LOCUS       62056_141        7475 bp DNA     circular SYN 14-JUN-2021
DEFINITION  Cloning vector 3xP3_attB_eGFP_Vasa-AcrIIA4, complete sequence.
ACCESSION   MZ172909
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7475)
  AUTHORS   Taxiarchi C, Beaghton A, Illansinhage Don N, Kyrou K, Gribble M, 
            Shittu D, Collins SP, Beisel CL, Galizi R, Crisanti A.
  TITLE     A genetically encoded anti-CRISPR protein constrains gene drive 
            spread and prevents population suppression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7475)
  AUTHORS   Taxiarchi C, Beaghton A, Illansinhage Don N, Kyrou K, Gribble M, 
            Shittu D, Collins SP, Beisel CL, Galizi R, Crisanti A.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-MAY-2021) Life Sciences, Imperial College London, 
            Exhibition Road, London SW7 2AZ, UK
REFERENCE   3  (bases 1 to 7475)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (10-MAY-2021) Life Sciences, Imperial College London, Exhibition 
            Road, London SW7 2AZ, UK"
FEATURES             Location/Qualifiers
     source          1..7475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      869..1129
                     /label=3xp3
                     /note="3xp3"
                     /regulatory_class="promoter"
     protein_bind    1135..1168
                     /label=attB
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et
                     al., 2000)"
     CDS             1190..1906
                     /label=EGFP
                     /note="enhanced GFP"
     polyA_signal    2032..2153
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        2540..2644
                     /label=AmpR promoter
     CDS             2645..3502
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      3676..4264
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4552..4573
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4588..4618
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4626..4642
                     /label=lac operator
                     /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     4650..4666
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      4724..7014
                     /label=vasa2
                     /note="vasa2"
                     /regulatory_class="promoter"
     CDS             7024..7044
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
ORIGIN
        1 cttgcttttg tgacatgcat gattattctt tcatttactg acgtaacgat gtaaaacaca
       61 cagaagaagc atgacacaca gcaaagaatt gttcgttaat aaaacgttga tgaaaacttt
      121 gaaaacataa gaacttgcac ttttattcta taattcgtga aagcttgcac cgattgtctt
      181 tcattattca atgtaatgta ctgaaaggtg atttttcgca cttgtatact ctagaactga
      241 agtattctaa caatacgtca cctttaggtc cattccagga taaaatacac aagtgaaggt
      301 agttgtacaa agtacttaga ctaacccagg tttcaaaaaa gataacgacg aattgagcgt
      361 aactgcacaa agcccgttca ttgtcaacat atcgttcagg ggtcttcaaa ctttattatc
      421 agcccgcggg ccgcaacaac gttgcttaga gcaccatctt gatgtatcct ttatctagtg
      481 gcttatcgtt tatgtctcgt ttggctaaga aaatactaat aacatattca atttacggag
      541 cggtaccatg gtacaaacgt catactgcta gacttgacaa catgcccatc gagggttctt
      601 acctcaaatg gaccgatccc ttgtaactgg cggcgtaaac tcagtaaatt ccaaaagcca
      661 gtataggcca ccatgaccgt ctaactattc caaaaaggag aatatttgaa atttctatat
      721 ctatagtatt attactatat ctatatgttt ctttagtaca atattagagc ttgattattt
      781 tcctgaaaat tgtaaaatat gcatgttaga tggttgatac atcgttttaa aaaaaactgc
      841 tgttaatggc cacattttcg ggcgcgcccg cacggttccc acaatggtta attcgagctc
      901 gcccggggat ctaattcaat tagagactaa ttcaattaga gctaattcaa ttaggatcca
      961 agcttatcga tttcgaaccc tcgaccgccg gagtataaat agaggcgctt cgtctacgga
     1021 gcgacaattc aattcaaaca agcaaagtga acacgtcgct aagcgaaagc taagcaaata
     1081 aacaagcgca gctgaacaag ctaaacaatc ggggtaccgc tagagtcgat gcgggtgcca
     1141 gggcgtgccc ttgggctccc cgggcgcgta ctccacctca cccatcaaca tggtgagcaa
     1201 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     1261 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     1321 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1381 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     1441 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     1501 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     1561 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     1621 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     1681 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     1741 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     1801 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1861 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgcga
     1921 ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     1981 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     2041 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2101 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaagctct
     2161 gcagggatcc acgcgcctac tagtcggccg tacgggccct ttcgtctcgc gcgtttcggt
     2221 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     2281 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     2341 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     2401 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcggcctta agggcctcgt
     2461 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     2521 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     2581 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2641 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     2701 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     2761 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     2821 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     2881 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     2941 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     3001 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     3061 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     3121 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     3181 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     3241 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     3301 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     3361 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     3421 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     3481 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     3541 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     3601 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3661 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3721 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3781 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3841 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3901 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3961 atagttaccg gataaggcgc atcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4021 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4081 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4141 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4201 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4261 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4321 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     4381 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     4441 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     4501 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     4561 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     4621 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     4681 gcttgcatgc ctgcaggtcg actctagagg atccccgacc ggtatgtaga acgcgagcaa
     4741 attcttttcc ttccatgaca gcagcagcta cagtgggaag ccgaacgtca gacgtgtttg
     4801 acatgccgaa ctgggcggga aaattacagc gtgcgctttg ttttcaagca aatcacaact
     4861 cgctgcaaac aaaaccgttg agaaattgat tgttttataa tttgtattgt attttatttg
     4921 ttataataaa ctaaaaagac atactttttg catattttat acataaaaac atacatgcag
     4981 cattataaaa cacatataaa ccctccctgt agagtcccgt atcgaaatct tccatcctag
     5041 ttgcacagta cgacggacga gtaggccgtg tccgtgcaaa ttccagcttt tagcagtctt
     5101 ttgctcggag cactcgcggc gagtcggagg tttctgctga ggtgcttagc gctaaattag
     5161 ccaattgctt ttgcaagtga aataaccagc cgaatagtac ttcaaaactc aggtaagtga
     5221 actagtttta tagaacaaat gtttgtttgt tagaagttag tgaagtgttt gtgaaaaaaa
     5281 tctctcattt cggcaaaact aacgtaactg atttcaaatt gaattattgt tttgtgatgt
     5341 tatattattt catccagttg attagtattt tcttagttat gttcaaaata cagttaaatt
     5401 aaatttcatt tcatttactc ataaaataat ctcttggctt atttaatttt tctcgaattc
     5461 gcttgtattg ttcagtagca cgcgccattc gccctttgtt tcattttgta cctgctccca
     5521 ctaacacact ggcagtgcga aacaaaagcc ttcgcacgcg ttgctggtat tagagtgtgt
     5581 gcgtgtgtgt gttgagcgct ctgtcaaaat cggctgttgc cgccggtacc gaaattgcct
     5641 gttcgcacgc tgttcgtaaa cattccgtgg tgtgtatcgt gtgttgtgca tgttgcgcgc
     5701 ctcccccctt ttgatagcag gctgccgtgg ctgccgtggt gtgtggcgca gttgagtttt
     5761 tggattaatt ttctaaggaa atggcacgag aagagcggtg gcagtgtgtt ggtttgctct
     5821 gtcccttcct ttctgtgtga agtgttctta cagcacagca cgtatccacc accgcacaca
     5881 gagcaggcaa ggaagtggaa gtgaacaagt gtgctgcgca tgcatgtgtg tggggggcat
     5941 tttagctgag atcgtcgtta tttgagaagc ggtatagggg ccagtcggtg tcgacgtacg
     6001 gaagcggttt agttttaatc caagcgtatc ccgtcgtgga gtggttgtgt ggctctgtgt
     6061 gctctcatat cagttccaga gtgaggttag tagaatcaca gtccttggcc tttttcgtta
     6121 caagatatcc agaaggatgg cgttatttcc acagcttacc atggtgctct tgtttgctcg
     6181 aatcagggga gaaaaacagt ttcgtgtttc atgaaccgca gttggcactg gagcggattc
     6241 aaaagtcttc gatatgcaat agataagaga gtcgttgggg catagttggg aagcctttcc
     6301 gagatgtgga gtttccgaga ggagaaatgg tgctttcgtg cacgttccgg gacagcgggc
     6361 cccgcgaaga gcatctcgtt gtcgttcatc cggcaataat tgatgcgaaa agcgcgcgcg
     6421 ccactggctt agcgcagtgt acacagtgat attcacctac acacacagag gcacacgcct
     6481 tcacacgcgc gcgtgcttca aaggctactt cggtggcggt gtgtgaggtc gcttgcaatg
     6541 gacaatgaaa atttcgctgg aaaataccat cgtctcttta ggttgcaatg ggtgcgggta
     6601 gagcggtggt cgtcgatatt ggtggtgtag tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg
     6661 tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg
     6721 tgtgtgcaac ggcaattatt ttttgtaata tttcgaccat ctttctttct ctctctccac
     6781 gtgctgctgc tgttgctgct gctgctgcat tgcatgttcc actattcctc tcggtttgtg
     6841 cctgcggacg ccattgctag tcgaaagaga gtcgccgtta gtcgcgcttc gagcaacgga
     6901 cacgtttttt ggttgaaacc aacagctttt ttcatcttcg ggagacacac agatctcgaa
     6961 tcgtacattc ccataaggag aattgtcatc ttccggtgaa taaagaaagg aaacctcgag
     7021 atgccgaaga aaaagaggaa ggtgagcggc ggtagcaaca ttaatgatct catacgggag
     7081 atcaaaaaca aggattacac cgttaagctg agcggtacag actcgaacag tatcacgcag
     7141 ctgataatcc gcgtgaacaa cgacggaaac gaatacgtga tctcggagtc cgagaacgaa
     7201 agcattgtcg agaagtttat ctcggccttc aaaaatggct ggaatcaaga gtacgaagat
     7261 gaagaggaat tctataacga catgcagacg atcaccctga agtccgagtt gaattgatta
     7321 attaagcggc cgccttgggg tggggttgtt atgtgttgcg aacgagagtg gatctctctc
     7381 gacatttcct tatttttttt ttctattgtt aacacttaca acgaaacttc ggaagagaag
     7441 tttcctcatt cgaaacgagg agtcaaactc ctttc
//
