LOCUS       sequence-273234-        3885 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses human codon-optimized AcrIIA2 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 3885)
  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 3885)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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_AcrIIA2
FEATURES             Location/Qualifiers
     source          1..3885
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        95..398
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        399..602
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     553..573
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     599..623
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          751..1129
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     CDS             1216..1263
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(1709..1726)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1715..1939
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             2112..2906
                     /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             2112..2906
                     /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(2166..2185)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2776..2795
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      3231..3819
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3720..3739
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgcttcgcg atgtacgggc cagatatacg cgtctagtta ttaatagtaa tcaattacgg
       61 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      121 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      181 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      241 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      301 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      361 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      421 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      481 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      541 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      601 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
      661 gaagacaccg ggaccgatcc agcctccgcg gattcgaatc ccggccggga acggtgcatt
      721 ggaacgcgga ttccccgtgc caagagtgac gtaagtaccg cctatagagt ctataggccc
      781 acaaaaaatg ctttcttctt ttaatatact tttttgttta tcttatttct aatactttcc
      841 ctaatctctt tctttcaggg caataatgat acaatgtatc atgcctcttt gcaccattct
      901 aaagaataac agtgataatt tctgggttaa ggcaatagca atatttctgc atataaatat
      961 ttctgcatat aaattgtaac tgatgtaaga ggtttcatat tgctaatagc agctacaatc
     1021 cagctaccat tctgctttta ttttatggtt gggataaggc tggattattc tgagtccaag
     1081 ctaggccctt ttgctaatca tgttcatacc tcttatcttc ctcccacagc tcctgggcaa
     1141 cgtgctggtc tgtgtgctgg cccatcactt tggcaaagaa ttgggattcg aacatcgatt
     1201 gaattcctgt acaagaaaag gccggcggcc acgaaaaagg ccggccaggc aaaaaagaaa
     1261 aagggcggag gctctggcgg cggaagcgcg gccgccatga cgctgacccg cgctcagaaa
     1321 aaatacgccg aggcgatgca tgagtttatc aatatggttg atgactttga agaatcaacg
     1381 cctgactttg caaaagaggt tctgcacgac tccgactatg tggtcattac aaaaaacgag
     1441 aaatatgccg tggcactctg tagtctctcc acagatgaat gtgagtacga tactaacttg
     1501 tatttggatg aaaagctcgt cgattacagc acagttgatg tcaacggagt gacatattac
     1561 atcaatatag tggaaacaaa tgacatagat gatcttgaaa ttgcgaccga cgaggacgag
     1621 atgaagtctg gaaaccaaga gattattctt aagtccgaac tgaagtgata actcgagtct
     1681 agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     1741 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     1801 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     1861 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     1921 gatgcggtgg gctctatggc ttctactggg cggttttatg gacagcaagc gaaccggaat
     1981 tgccagctgg ggcgccctct ggtaaggttg ggaagccctg caaagtaaac tggatggctt
     2041 tcttgccgcc aaggatctga tggcgcaggg gatcaagctc tgatcaagag acaggatgag
     2101 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     2161 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     2221 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     2281 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     2341 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     2401 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     2461 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     2521 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     2581 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     2641 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     2701 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     2761 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     2821 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     2881 atcgccttct tgacgagttc ttctgaatta ttaacgctta caatttcctg atgcggtatt
     2941 ttctccttac gcatctgtgc ggtatttcac accgcatcag gtggcacttt tcggggaaat
     3001 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3061 agacaataac cctgataaat gcttcaataa tagcacgtgc taaaacttca tttttaattt
     3121 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3181 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3241 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3301 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3361 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     3421 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3481 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3541 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3601 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3661 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3721 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3781 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3841 ttacggttcc tggccttttg ctggcctttt gctcacatgt tcttg
//
