LOCUS       sequence-273369-        3936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human codon-optimized AcrIIA5 D1126 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_D1126
FEATURES             Location/Qualifiers
     source          1..3936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        92..395
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        396..599
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     550..570
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     596..620
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          748..1126
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     CDS             1213..1260
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     regulatory      1288..1297
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1757..1774)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1763..1987
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             2160..2954
                     /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             2160..2954
                     /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(2214..2233)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2824..2843
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      3279..3867
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3768..3787
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cttcgcgatg tacgggccag atatacgcgt ctagttatta atagtaatca attacggggt
       61 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
      121 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      181 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
      241 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
      301 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
      361 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
      421 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
      481 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
      541 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
      601 gtttagtgaa ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa
      661 gacaccggga ccgatccagc ctccgcggat tcgaatcccg gccgggaacg gtgcattgga
      721 acgcggattc cccgtgccaa gagtgacgta agtaccgcct atagagtcta taggcccaca
      781 aaaaatgctt tcttctttta atatactttt ttgtttatct tatttctaat actttcccta
      841 atctctttct ttcagggcaa taatgataca atgtatcatg cctctttgca ccattctaaa
      901 gaataacagt gataatttct gggttaaggc aatagcaata tttctgcata taaatatttc
      961 tgcatataaa ttgtaactga tgtaagaggt ttcatattgc taatagcagc tacaatccag
     1021 ctaccattct gcttttattt tatggttggg ataaggctgg attattctga gtccaagcta
     1081 ggcccttttg ctaatcatgt tcatacctct tatcttcctc ccacagctcc tgggcaacgt
     1141 gctggtctgt gtgctggccc atcactttgg caaagaattg ggattcgaac atcgattgaa
     1201 ttcctgtaca agaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa aaagaaaaag
     1261 ggcggaggct ctggcggcgg aagcgcggcc gccatggcct atggtaaaag taggtataac
     1321 tcctatcgca aacgcaactt cagcatctct gacaatcaaa ggagagaata cgcaaagaaa
     1381 atgaaggaac ttgaacaggc atttgagaac ctcgacggat ggtacttgag tagcatgaaa
     1441 gattctgcgt ataaggattt cggtaagtac gaaatccgac tttcaaatca ctcagccgac
     1501 aatcgctatc atgacctgga gaacggccgc ttgatcgtga atgtgaaagc aagcaaactt
     1561 aactttgtcg atattatcga gaataaactc ggcaagatca tcgaaaaaat tgataccctg
     1621 gaccttgata aatatcgctt catcaatgcg accaagctgg agagggacat taagtgctac
     1681 tataaagggt ataagactaa gaaagacgtt atctgataac tcgagtctag agggcccgtt
     1741 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     1801 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     1861 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     1921 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     1981 tctatggctt ctactgggcg gttttatgga cagcaagcga accggaattg ccagctgggg
     2041 cgccctctgg taaggttggg aagccctgca aagtaaactg gatggctttc ttgccgccaa
     2101 ggatctgatg gcgcagggga tcaagctctg atcaagagac aggatgagga tcgtttcgca
     2161 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     2221 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     2281 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     2341 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     2401 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     2461 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     2521 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     2581 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     2641 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     2701 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     2761 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     2821 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     2881 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     2941 acgagttctt ctgaattatt aacgcttaca atttcctgat gcggtatttt ctccttacgc
     3001 atctgtgcgg tatttcacac cgcatcaggt ggcacttttc ggggaaatgt gcgcggaacc
     3061 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3121 tgataaatgc ttcaataata gcacgtgcta aaacttcatt tttaatttaa aaggatctag
     3181 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     3241 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     3301 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     3361 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     3421 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     3481 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     3541 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     3601 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     3661 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     3721 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     3781 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     3841 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     3901 gccttttgct ggccttttgc tcacatgttc ttgctg
//
