LOCUS       62056_22570        4861 bp DNA     circular SYN 07-OCT-2020
DEFINITION  Cloning vector pVG362_Aste-U6a-Swap3-gRNA, complete sequence.
ACCESSION   MW030449
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4861)
  AUTHORS   Adolfi A, Gantz V, Jasinskiene N, Lee H-F., Hwang K, Terradas G, 
            Bulger E, Ramaiah A, Bennett J, Emerson JJ, Marshall J, Bier E.
  TITLE     Efficient population modification gene-drive rescue system in the 
            malaria mosquito Anopheles stephensi
  JOURNAL   Nat Commun (2020) In press
REFERENCE   2  (bases 1 to 4861)
  AUTHORS   Adolfi A, Gantz V, Jasinskiene N, Lee H-F., Hwang K, Terradas G, 
            Bulger E, Ramaiah A, Bennett J, Emerson JJ, Marshall J, Bier E.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-SEP-2020) Developmental and Stem Cell Biology, 
            University of California, San Francisco, 1650 Owens St., San 
            Francisco, CA 94158, USA
REFERENCE   3  (bases 1 to 4861)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Commun 
            (2020) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (22-SEP-2020) Developmental and Stem Cell Biology, University of 
            California, San Francisco, 1650 Owens St., San Francisco, CA 94158, 
            USA"
FEATURES             Location/Qualifiers
     source          1..4861
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(944..1801)
                     /label=AmpR
                     /note="beta-lactamase"
     CDS             complement(1822..2613)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(2957..3385)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     3526..3542
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3549..3567
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    4065..4084
                     /label=Swap3 Target
                     /note="Swap3 Target"
     misc_RNA        4085..4160
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(4641..4657)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4665..4681)
                     /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(4689..4719)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4734..4755)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaattc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg
     1861 cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag
     1921 ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag
     1981 ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca
     2041 ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc
     2101 gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag
     2161 accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg
     2221 gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt
     2281 ctcggcagga gcaaggtggg atgacaggag atcctgcccc ggcacttcgc ccaatagcag
     2341 ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt
     2401 ggccagccac gatagccgcg ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc
     2461 ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga
     2521 gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg
     2581 agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg
     2641 atcagatctt gatcccctgc gccatcagat ccttggcggc aagaaagcca tccagtttac
     2701 tttgcagggc ttcccaacct taccagaggg cgccccagct ggcaattccg gttcgcttgc
     2761 tgtccataaa accgcccagt ctagctatcg ccatgtaagc ccactgcaag ctacctgctt
     2821 tctctttgcg cttgcgtttt cccttgtcca gatagcccag tagctgacat tcatccgggg
     2881 tcagcaccgt ttctgcggac tggctttcta cgtgttccgc ttcctttagc agcccttgcg
     2941 ccctgaattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     3001 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     3061 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     3121 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     3181 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     3241 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     3301 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     3361 cttaatgcgc cgctacaggg cgcgtccatt cgccattcag gctgcgcaac tgttgggaag
     3421 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     3481 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     3541 gtgaattgta atacgactca ctatagggcg aattgggccc tctagatgca tgctcgagcg
     3601 gccgccagtg tgatggatat ctgcagaatt cgcccttaag cttatcgata cgcgtacctc
     3661 aactgccgac gagttgctcg ggtttcgatg agttccagca tacgcgcatg ctgctgagtg
     3721 cttcttggtc gcatatacac tcatgccctt tttgccttgt gacgattttt ttttttctac
     3781 tcaatgcaaa tttcaaattt gagggcaatt ttcgtgctta tgtttaataa tctataaaag
     3841 aaaatttaaa actaaaaact catgtacagc agtagatgaa gctttcaccg ttcttctcac
     3901 ttaaattcta cctgcacatt ggctggtgga agagctataa ttgaagcaga accttataac
     3961 agtcgcgcgg aaggatcaag agctttgtga atatccttta tccatcgcta gaactaaaac
     4021 ggatgacaga cgggatatat accaacccat ttgccccctc ccttgttctt ggaggagcgc
     4081 accagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa
     4141 aagtggcacc gagtcggtgc tttttttgtg gaaatttgat tcacttgttt taggaaaata
     4201 attaccttcc tttgaaacag gtatttacaa tgatggacga tagaaaaaga ccaccttaaa
     4261 atgctttatt caagctattg ctttcgattc ttcattgaga tctaatctca aatttgtgta
     4321 tttagaaaaa acagcgaata ttgtactata ataattcttt ggtaatctca ggctttcagt
     4381 aatgatagaa tccttaagac tgtaagaata tacaagtttt aaaaaaagga aaacaggtat
     4441 atcaaatcag aaaacaacaa cagctctttt ccgttgaatt ctacctgtac gatggcttaa
     4501 ggatgagctg taactgaaac gaaaccttac aacagccacg cagaacgtag gccggccgat
     4561 ctcggataag ggcgaattcc agcacactgg cggccgttac tagtggatcc gagctcggta
     4621 ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     4681 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4741 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4801 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4861 t
//
