LOCUS       sequence-419900-        5418 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express PUFc-RESCUE-S.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5418)
  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 5418)
  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: pzk384-pmax-PUFc-FseI-RESCUE-SalI-NES
FEATURES             Location/Qualifiers
     source          1..5418
                     /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             3402..3428
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     protein_bind    complement(3447..3471)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3524..3565
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3591..3610)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    3620..3701
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     3645..3664
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3899..4487)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3979..3998)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4568..5362)
                     /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(4568..5362)
                     /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(4679..4698)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5289..5308
                     /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 ccggccctcc aaaaaaagtt ggaggaatta gaactaggat
     2221 cccagctgca tttaccgcag gttttagctg acgctgtctc acgcctggtc ataggtaagt
     2281 ttggtgacct gaccgacaac ttctcctccc ctcacgctcg cagaataggt ctggctggag
     2341 tcgtcatgac aacaggcaca gatgttaaag atgccaaggt gatatgtgtt tctacaggag
     2401 caaaatgtat taatggtgaa tacctaagtg atcgtggcct tgcattaaat gactgccatg
     2461 cagaaatagt atctcggaga tccttgctca gatttcttta tacacaactt gagctttact
     2521 taaataacga ggatgatcaa aaaagatcca tctttcagaa atcagagcga ggggggttta
     2581 ggctgaagga gaatatacag tttcatctgt acatcagcac ctctccctgt ggagatgcca
     2641 gaatcttctc accacatgag gcaatcctgg aagaaccagc agatagacac ccaaatcgta
     2701 aagcaagagg acagctacgg accaaaatag aggctggtca ggggacgatt ccagtgcgca
     2761 acaatgcgag catccaaacg tgggacgggg tgctgcaagg ggagcggctg ctcaccatgt
     2821 cctgcagtga caagattgca cgctggaacg tggtgggcat ccagggatca ctgctcagca
     2881 ttttcgtgga gcccatttac ttctcgagca tcatcctggg cagcctttac cacggggacc
     2941 acctttccag ggccatgtac cagcggatct ccaacataga ggacctgcca cctctctaca
     3001 ccctcaacaa gcctttgctc acaggcatca gcaatgcaga agcacggcag ccagggaagg
     3061 cccccatatt cagtgtcaac tggacggtag gcgactccgc tattgaggtc atcaacgcca
     3121 cgactgggaa gggagagctg ggccgcgcgt cccgcctgtg taagcacgcg ttgtactgtc
     3181 gctggatgcg tgtgcacggc aaggttccct cccacttact acgctccaag attaccaagc
     3241 ccaacgtgta ccatgagaca aagctggcgg caaaggagta ccaggccgcc aaggcgcgtc
     3301 tgttcacagc cttcatcaag gcggggctgg gggcctgggt ggagaagccc accgagcagg
     3361 accagttctc actcacggtc gacgggagcg gatccggctc tctgccacca ttggaaaggc
     3421 ttacgctgta gttaattaag aattcgaccc agctttcttg tacaaagtgg ttgatatcca
     3481 gcacagtggc ggccgctcga gtctagaggg cccgcggttc gaaggtaagc ctatccctaa
     3541 ccctctcctc ggtctcgatt ctacgcgtac cggttagcaa ttgttttttc gatgagtttg
     3601 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     3661 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     3721 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
     3781 acaaatgtgg tacttaagag ggggagacca aagggcgaga cgttaaggcc tcacgtgaca
     3841 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3901 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3961 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4021 ctcctgttcc gaccctgccg cttacgggat acctgtccgc ctttctccct tcgggaagcg
     4081 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4141 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4201 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4261 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4321 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4381 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4441 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4501 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4561 tgccgtctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag
     4621 cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa
     4681 tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt
     4741 cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat
     4801 gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg
     4861 ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca
     4921 tccgagtacg tgctctctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg
     4981 gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag
     5041 caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc
     5101 ccgcttcagt gacaacgtcg agtacagctg cgcaaggaac gcccgtcgtg gccagccacg
     5161 atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa
     5221 aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg
     5281 tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt
     5341 gcaatccatc ttgttcaatc ataatattat tgaagcattt atcagggttc gtctcgtccc
     5401 ggtctcctcc catgcatg
//
