LOCUS       sequence-232763-        5191 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for expression of AcrIIC3 (no miR binding sites, control 
            vector).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5191)
  AUTHORS   Hoffmann MD, Aschenbrenner S, Grosse S, Rapti K, Domenger C, Fakhiri
            J, Mastel M, Borner K, Eils R, Grimm D, Niopek D
  TITLE     Cell-specific CRISPR-Cas9 activation by microRNA-dependent 
            expression of anti-CRISPR proteins.
  JOURNAL   Nucleic Acids Res. 2019 Apr 15. pii: 5458120. doi: 
            10.1093/nar/gkz271.
  PUBMED    30982889
REFERENCE   2  (bases 1 to 5191)
  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: "Nucleic 
            Acids Res. 2019 Apr 15. pii: 5458120. doi: 10.1093/nar/gkz271."
COMMENT     Sequence Label: AAV CMV-driven AcrIIC3-scaffold (2xBsmBI sites)
FEATURES             Location/Qualifiers
     source          1..5191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     enhancer        370..749
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        750..953
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     904..924
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        998..1016
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1036..1045
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1512..1529)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1518..1742
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(1948..1966)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1976..1993)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1976..1992)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1985..2007)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(2133..2588)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2270..2291)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2482..2501
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2612..2716
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2717..2785,2786..3577)
                     /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     complement(2935..2954)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3748..4336
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4237..4256
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    4624..4645
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4660..4665,4666..4683,4684..4690)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4698..4714
                     /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)."
     primer_bind     4703..4725
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4722..4738
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4722..4738
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4757..4777
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4759..4777
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg agtcttcgag gaagacaggt ttaagagcta tgctggaaac agcatagcaa
      301 gtttaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt
      361 tgcggccgcg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
      421 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      481 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      541 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      601 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      661 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      721 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      781 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      841 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      901 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
      961 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
     1021 cccaagctgg ctagcgccac catggctttc aagagagcta ttatatttac gtcctttaat
     1081 ggttttgaga aggtctctcg aacagagaag cgacgccttg ccaagataat taacgcgagg
     1141 gtctcaatta ttgatgagta tctcagagcc aaggatacaa acgcgagcct ggatggccaa
     1201 taccgagcct ttctgtttaa cgacgaaagc cccgcgatga cagagttcct cgcaaaactg
     1261 aaagcattcg cggagagctg cacgggaata tcaatagatg cgtgggagat tgaagagagc
     1321 gagtatgttc gactgcccgt tgaacgccgg gatttcctgg ccgctgcgaa tggcaaggaa
     1381 atcttcaaga tataactcga gcggccgcca ctgtgctgga tatctgcaga attagagacg
     1441 ccgttgatgt tgaagtttat ataccgttga tgttgaagtc gtctctagct taagtttaaa
     1501 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     1561 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     1621 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     1681 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     1741 gggtcgacga tatcagatct gaggaacccc tagtgatgga gttggccact ccctctctgc
     1801 gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg acctttggtc
     1861 gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggcccccccc cccccccccc
     1921 cctgcattct agagagctcc aattcgccct atagtgagtc gtattacgcg cgctcactgg
     1981 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     2041 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     2101 cccaacagtt gcgcagcctg aatggcgaat ggaaattgta agcgttaata ttttgttaaa
     2161 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     2221 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     2281 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     2341 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     2401 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     2461 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     2521 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     2581 gggcgcgtgt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     2641 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     2701 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     2761 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     2821 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     2881 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     2941 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     3001 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     3061 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     3121 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     3181 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     3241 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     3301 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     3361 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     3421 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     3481 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     3541 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     3601 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     3661 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3721 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3781 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3841 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3901 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3961 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4021 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4081 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4141 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4201 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4261 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4321 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4381 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     4441 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     4501 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     4561 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     4621 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     4681 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     4741 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg gtaccactag
     4801 tatgcatcct gcaggacgcg tcctacccac tccctctatg cgcgctcgct cactcactcg
     4861 gccctggaga ccaaaggtct ccagggccga gtgagtgagc gagcgcgcat agagggagtg
     4921 ggtacctcca tcatctggtt tcccagatct gatatcggcg cgcccgagtc caacacccgt
     4981 gggaatccca tgggcaccat ggcccctcgc tccaaaaatg ctttcgcgtc gcgcagacac
     5041 tgctcggtag tttcggggat cagcgtttga gtaagagccc gcgtctgaac cctccgcgcc
     5101 gccccggccc cagtggaaag acgcgcaggc aaaacgcacc acgtgacgga gcgtgaccgc
     5161 gcgccgagcg cgcgccaagg tcgggcagga a
//
