LOCUS       sequence-420046-        4875 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express NES-PUFc-FseI[ADAR2-DD-splitC].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4875)
  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 4875)
  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: pzk491-pmax-NES-PUFc-FseI[ADAR2-DD-splitC]
FEATURES             Location/Qualifiers
     source          1..4875
                     /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(2904..2928)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2981..3022
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3048..3067)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    3077..3158
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     3102..3121
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3356..3944)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3436..3455)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4025..4819)
                     /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(4025..4819)
                     /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(4136..4155)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4746..4765
                     /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 gccggcccca gcagatagac acccaaatcg taaagcaaga ggacagctac
     2221 ggaccaaaat agagtctggt caggggacga ttccagtgcg ctccaatgcg agcatccaaa
     2281 cgtgggacgg ggtgctgcaa ggggagcggc tgctcaccat gtcctgcagt gacaagattg
     2341 cacgctggaa cgtggtgggc atccagggat cactgctcag cattttcgtg gagcccattt
     2401 acttctcgag catcatcctg ggcagccttt accacgggga ccacctttcc agggccatgt
     2461 accagcggat ctccaacata gaggacctgc cacctctcta caccctcaac aagcctttgc
     2521 tcagtggcat cagcaatgca gaagcacggc agccagggaa ggcccccaac ttcagtgtca
     2581 actggacggt aggcgactcc gctattgagg tcatcaacgc cacgactggg aaggatgagc
     2641 tgggccgcgc gtcccgcctg tgtaagcacg cgttgtactg tcgctggatg cgtgtgcacg
     2701 gcaaggttcc ctcccactta ctacgctcca agattaccaa gcccaacgtg taccatgagt
     2761 ccaagctggc ggcaaaggag taccaggccg ccaaggcgcg tctgttcaca gccttcatca
     2821 aggcggggct gggggcctgg gtggagaagc ccaccgagca ggaccagttc tcactcacga
     2881 acgtttagtt aattaagaat tcgacccagc tttcttgtac aaagtggttg atatccagca
     2941 cagtggcggc cgctcgagtc tagagggccc gcggttcgaa ggtaagccta tccctaaccc
     3001 tctcctcggt ctcgattcta cgcgtaccgg ttagcaattg ttttttcgat gagtttggac
     3061 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     3121 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     3181 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     3241 aatgtggtac ttaagagggg gagaccaaag ggcgagacgt taaggcctca cgtgacatgt
     3301 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3361 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3421 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3481 ctgttccgac cctgccgctt acgggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3541 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3601 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3661 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3721 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3781 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3841 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3901 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3961 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatgc
     4021 cgtctcagaa gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg
     4081 cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat
     4141 cacgggtagc caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga
     4201 tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatggg
     4261 tcacgacgag atcctcgccg tcgggcatgc tcgccttgag cctggcgaac agttcggctg
     4321 gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc
     4381 gagtacgtgc tctctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat
     4441 caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa
     4501 ggtgagatga caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg
     4561 cttcagtgac aacgtcgagt acagctgcgc aaggaacgcc cgtcgtggcc agccacgata
     4621 gccgcgctgc ctcgtcttgc agttcattca gggcaccgga caggtcggtc ttgacaaaaa
     4681 gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct
     4741 gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca
     4801 atccatcttg ttcaatcata atattattga agcatttatc agggttcgtc tcgtcccggt
     4861 ctcctcccat gcatg
//
