LOCUS       sequence-420052-        4734 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express MCP_NES-ADARdd(E488Q).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4734)
  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 4734)
  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: pmax-MCP_NES-ADARdd(E488Q)
FEATURES             Location/Qualifiers
     source          1..4734
                     /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             1074..1460
                     /codon_start=1
                     /product="bacteriophage MS2 coat protein"
                     /label=MS2-N55K
                     /note="binds with high affinity to a specific stem-loop 
                     structure in the viral RNA (Lim et al., 1994)"
                     /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT
                     CSVRQSSAQKRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI
                     VKAMQGLLKDGNPIPSAIAANSGIY"
     CDS             1536..1562
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     protein_bind    complement(2763..2787)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2840..2881
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2907..2926)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    2936..3017
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     2961..2980
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3215..3803)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3295..3314)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3884..4678)
                     /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(3884..4678)
                     /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(3995..4014)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4605..4624
                     /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 atggcttcaa
     1081 actttactca gttcgtgctc gtggacaatg gtgggacagg ggatgtgaca gtggctcctt
     1141 ctaatttcgc taatggggtg gcagagtgga tcagctccaa ctcacggagc caggcctaca
     1201 aggtgacatg cagcgtcagg cagtctagtg cccagaagag aaagtatacc atcaaggtgg
     1261 aggtccccaa agtggctacc cagacagtgg gcggagtcga actgcctgtc gccgcttgga
     1321 ggtcctacct gaacatggag ctcactatcc caattttcgc taccaattct gactgtgaac
     1381 tcatcgtgaa ggcaatgcag gggctcctca aagacggtaa tcctatccct tccgccatcg
     1441 ccgctaactc aggtatctac agcgctggcg gccgcggggg aggcggttcc ggtggcggcg
     1501 gaagcggagg tggaggatca gggccggccc tgcagctgcc tccacttgaa agactgacac
     1561 tgggatccgg aggaggtgga agccagctgc atttaccgca ggttttagct gacgctgtct
     1621 cacgcctggt cctgggtaag tttggtgacc tgaccgacaa cttctcctcc cctcacgctc
     1681 gcagaaaagt gctggctgga gtcgtcatga caacaggcac agatgttaaa gatgccaagg
     1741 tgataagtgt ttctacagga acaaaatgta ttaatggtga atacatgagt gatcgtggcc
     1801 ttgcattaaa tgactgccat gcagaaataa tatctcggag atccttgctc agatttcttt
     1861 atacacaact tgagctttac ttaaataaca aagatgatca aaaaagatcc atctttcaga
     1921 aatcagagcg aggggggttt aggctgaagg agaatgtcca gtttcatctg tacatcagca
     1981 cctctccctg tggagatgcc agaatcttct caccacatga gccaatcctg gaagaaccag
     2041 cagatagaca cccaaatcgt aaagcaagag gacagctacg gaccaaaata gagtctggtc
     2101 aggggacgat tccagtgcgc tccaatgcga gcatccaaac gtgggacggg gtgctgcaag
     2161 gggagcggct gctcaccatg tcctgcagtg acaagattgc acgctggaac gtggtgggca
     2221 tccagggatc actgctcagc attttcgtgg agcccattta cttctcgagc atcatcctgg
     2281 gcagccttta ccacggggac cacctttcca gggccatgta ccagcggatc tccaacatag
     2341 aggacctgcc acctctctac accctcaaca agcctttgct cagtggcatc agcaatgcag
     2401 aagcacggca gccagggaag gcccccaact tcagtgtcaa ctggacggta ggcgactccg
     2461 ctattgaggt catcaacgcc acgactggga aggatgagct gggccgcgcg tcccgcctgt
     2521 gtaagcacgc gttgtactgt cgctggatgc gtgtgcacgg caaggttccc tcccacttac
     2581 tacgctccaa gattaccaag cccaacgtgt accatgagtc caagctggcg gcaaaggagt
     2641 accaggccgc caaggcgcgt ctgttcacag ccttcatcaa ggcggggctg ggggcctggg
     2701 tggagaagcc caccgagcag gaccagttct cactcacgaa cgtttgatta attaagaatt
     2761 cgacccagct ttcttgtaca aagtggttga tatccagcac agtggcggcc gctcgagtct
     2821 agagggcccg cggttcgaag gtaagcctat ccctaaccct ctcctcggtc tcgattctac
     2881 gcgtaccggt tagcaattgt tttttcgatg agtttggaca aaccacaact agaatgcagt
     2941 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     3001 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     3061 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtact taagaggggg
     3121 agaccaaagg gcgagacgtt aaggcctcac gtgacatgtg agcaaaaggc cagcaaaagg
     3181 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3241 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3301 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3361 cgggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3421 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3481 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3541 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3601 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3661 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3721 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3781 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3841 agtggaacga aaactcacgt taagggattt tggtcatgcc gtctcagaag aactcgtcaa
     3901 gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga
     3961 agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt
     4021 cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat
     4081 tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt
     4141 cgggcatgct cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt
     4201 cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct ctctcgatgc
     4261 gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca
     4321 ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct
     4381 gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagta
     4441 cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcttgca
     4501 gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg
     4561 acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga
     4621 atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcataa
     4681 tattattgaa gcatttatca gggttcgtct cgtcccggtc tcctcccatg catg
//
