LOCUS       Exported                5122 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  AAV vector for expression of AcrIIC1 with two miR-122 binding sites.
ACCESSION   .
VERSION     .
KEYWORDS    AAV CMV-driven AcrIIC1-2xmiR-122 target sites
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5122)
  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 5122)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5122
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..241
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     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
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     complement(1440..1457)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1446..1670
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1690..1819
                     /label=AAV2 ITR
     promoter        complement(1876..1894)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1904..1921)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1904..1920)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1913..1935)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(2061..2516)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2198..2219)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2410..2429
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2543..2647
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2648..3508
                     /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(2866..2885)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3679..4267
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4168..4187
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4421..4438
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4555..4576
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4591..4621
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4629..4645
                     /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     4634..4656
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4653..4669
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4653..4669
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4688..4708
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4690..4708
                     /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 catggcaaat aaaacgtaca agataggcaa aaacgctggg
     1081 tacgatgggt gtggtctgtg cctcgctgcg atcagcgaga atgaggctat caaggtcaaa
     1141 tacttgcgcg acatctgccc ggactacgat ggcgacgaca aagctgaaga ttggttgagg
     1201 tgggggacgg attctcgcgt taaggccgct gcgcttgaaa tggagcaata tgcctacaca
     1261 tccgtcggta tggcgtcctg ttgggagttc gttgagttgt gactcgagcg gccgccactg
     1321 tgctggatat ctgcagaatt caaacaccat tgtcacactc cattatatca aacaccattg
     1381 tcacactcca ttaattagag acgagctcgt ctctagctta agtttaaacc gctgatcagc
     1441 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     1501 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     1561 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     1621 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg gtcgacgata
     1681 tcagatctga ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     1741 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     1801 tgagcgagcg agcgcgcaga gagggagtgg ccsccmcccc cccccccccc tgcattctag
     1861 agagctccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac
     1921 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     1981 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     2041 gcagcctgaa tggcgaatgg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     2101 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     2161 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     2221 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     2281 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     2341 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     2401 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     2461 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     2521 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     2581 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     2641 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     2701 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     2761 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     2821 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     2881 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     2941 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     3001 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     3061 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     3121 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     3181 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     3241 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     3301 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     3361 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     3421 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     3481 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     3541 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     3601 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     3661 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3721 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3781 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3841 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3901 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3961 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4021 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4081 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4141 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4201 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4261 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4321 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     4381 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     4441 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     4501 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     4561 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     4621 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     4681 caagcgcgca attaaccctc actaaaggga acaaaagctg ggtaccacta gtatgcatcc
     4741 tgcaggacgc gtcctaccca ctccctctat gcgcgctcgc tcactcactc ggccctggag
     4801 acctttggtc tccagggccg agtgagtgag cgagcgcgca tagagggagt gggtacctcc
     4861 atcatctggt ttcccagatc tgatatcggc gcgcccgagt ccaacacccg tgggaatccc
     4921 atgggcacca tggcccctcg ctccaaaaat gctttcgcgt cgcgcagaca ctgctcggta
     4981 gtttcgggga tcagcgtttg agtaagagcc cgcgtctgaa ccctccgcgc cgccccggcc
     5041 ccagtggaaa gacgcgcagg caaaacgcac cacgtgacgg agcgtgaccg cgcgccgagc
     5101 gcgcgccaag gtcgggcagg aa
//
