LOCUS       sequence-263829-        3940 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human codon-optimized AcrIIA5 D4276 in mammalian cells. 
            ORF flanked by NotI and XhoI restriction sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3940)
  AUTHORS   Hynes AP, Rousseau GM, Agudelo D, Goulet A, Amigues B, Loehr J, 
            Romero DA, Fremaux C, Horvath P, Doyon Y, Cambillau C, Moineau S
  TITLE     Widespread anti-CRISPR proteins in virulent bacteriophages inhibit a
            range of Cas9 proteins.
  JOURNAL   Nat Commun. 2018 Jul 25;9(1):2919. doi: 10.1038/s41467-018-05092-w.
  PUBMED    30046034
REFERENCE   2  (bases 1 to 3940)
  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: "Nat 
            Commun."; date: "2018-07-25"; volume: "9(1):2919. doi"; pages: " 
            10.1038/s41467-018-05092-w"
COMMENT     Sequence Label: pVAX_CBI_AcrIIA5_D4276
FEATURES             Location/Qualifiers
     source          1..3940
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        100..403
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        404..606
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     557..577
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     603..627
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          755..1133
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     CDS             1220..1267
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     regulatory      1295..1304
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1764..1781)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1770..1994
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             2167..2961
                     /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             2167..2961
                     /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(2221..2240)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2831..2850
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      3287..3875
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3776..3795
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gactcttcgc gatgtacggg ccagatatac gcgtggagct agttattaat agtaatcaat
       61 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
      121 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      181 tcccatagta acgtcaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      241 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      301 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      361 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      421 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      481 tgacgtcaat gggagtttgt tttgcaccaa aatcaacggg actttccaaa atgtcgtaac
      541 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      601 agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc
      661 catagaagac accgggaccg atccagcctc cgcggattcg aatcccggcc gggaacggtg
      721 cattggaacg cggattcccc gtgccaagag tgacgtaagt accgcctata gagtctatag
      781 gcccacaaaa aatgctttct tcttttaata tacttttttg tttatcttat ttctaatact
      841 ttccctaatc tctttctttc agggcaataa tgatacaatg tatcatgcct ctttgcacca
      901 ttctaaagaa taacagtgat aatttctggg ttaaggcaat agcaatattt ctgcatataa
      961 atatttctgc atataaattg taactgatgt aagaggtttc atattgctaa tagcagctac
     1021 aatccagcta ccattctgct tttattttat ggttgggata aggctggatt attctgagtc
     1081 caagctaggc ccttttgcta atcatgttca tacctcttat cttcctccca cagctcctgg
     1141 gcaacgtgct ggtctgtgtg ctggcccatc actttggcaa agaattggga ttcgaacatc
     1201 gattgaattc ctgtacaaga aaaggccggc ggccacgaaa aaggccggcc aggcaaaaaa
     1261 gaaaaagggc ggaggctctg gcggcggaag cgcggccgcc atggcctatg gtaaaagtag
     1321 gtataactcc tatcgcaaac gctctttcaa caggtctaac aaacaaagga gagaatacgc
     1381 acaggaaatg gaccgccttg aaaaggcatt tgagaacctc gacggatggt acttgagtag
     1441 catgaaagat tctgcgtata aggatttcgg taagtacgaa atccgacttt caaatcactc
     1501 agccgacaat aaatatcatg acctggagaa cggccgcttg atcgtgaata tcaaagcaag
     1561 caaacttaac tttgtcgata ttatcgagaa taaactcgac aagatcatcg aaaaaattga
     1621 taagctggac cttgataaat atcgcttcat caatgcgacc aacctggagc acgacattaa
     1681 gtgctactat aaagggttta agactaagaa agaggttatc tgataactcg agtctagagg
     1741 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     1801 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     1861 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     1921 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     1981 ggtgggctct atggcttcta ctgggcggtt ttatggacag caagcgaacc ggaattgcca
     2041 gctggggcgc cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttctcg
     2101 ccgccaagga tctgatggcg caggggatca agctctgatc aagagacagg atgaggatcg
     2161 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     2221 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     2281 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     2341 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     2401 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     2461 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     2521 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     2581 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     2641 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     2701 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     2761 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     2821 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     2881 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     2941 cttcttgacg agttcttctg aattattaac gcttacaatt tcctgatgcg gtattttctc
     3001 cttacgcatc tgtgcggtat ttcacaccgc atacaggtgg cacttttcgg ggaaatgtgc
     3061 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3121 aataaccctg ataaatgctt caataatagc acgtgctaaa acttcatttt taatttaaaa
     3181 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     3241 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3301 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3361 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3421 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3481 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3541 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3601 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3661 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3721 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3781 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3841 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3901 ggttcctggg cttttgctgg ccttttgctc acatgttctt
//
