LOCUS       sequence-273371-        3936 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 3936)
  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 3936)
  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..3936
                     /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"
     regulatory      1289..1298
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1758..1775)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1764..1988
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             2161..2955
                     /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             2161..2955
                     /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(2215..2234)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2825..2844
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      3280..3868
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3769..3788
                     /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 cgccatggcc tatggtaaaa gtaggtataa
     1321 ctcctatcgc aaacgctctt tcaacaggtc taacaaacaa aggagagaat acgcacagga
     1381 aatggaccgc cttgaaaagg catttgagaa cctcgacgga tggtacttga gtagcatgaa
     1441 agattctgcg tataaggatt tcggtaagta cgaaatccga ctttcaaatc actcagccga
     1501 caataaatat catgacctgg agaacggccg cttgatcgtg aatatcaaag caagcaaact
     1561 taactttgtc gatattatcg agaataaact cgacaagatc atcgaaaaaa ttgataagct
     1621 ggaccttgat aaatatcgct tcatcaatgc gaccaacctg gagcacgaca ttaagtgcta
     1681 ctataaaggg tttaagacta agaaagaggt tatctgataa ctcgagtcta gagggcccgt
     1741 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     1801 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     1861 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     1921 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     1981 ctctatggct tctactgggc ggttttatgg acagcaagcg aaccggaatt gccagctggg
     2041 gcgccctctg gtaaggttgg gaagccctgc aaagtaaact ggatggcttt cttgccgcca
     2101 aggatctgat ggcgcagggg atcaagctct gatcaagaga caggatgagg atcgtttcgc
     2161 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     2221 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     2281 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     2341 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     2401 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     2461 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     2521 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     2581 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     2641 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     2701 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     2761 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     2821 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     2881 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     2941 gacgagttct tctgaattat taacgcttac aatttcctga tgcggtattt tctccttacg
     3001 catctgtgcg gtatttcaca ccgcatcagg tggcactttt cggggaaatg tgcgcggaac
     3061 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3121 ctgataaatg cttcaataat agcacgtgct aaaacttcat ttttaattta aaaggatcta
     3181 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     3241 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3301 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3361 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3421 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3481 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3541 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3601 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3661 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3721 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3781 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3841 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3901 ggccttttgc tggccttttg ctcacatgtt cttgct
//
