LOCUS       sequence-273367-        3777 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human codon-optimized AcrIIA4 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 3777)
  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 3777)
  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_AcrIIA4
FEATURES             Location/Qualifiers
     source          1..3777
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        93..396
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        397..600
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     551..571
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     597..621
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          749..1127
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     CDS             1214..1261
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(1599..1616)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1605..1829
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             2002..2796
                     /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             2002..2796
                     /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(2056..2075)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2666..2685
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      3121..3709
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3610..3629
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcttcgcgat gtacgggcca gatatacgcg tctagttatt aatagtaatc aattacgggg
       61 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      121 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      181 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      241 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      301 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      361 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      421 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      481 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      541 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      601 cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga
      661 agacaccggg accgatccag cctccgcgga ttcgaatccc ggccgggaac ggtgcattgg
      721 aacgcggatt ccccgtgcca agagtgacgt aagtaccgcc tatagagtct ataggcccac
      781 aaaaaatgct ttcttctttt aatatacttt tttgtttatc ttatttctaa tactttccct
      841 aatctctttc tttcagggca ataatgatac aatgtatcat gcctctttgc accattctaa
      901 agaataacag tgataatttc tgggttaagg caatagcaat atttctgcat ataaatattt
      961 ctgcatataa attgtaactg atgtaagagg tttcatattg ctaatagcag ctacaatcca
     1021 gctaccattc tgcttttatt ttatggttgg gataaggctg gattattctg agtccaagct
     1081 aggccctttt gctaatcatg ttcatacctc ttatcttcct cccacagctc ctgggcaacg
     1141 tgctggtctg tgtgctggcc catcactttg gcaaagaatt gggattcgaa catcgattga
     1201 attcctgtac aagaaaaggc cggcggccac gaaaaaggcc ggccaggcaa aaaagaaaaa
     1261 gggcggaggc tctggcggcg gaagcgcggc cgccatgaac attaacgacc tcatacgaga
     1321 gattaagaac aaagattaca ccgtcaaact gtcaggaact gatagtaact caatcaccca
     1381 gcttattatc agggtaaaca atgatgggaa tgaatatgtg atatctgaga gcgaaaacga
     1441 gtctatcgtc gagaaattca tttccgcttt taagaacggg tggaatcagg aatatgagga
     1501 tgaagaagaa ttttacaatg acatgcagac gatcacgttg aaaagtgaac tgaactgata
     1561 actcgagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     1621 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     1681 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     1741 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     1801 gcatgctggg gatgcggtgg gctctatggc ttctactggg cggttttatg gacagcaagc
     1861 gaaccggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg caaagtaaac
     1921 tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcaagctc tgatcaagag
     1981 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     2041 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     2101 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     2161 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     2221 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     2281 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     2341 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     2401 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     2461 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     2521 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     2581 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     2641 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     2701 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     2761 atcgccttct atcgccttct tgacgagttc ttctgaatta ttaacgctta caatttcctg
     2821 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatcag gtggcacttt
     2881 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     2941 tccgctcatg agacaataac cctgataaat gcttcaataa tagcacgtgc taaaacttca
     3001 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     3061 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3121 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3181 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3241 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3301 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3361 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3421 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3481 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3541 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3601 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3661 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3721 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tcttgct
//
