LOCUS       62056_22615        4423 bp DNA     circular SYN 26-APR-2021
DEFINITION  Cloning vector pVMG0164_Cq-U6-7_2xBbsI-gRNA, complete sequence.
ACCESSION   MW925704
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4423)
  AUTHORS   Feng X, Lopez Del Amo V, Mameli E, Lee M, Bishop AL, Perrimon N, 
            Gantz VM.
  TITLE     Optimized CRISPR tools and site-directed transgenesis towards gene 
            drive development in Culex quinquefasciatus mosquitoes
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4423)
  AUTHORS   Feng X, Lopez Del Amo V, Mameli E, Lee M, Bishop AL, Perrimon N, 
            Gantz VM.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-APR-2021) Biology Sciences, University of California, 
            San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA
REFERENCE   3  (bases 1 to 4423)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (05-APR-2021) Biology Sciences, University of California, San Diego,
            9500 Gilman Dr., La Jolla, CA 92093, USA"
FEATURES             Location/Qualifiers
     source          1..4423
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(16..604)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(778..1635)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(1636..1740)
                     /label=AmpR promoter
     primer_bind     2214..2230
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     5'UTR           2325..3339
                     /note="regulatory region of Culex quinquefasciatus U6-7
                     gene (CPIJ040693)"
     misc_feature    3340..3357
                     /note="2xBbsI spacer; gRNA insertion site"
     misc_RNA        3358..3433
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     3'UTR           3434..3978
                     /note="3'UTR of Culex quinquefasciatus U6-7 gene
                     (CPIJ040693)"
     primer_bind     complement(4037..4053)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4061..4077)
                     /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)."
     promoter        complement(4085..4115)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4130..4151)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
ORIGIN
        1 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
       61 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      121 aagctccctc gtgcgctctc ctgttccgac cctgtcgctt accggatacc tgtccgcctt
      181 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      241 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      301 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      361 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      421 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
      481 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      541 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      601 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
      661 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
      721 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
      781 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
      841 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
      901 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
      961 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1021 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     1081 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     1141 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     1201 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     1261 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1321 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     1381 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     1441 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1501 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     1561 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     1621 atgttgaata ctcatactct acctttttca atattattga agcatttatc agggttattg
     1681 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     1741 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     1801 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     1861 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     1921 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     1981 ctatgcggca tcagagcaga ttgtactgag agtgcaccaa atgcggtgtg aaataccgca
     2041 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     2101 ttgggaaggg cgatcggtgc gggcctcatc gctattacgc cagctggcga aagggggatg
     2161 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     2221 gacggccagt gcaacgcgat gacgatggat agcgattcat cgatgagctg acccgatcgc
     2281 cgccgccgga gggttgcgtt tgagacgggc gacagatggt ctcattcttc tcattgaaac
     2341 tgaattctgt taaaagggtt gtgtagggga catcttaaga tgacttcgct gaaaaaatct
     2401 cgttcctaga acaaatcctg ttattttggc agctgtctaa agttgaggta cgtttgtggc
     2461 caaaaatgac ctctcgaaaa atcatttttc taatacgttt gttagaattg ctcgaaaact
     2521 acccaaatgt ttgaaaatct ccacttcaaa cattgtagca tgtcaatacg attccctcga
     2581 acaagaaaca ccgttggatg acttgttttt accatatcgt tgtatttgag catcatttta
     2641 gtttcatttg acctaatgtc accccctcta aggggtgaga ttgggtcaat tttcaaacta
     2701 ttggcattca aggtagtgtt catcaaaatc caccatattt tgggaaaatg tcagtaaact
     2761 atctaagaac aaatctacgt tgaaagattt tcaatgttgt ttaaattgtt tttgttatta
     2821 agaaattacg agagttgtat tgttttttga cccatagtca cccccactga cggtactcgt
     2881 gttttggttt gttttataat gacctacacc ttgatagggc gggcggttcc ataaatgccc
     2941 attttcgtca attttcgcaa aaacctcagt ttttcaaaaa tcataacacc gcaccatttt
     3001 tgccgatttt agttgtctta gatgcaaatg aaaagtggtt aggtagcctt tcaaggaaat
     3061 ataatttgag gaccgataaa actagtcaaa catttgaaaa agtcgtatga aaacttaaaa
     3121 tggcgttttg accgtgagga ttccgagtta tgattttttt tcaaataaaa actcaaactt
     3181 tcggcgcgcc gcgcgccgaa ttgtgaaagt gacaaaaacg gggaaacaac ccatacaaca
     3241 acttttgtca ctaaaactgc gataacttta aaatttcagc aacattttat ttttaattga
     3301 ccaccaaggt cttaaggaag tgaataatag taaaatagcg ggtcttcgag aagacctgtt
     3361 ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc
     3421 accgagtcgg tgcaaaaaaa aaaaaaaagt tttcgagtga taaatagctc aaaaagaagt
     3481 gagcaaccaa gtttctatca aattttttgc aaaatttgtt gtttaaatag tgaaaatcaa
     3541 caaatataat ggagttagaa gcgagtgctt aacctgccaa acatacgaga attcccctta
     3601 ttaggttttt gttaaggggt tacatacatg tagaaaatca caaaatttaa tattacagaa
     3661 aatttgttca atcacttctg aaagattcat gaagatattt aatgattaaa ctgagtaaaa
     3721 gacgatttaa gttcacaatt ttgccatgcg caaaccgaac tgtcaaattt tgtgagcgta
     3781 tttctctgaa caccgagtta atttacgggt gccacgatat ctcgagatgg ataggaccaa
     3841 actggctgaa atttgaaggg aagattctca agacatatcc cgtgtgcatg acaaagatcg
     3901 atttttaaaa tttgggtttt ttgggttttt ttaatataaa aatcaaaaac ataagagagc
     3961 aactctctac caaaaccatg agaccattat cagttctgga ccagcgagct gtgctgcgac
     4021 tcgtggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     4081 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4141 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4201 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4261 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4321 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4381 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggc
//
