LOCUS       sequence-226954-        4551 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for expression of SV40NLS-AcrIIA4 in mammalian cells (CMV
            promoter driven).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4551)
  AUTHORS   Bubeck F, Hoffmann MD, Harteveld Z, Aschenbrenner S, Bietz A, 
            Waldhauer MC, Borner K, Fakhiri J, Schmelas C, Dietz L, Grimm D, 
            Correia BE, Eils R, Niopek D
  TITLE     Engineered anti-CRISPR proteins for optogenetic control of 
            CRISPR-Cas9.
  JOURNAL   Nat Methods. 2018 Oct 30. pii: 10.1038/s41592-018-0178-9. doi: 
            10.1038/s41592-018-0178-9.
  PUBMED    30377362
REFERENCE   2  (bases 1 to 4551)
  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 
            Methods. 2018 Oct 30. pii: 10.1038/s41592-018-0178-9. doi: 
            10.1038/s41592-018-0178-9."
COMMENT     Sequence Label: pAAV-CMV-NLS-AcrIIA4
FEATURES             Location/Qualifiers
     source          1..4551
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(28..456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(138..159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     350..369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     583..605
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     597..614
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     598..614
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        624..642
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   699..828
                     /label=AAV2 ITR
     enhancer        848..1227
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1228..1431
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1382..1402
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        1476..1494
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1520..1540
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1923..1940)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1929..2153
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2363..2383)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2363..2381)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2402..2418)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2402..2418)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2415..2437)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2426..2442
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2450..2456,2457..2474,2475..2480))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2495..2516
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2804..3392)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2884..2903)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3563..4354,4355..4423))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4186..4205
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4424..4528)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
       61 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
      121 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
      181 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
      241 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
      301 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
      361 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
      421 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tgssccattc aggctgcgca
      481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
      541 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
      601 aaacgacggc cagtgagcgc gcgtaatacg actcactata gggcgaattg gagctctcta
      661 gaatgcaggg gggggggggg gggkkgscca ctccctctct gcgcgctcgc tcgctcactg
      721 aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg
      781 agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctca gatctgatat
      841 cgtcgacgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
      901 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      961 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     1021 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     1081 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     1141 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     1201 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     1261 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     1321 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     1381 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta
     1441 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc
     1501 caagctggct agcatggccc cgaagaaaaa gcgaaaggtg ggcggaagcg gaggttccat
     1561 gaacattaac gacctgattc gggaaataaa gaacaaggac tacacggtca agctgtcagg
     1621 caccgactcc aactctataa cacagctcat tataagggtt aataatgatg gcaacgaata
     1681 tgtcatctct gaatccgaga atgaatcaat cgtggagaaa ttcatcagtg catttaaaaa
     1741 cggctggaac caggagtacg aagatgagga agagttctac aatgatatgc aaactatcac
     1801 actgaaatct gagctgaact agctcgagcg gccgccactg tgctggatat ctgcagaatt
     1861 ccaccacact ggactagtgg atccgagctc ggtaccaagc ttaagtttaa accgctgatc
     1921 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     1981 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     2041 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     2101 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggggcgcgc
     2161 cgatatcaga tctgggaaac cagatgatgg aggtacccac tccctctatg cgcgctcgct
     2221 cactcactcg gccctggaga cctttggtct ccgccctgga gacctttggt ctccagggcc
     2281 gagtgagtga gcgagcgcgc atagagggag tgggtaggac gcgtcctgca ggatgcatac
     2341 tagtggtacc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
     2401 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     2461 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     2521 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     2581 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     2641 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     2701 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     2761 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     2821 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2881 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2941 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3001 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3061 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3121 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3181 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3241 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3301 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3361 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3421 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     3481 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     3541 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     3601 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     3661 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     3721 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     3781 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     3841 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     3901 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     3961 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4021 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4081 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4141 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     4201 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     4261 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     4321 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     4381 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     4441 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     4501 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca c
//
