc gcaactcttc
     2041 gtaagtacac ctatggcaag cacattctgg ccaagctgga gaagtactac atgaagaacg
     2101 gtgttgactt agggggcggc cgcgggggag gcggttccgg tggcggcgga agcggaggtg
     2161 gaggatcagg gccggcccag ctgcatttac cgcaggtttt agctgacgct gtctcacgcc
     2221 tggtcctggg taagtttggt gacctgaccg acaacttctc ctcccctcac gctcgcagaa
     2281 aagtgctggc tggagtcgtc atgacaacag gcacagatgt taaagatgcc aaggtgataa
     2341 gtgtttctac aggaacaaaa tgtattaatg gtgaatacat gagtgatcgt ggccttgcat
     2401 taaatgactg ccatgcagaa ataatatctc ggagatcctt gctcagattt ctttatacac
     2461 aacttgagct ttacttaaat aacaaagatg atcaaaaaag atccatcttt cagaaatcag
     2521 agcgaggggg gtttaggctg aaggagaatg tccagtttca tctgtacatc agcacctctc
     2581 cctgtggaga tgccagaatc ttctcaccac atgagccaat cctggaagaa tagttattaa
     2641 gaattcgacc cagctttctt gtacaaagtg gttgatatcc agcacagtgg cggccgctcg
     2701 agtctagagg gcccgcggtt cgaaggtaag cctatcccta accctctcct cggtctcgat
     2761 tctacgcgta ccggttagca attgtttttt cgatgagttt ggacaaacca caactagaat
     2821 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     2881 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     2941 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtacttaaga
     3001 gggggagacc aaagggcgag acgttaaggc ctcacgtgac atgtgagcaa aaggccagca
     3061 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3121 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3181 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3241 gcttacggga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3301 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3361 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3421 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3481 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3541 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3601 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3661 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3721 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgccgtctc agaagaactc
     3781 gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac
     3841 gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc
     3901 tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg
     3961 gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc
     4021 gccgtcgggc atgctcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg
     4081 ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac gtgctctctc
     4141 gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg
     4201 ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag
     4261 atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc
     4321 gagtacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc
     4381 ttgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg
     4441 cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata
     4501 gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat
     4561 cataatatta ttgaagcatt tatcagggtt cgtctcgtcc cggtctcctc ccatgcatg
//
