LOCUS       62056_22600        9522 bp DNA     circular SYN 26-APR-2021
DEFINITION  Cloning vector pVMG0109_CC-U6-3-w5_GFP_wHAs_O, complete sequence.
ACCESSION   MW925706
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9522)
  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  JOURNAL   Unpublished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 9522)
  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 9522)
  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            "Unpublished"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..9522
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..429
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             773..1564
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             1585..2442
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      2616..3204
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    3492..3513
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3528..3558
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3566..3582
                     /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     3590..3606
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3680..4724
                     /label=white homology arm (left)
                     /note="white homology arm (left)"
     3'UTR           4746..6414
                     /label=3'UTR of Dmel actin5c (FBtr0345894)
                     /note="3'UTR of Dmel actin5c (FBtr0345894)"
     5'UTR           6422..6821
                     /label=regulatory region of Dmel U6-3
                     /note="regulatory region of Dmel U6-3"
     misc_feature    6822..6841
                     /label=Dmel white gRNA-5
                     /note="Dmel white gRNA-5"
     misc_RNA        6842..6917
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     3'UTR           6918..7017
                     /label=3'UTR of Dmel U6-3
                     /note="3'UTR of Dmel U6-3"
     polyA_signal    complement(7044..7165)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(7292..7999)
                     /label=mNeonGreen
                     /note="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
     regulatory      complement(8000..8270)
                     /label=3xP3 promoter
                     /note="3xP3 promoter"
                     /regulatory_class="promoter"
     misc_feature    8276..9270
                     /label=white homology arm (right)
                     /note="white homology arm (right)"
     promoter        complement(9341..9359)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9366..9382)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
       61 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      181 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaatt cagggcgcaa gggctgctaa aggaagcgga acacgtagaa
      481 agccagtccg cagaaacggt gctgaccccg gatgaatgtc agctactggg ctatctggac
      541 aagggaaaac gcaagcgcaa agagaaagca ggtagcttgc agtgggctta catggcgata
      601 gctagactgg gcggttttat ggacagcaag cgaaccggaa ttgccagctg gggcgccctc
      661 tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttcttgccgc caaggatctg
      721 atggcgcagg ggatcaagat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
      781 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
      841 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
      901 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
      961 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     1021 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     1081 gtcatcccac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     1141 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     1201 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     1261 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     1321 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     1381 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     1441 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     1501 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     1561 cttctgaatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     1621 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     1681 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     1741 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     1801 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     1861 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     1921 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     1981 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     2041 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     2101 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     2161 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     2221 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     2281 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     2341 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     2401 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     2461 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     2521 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     2581 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     2641 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     2701 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     2761 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     2821 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     2881 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     2941 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     3001 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     3061 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     3121 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     3181 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     3241 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     3301 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     3361 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     3421 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     3481 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     3541 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     3601 tatgaccatg attacgccaa gcttggtacc gagctcggat ccactagtaa cggccgccag
     3661 tgtgctggaa ttcgcccttg ctcaaagtcc ccaaacgaaa gccaaagatg ccagcgattt
     3721 acactcgcaa ttgtcttatg aggtggcaac atggtggagg aaccttgtca ctcccaccac
     3781 ccatcgtccc atcgtcccat cgtccatcgc ccaacgacct gtgaccccgg ccccgtcaac
     3841 cgtccaccgc ccagcgccca ctttcaaggc catccaccaa gtgccagccc taatttgcac
     3901 attttaaata ggcctcgcca tctctttttt tgggaaactt ggaaaactcg gatcttgggc
     3961 atttttatta aacagatggg ctccttcttt ataaagccga gggggtgggg tgggggttag
     4021 tgcccggaaa cagggacggc ggagcaggga ttagccaggc tgggctagat ttatgcacag
     4081 acgccttcat ctttagggac gaacgccgtg aaattgaaaa gttttcagct tgaaaccttt
     4141 tctagatgca caaaaaataa ataaaagtat aaacctactt cgtaggatac ttcgttttgt
     4201 tcggggttag atgagcataa cgcttgtagt tgatatttga gatcccctat cattgcaggg
     4261 tgacagcgga gcggcttcgc agagctgcat taaccagggc ttcgggcagg ccaaaaacta
     4321 cggcacgctc ctgccaccca gtccgccgga ggactccggt tcagggagcg gccaactagc
     4381 cgagaacctc acctatgcct ggcacaatat ggacatcttt ggggcggtca atcagccggg
     4441 ctccggatgg cggcagctgg tcaaccggac acgcggacta ttctgcaacg agcgacacat
     4501 accggcgccc aggaaacatt tgctcaagaa cggtgagttt ctattcgcag tcggctgatc
     4561 tgtgtgaaat cttaataaag ggtccaatta ccaatttgaa actcagtttg cggcgtggcc
     4621 tatccgggcg aacttttggc cgtgatgggc agttccggtg ccggaaagac gaccctgctg
     4681 aatgcccttg cctttcgatc gccgcagggc atccaagtat cgccacgtcg taaggctagt
     4741 tttgagaagg atcgcttgtc tgggcaagag gatcaggatc gggatggtct tgattctgct
     4801 ggaggaggag gaggagaagt cgaggaagca gcagcgaaag tgcaagtgcg agtggtggaa
     4861 gtttggagtg cagcacaaca aaatcaacaa caacaccaac tacaagatga aaagagcgga
     4921 accacctgca caccatcatc attatcatca tcgttttggg cgcatgttgt gtggttccag
     4981 cgtattaata taattaattt attccacatg agatatgata tgatatacta tgtatttttt
     5041 gttttttttt tatttgtaaa cctttaatat aacaagaact acaaaaaatg aaaatgagcg
     5101 aaaatgcata ttctgccatt ccacacacac accaacaaca cccaacacac gcacacccac
     5161 aagcttacac acacacacat tcgcggcatg acaaggacat caagataaag aagaacttaa
     5221 agaagatatt tcccaaagcg caaaaagaac acacacacat tgcaaaacac aaacaacaca
     5281 ctagcgtttt gtacaattcg tcagcaacct tatgtattat tttttaatta tgatgtaatt
     5341 ataaacaaag tgaaacaaaa atatgaaaac aaaaaggaaa atcaaatctg tcttctcttt
     5401 ctcccgctct cctctcgctc tctgctgcta acctcgccct ctcctctctc acctttttgt
     5461 ctgtctctct tccacttttt tgccggccgg caaaataacc cacacacact cacacttggc
     5521 tgcagtttcg cgtgcgatat tcacacacat tcaagcatac aagcatacat acatatgtat
     5581 ttttttttta tttgtacact tttctaattg catgcgtatc gattgataag tttacgcctg
     5641 aaaatgttaa ttaaaatgtg aaaatgcaac tgaaaaactg atgaaatgaa acaacaacaa
     5701 gcgaacaatt tgctacatgt gtattgtcta acaaccgtta ctgcaacggt tgcttcggaa
     5761 aagggtgaag aggaagaggg cacgaggtcg acgaactaag gctgcgacga caagcggctg
     5821 tctcgctcgc acgcacgcac gcatgcgggc taatgatgcg attaaagtgc cgttttaaat
     5881 ggccaagatc caaataaaag tcgagataat aaggcaaaca cgctaacaca aatcgctggc
     5941 atagtttcat ccgccagttt ggccaaaagc agcggctgca agcgccgttt atccggtttt
     6001 tcccaaacat tttccattca tgcggccaaa gaggaggagc acccatcttt ccatctttct
     6061 ctgactcgca cgagcaatgc agtgcaccgg aaagaaagtg gtgcgaattg ggatttatag
     6121 acttacttta tagacttaca cgtacataca tacatagata ttatattatt taaatacatt
     6181 tttattatat tgtttatata ttagttcgac ttcgatatca tttctgccag tataattatg
     6241 ggctctgctt gatttggcgc tcggatgcta cggtaaaagt tatctttctg tgctttctcc
     6301 tttgattctt gagtttcaac ggttccgttt ttgttgatct cgttcgtctt cttcttcttg
     6361 cgttctagac gttctccttt gttttttgtt cacaacggat ttcgtagcac cttggctagc
     6421 ggctcctact caaatacaaa aacatcaaat tttctgtcaa taaagcatat ttatttatat
     6481 ttattttaca ggaaagaatt ccttttaaag tgtattttaa cctataatga aaaacgatta
     6541 aaaaaaatac ataaaataat tcgaaaattt ttgaatagcc caggttgata aaaattcatt
     6601 tcatacgttt tataacttat gcccctaagt attttttgac catagtgttt caattctaca
     6661 ttaattttac agagtagaat gaaacgccac ctactcagcc aagaggcgaa aaggttagct
     6721 cgccaagcag agagggcgcc agtgctcact actttttata attctcaact tctttttcca
     6781 gactcagttc gtatatatag acctattttc aatttaacgt cggcatccaa gtatcgccat
     6841 cgttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag
     6901 tggcaccgag tcggtgcttt ttttgcctac ctggagcctg agagttgttc aataaaataa
     6961 aaatgtttcg tttttttgct ttcgccagta tttattattt ttcatcaata tgtattcgct
     7021 agcgtacgcg tatcgataag ctttaagata cattgatgag tttggacaaa ccacaactag
     7081 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     7141 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     7201 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc
     7261 tgattatgat ctagagtcgc ggcccctatt acttatacag ttcatccatt cccatcacgt
     7321 cggtgaacgc cttttgccat tctttgaaat tcaattctgt tttggaatgt ttcagctccg
     7381 tcttgcgaaa aacatacata ggttgatttt tcagataatt cgcagccatg ggtttcgcaa
     7441 acgtgtatgt agtgcgcgcc gtggaacggt agcgcttgcc atttcccgtt gtgtagctcc
     7501 acttgaatgt gctaatgatg gtcttatcat ttgggtaggt cttcttgcta cggcaccagt
     7561 ctgcagcggt caagctgttc gtcataactg ggccgtccgc ggggaatcca gtacccttta
     7621 cctgagcttc tcctttgatg tggctaccct catatgtata gcggtagttc accgtcagcg
     7681 aggctccatc ctcgaactgc attgtgcggt gcacctggta gccggagccg tccaccatcg
     7741 ccgcttgaaa gggcgacatg ccgtccggat atggcagata ctgatgaaat ccatatccaa
     7801 tatggggaac caagatccag gggctgaatt gcaggtcgcc ttttgtgctt ttcagattca
     7861 attcttcata gccgtcattg gggttgcctg taccttgtcc caccatatca aaatccactc
     7921 cattaatgga gccaaagata tgcaactcat gtgtagccgg cagggatgcc atgttgtcct
     7981 cctcgccctt cgacaccatg gtggcgaccg gtggatcccg ggcccgcggt accgtcgact
     8041 ctagcggtac cccgattgtt tagcttgttc agctgcgctt gtttatttgc ttagctttcg
     8101 cttagcgacg tgttcacttt gcttgtttga attgaattgt cgctccgtag acgaagcgcc
     8161 tctatttata ctccggcggt cgagggttcg aaatcgataa gcttggatcc taattgaatt
     8221 agctctaatt gaattagtct ctaattgaat tagatccccg ggcgagctcg catatatccg
     8281 ggatgcgact gctcaatggc caacctgtgg acgccaagga gatgcaggcc aggtgcgcct
     8341 atgtccagca ggatgacctc tttatcggct ccctaacggc cagggagcac ctgattttcc
     8401 aggccatggt gcggatgcca cgacatctga cctatcggca gcgagtggcc cgcgtggatc
     8461 aggtgatcca ggagctttcg ctcagcaaat gtcagcacac gatcatcggt gtgcccggca
     8521 gggtgaaagg tctgtccggc ggagaaagga agcgtctggc attcgcctcc gaggcactaa
     8581 ccgatccgcc gcttctgatc tgcgatgagc ccacctccgg actggactca tttaccgccc
     8641 acagcgtcgt ccaggtgctg aagaagctgt cgcagaaggg caagaccgtc atcctgacca
     8701 ttcatcagcc gtcttccgag ctgtttgagc tctttgacaa gatccttctg atggccgagg
     8761 gcagggtagc tttcttgggc actcccagcg aagccgtcga cttcttttcc tagtgagttc
     8821 gatgtgttta ttaagggtat ctagcattac attacatctc aactcctatc cagcgtgggt
     8881 gcccagtgtc ctaccaacta caatccggcg gacttttacg tacaggtgtt ggccgttgtg
     8941 cccggacggg agatcgagtc ccgtgatcgg atcgccaaga tatgcgacaa ttttgccatt
     9001 agcaaagtag cccgggatat ggagcagttg ttggccacca aaaatttgga gaagccactg
     9061 gagcagccgg agaacgggta cacctacaag gccacctggt tcatgcagtt ccgggcggtc
     9121 ctgtggcgat cctggctgtc ggtgctcaag gaaccactcc tcgtaaaagt gcgacttatt
     9181 cagacaacgg tgagtggttc cagtggaaac aaatgatata acgcttacaa ttcttggaaa
     9241 caaattcgct agattttaga tagaattgcc aagggcgaat tctgcagata tccatcacac
     9301 tggcggccgc tcgagcatgc atctagaggg cccaattcgc cctatagtga gtcgtattac
     9361 aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     9421 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     9481 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gg
//
