LOCUS       sequence-273378-        4065 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human codon-optimized AcrIIA6 D1024 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 4065)
  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 4065)
  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_AcrIIA6_D1024
FEATURES             Location/Qualifiers
     source          1..4065
                     /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"
     primer_bind     complement(1886..1903)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1892..2116
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             2289..3083
                     /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             2289..3083
                     /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(2343..2362)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2953..2972
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      3408..3996
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3897..3916
                     /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 gccatgaaga tcaacgacga cataaaggaa
     1321 ttgatcctcg aatacatgtc ccgatatttt aaattcgaga atgacttcta taaactgccg
     1381 ggaatcaaat ttaccgatgc taactggcag aagttcaaaa atggggggac tgatatcgag
     1441 aagatggggg cggccagggt gaatgcgatg ctttcatgtc tcttcgaaga cttcgagttg
     1501 gcaatgattg gaaaagccca aactaactat tatattgaca attctctcaa gcttaatatg
     1561 ccattttacg cttattacga catgtttaaa aagcagttgc ttattaactg gctgaaaaat
     1621 aatcgcgacg acgttatttg cggcacgggt aggatgtata ctgcgtcagg gaactatatt
     1681 gcaaacgcgt acctggaggt ggctctggag tctagccgac tgggcggtgg agagtatatg
     1741 cttcagatgc gcttcaaaaa ttatagtcgc agtcaagagc ctatcccatc aggacgacag
     1801 aaccgacttg aatggatcga aaacaatctt gagaacatcc ggtgataact cgagtctaga
     1861 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     1921 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     1981 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2041 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     2101 gcggtgggct ctatggcttc tactgggcgg ttttatggac agcaagcgaa ccggaattgc
     2161 cagctggggc gccctctggt aaggttggga agccctgcaa agtaaactgg atggctttct
     2221 tgccgccaag gatctgatgg cgcaggggat caagctctga tcaagagaca ggatgaggat
     2281 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     2341 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     2401 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     2461 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     2521 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     2581 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     2641 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     2701 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     2761 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     2821 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     2881 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     2941 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     3001 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     3061 gccttcttga cgagttcttc tgaattatta acgcttacaa tttcctgatg cggtattttc
     3121 tccttacgca tctgtgcggt atttcacacc gcatcaggtg gcacttttcg gggaaatgtg
     3181 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3241 caataaccct gataaatgct tcaataatag cacgtgctaa aacttcattt ttaatttaaa
     3301 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     3361 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3421 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3481 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3541 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3601 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3661 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3721 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3781 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3841 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3901 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3961 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4021 cggttcctgg ccttttgctg gccttttgct cacatgttct tgctg
//
