LOCUS       62056_22545        8958 bp DNA     circular SYN 02-NOV-2019
DEFINITION  Expression vector pVG185_w2-y1, complete sequence.
ACCESSION   MN551090
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8958)
  AUTHORS   Lopez del Amo V Sr., Bishop AL Sr., Sanchez C HM, Bennett JB, Feng 
            X, Marshall JM, Bier E, Gantz VM.
  TITLE     Trans-complementing split-gene drive system provides flexible 
            application for safer laboratory investigation and potential field 
            deployment
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8958)
  AUTHORS   Lopez del Amo V Sr., Bishop AL Sr., Sanchez C HM, Bennett JB, Feng 
            X, Marshall JM, Bier E, Gantz VM.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-OCT-2019) Biological Sciences, University of 
            California San Diego, 9500 Gilman Dr, Bonner Hall 4305, La Jolla, CA
            92093, USA
REFERENCE   3  (bases 1 to 8958)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (08-OCT-2019) Biological Sciences, University of California San 
            Diego, 9500 Gilman Dr, Bonner Hall 4305, La Jolla, CA 92093, USA"
FEATURES             Location/Qualifiers
     source          1..8958
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    877..898
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        913..943
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1065..2092
                     /label=white homology arm
                     /note="white homology arm"
     promoter        2098..2894
                     /label=dU6-1 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-1
                     snRNA (Port et al., 2014)"
     misc_feature    2894..2913
                     /label=w2 gRNA
                     /note="w2 gRNA"
     misc_RNA        2914..2989
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2990..3410
                     /label=dU6-3 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-3
                     snRNA (Port et al., 2014)"
     misc_feature    3410..3429
                     /label=yellow gRNA
                     /note="yellow gRNA"
     misc_feature    3430..3505
                     /label=gRNA scaffold
                     /note="gRNA scaffold"
     misc_feature    3512..3813
                     /label=U6-3_3'UTR
                     /note="U6-3_3'UTR"
     polyA_signal    complement(3840..3961)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4087..4803)
                     /label=EGFP
                     /note="enhanced GFP"
     regulatory      4858..4982
                     /label=Hsp70 promoter
                     /note="Hsp70 promoter"
                     /regulatory_class="promoter"
     misc_feature    5014..5057
                     /label=3xP3 eye promoter
                     /note="3xP3 eye promoter"
     misc_feature    5080..6091
                     /label=white homology arm
                     /note="white homology arm"
     primer_bind     complement(6147..6163)
                     /note="M13-forward. common sequencing primer, one of
                     multiple similar variants"
     promoter        complement(6162..6180)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6187..6203)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6344..6772
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             7116..7907
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             7928..8785
                     /label=AmpR
                     /note="beta-lactamase"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg gtaccgagct
     1021 cggatccact agtaacggcc gccagtgtgc tggaattcgc ccttgctcaa agtccccaaa
     1081 cgaaagccaa agatgccagc gatttacact cgcaattgtc ttatgaggtg gcaacatggt
     1141 ggaggaacct tgtcactccc accacccatc gtcccatcgt cccatcgtcc atcgcccaac
     1201 gacctgtgac cccggccccg tcaaccgtcc accgcccagc gcccactttc aaggccatcc
     1261 accaagtgcc agccctaatt tgcacatttt aaataggcct cgccatctct ttttttggga
     1321 aacttggaaa actcggatct tgggcatttt tattaaacag atgggctcct tctttataaa
     1381 gccgaggggg tggggtgggg gttagtgccc ggaaacaggg acggcggagc agggattagc
     1441 caggctgggc tagatttatg cacagacgcc ttcattttta gggacgaacg ccgtgaaatt
     1501 gaaaagtttt cagcttgaaa ccttttctag atgcacaaaa aataaataaa agtataaacc
     1561 tacttcgtag gatacttcgt tttgttcggg gttagatgag cataacgctt gtagttgata
     1621 tttgagatcc cctatcattg cagggtgaca gcggagcggc ttcgcagagc tgcattaacc
     1681 agggcttcgg gcaggccaaa aactacggca cgctcctgcc acccagtccg ccggaggact
     1741 ccggttcagg gagcggccaa ctagccgaga acctcaccta tgcctggcac aatatggaca
     1801 tctttggggc ggtcaatcag ccgggctccg gatggcggca gctggtcaac cggacacgcg
     1861 gactattctg caacgagcga cacataccgg cgcccaggaa acatttgctc aagaacggtg
     1921 agtttctatt cgcagtcggc tgatctgtgt gaaatcttaa taaagggtcc aattaccaat
     1981 ttgaaactca gtttgcggcg tggcctatcc gggcgaactt ttggccgtga tgggcagttc
     2041 cggtgccgga aagacgaccc tgctgaatgc ccttgccttt cgatcgccgc agacgtcatt
     2101 ttcaacgtcc tcgatagtat agtggttagt atccccgcct gtcacgcggg agaccggggt
     2161 tcaattcccc gtcggggaga atctgtgatt cttttttttt ttcttttact ttgttatata
     2221 aacaattttt gttttaattg aatctaattt gccattgctt ttaggaatct caggcatcca
     2281 gcaagcgttt gtccgccgaa tcgcccatca gtgaagaaga tcctgtggcg gctacgaaaa
     2341 tctccccggc catgtcggcc tccacctcca gcgaaaaacc catcagcgag ctggccacct
     2401 ctgtgctgac ccaccgcttt ccagactcca cctcctcacc cggcgaacat ggccttggac
     2461 gaatgcagtt gtcgatccgc tacagcgccc agcgtcaaaa actagacgtg accatacaca
     2521 aaatccagaa gataccactt cgcgatccca gcaatatccc cgatccgtat gttaagctgt
     2581 atctgttgcc tggacgcacc aaggagtcga aacgcaagac gagcgtgatc aaggacaact
     2641 gcaaccccgt ctacgatgca tcctttgagt acctgatttc cattgccgaa ctcaggcaga
     2701 cggaactgga ggtgacggtg tgcacccaaa agggattcct atccggcggt agtcccatca
     2761 ttggcatggt aggtacccga aagcaacccc ttagttacag acacagcgcg tacgtccttc
     2821 gcatccttat gattcccaag tacatattct gcaagagtac agtatatata ggaaagatat
     2881 ccgggtgaac ttcggcgata cttggatgcc ctggttttag agctagaaat agcaagttaa
     2941 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttttgctca
     3001 cctgtgattg ctcctactca aatacaaaaa catcaaattt tctgtcaata aagcatattt
     3061 atttatattt attttacagg aaagaattcc ttttaaagtg tattttaacc tataatgaaa
     3121 aacgattaaa aaaaatacat aaaataattc gaaaattttt gaatagccca ggttgataaa
     3181 aattcatttc atacgtttta taacttatgc ccctaagtat tttttgacca tagtgtttca
     3241 attctacatt aattttacag agtagaatga aacgccacct actcagccaa gaggcgaaaa
     3301 ggttagctcg ccaagcagag agggcgccag tgctcactac tttttataat tctcaacttc
     3361 tttttccaga ctcagttcgt atatatagac ctattttcaa tttaacgtcg gttttggaca
     3421 ctggaaccgg ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact
     3481 tgaaaaagtg gcaccgagtc ggtgcttttt tgcctacctg gagcctgaga gttgttcaat
     3541 aaaataaaaa tgtttcgttt ttttgctttc gccagtattt attatttttc atcaatatgt
     3601 attcaatttg gtatgtattt agtaattgta atatatagac aatggttttc cgttgacgta
     3661 catacatctg acgtgtgttt atttagacat aatagttatg ttttcacatc tttttaatgt
     3721 tcgcttaatg cgtatgcatt ctagacaatt gtgctcggca acagtatatt tgtggtgtgc
     3781 caaccaacaa cctgcaggag ctccagcttt tgtgctagcg tacgcgtatc gataagcttt
     3841 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     3901 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     3961 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     4021 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatctag agtcgcggcc
     4081 gctttacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag
     4141 caggaccatg tgatcgcgct tctcgttggg gtctttgctc agggcggact gggtgctcag
     4201 gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg
     4261 gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt tcaccttgat
     4321 gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt tgtactccag
     4381 cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt
     4441 caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt
     4501 gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg
     4561 ctgcttcatg tggtcggggt agcggctgaa gcactgcacg ccgtaggtca gggtggtcac
     4621 gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca gggtcagctt
     4681 gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc
     4741 gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac
     4801 catggtggcg accggtggat cccgggcccg cggtaccgtc gactctagcg gtaccccgat
     4861 tgtttagctt gttcagctgc gcttgtttat ttgcttagct ttcgcttagc gacgtgttca
     4921 ctttgcttgt ttgaattgaa ttgtcgctcc gtagacgaag cgcctctatt tatactccgg
     4981 cggtcgaggg ttcgaaatcg ataagcttgg atcctaattg aattagctct aattgaatta
     5041 gtctctaatt gaattagatc cccgggcgag ctcgcatatg gcatccaagt atcgccatcc
     5101 gggatgcgac tgctcaatgg ccaacctgtg gacgccaagg agatgcaggc caggtgcgcc
     5161 tatgtccagc aggatgacct ctttatcggc tccctaacgg ccagggaaca cctgattttc
     5221 caggccatgg tgcggatgcc acgacatctg acctatcggc agcgagtggc ccgcgtggat
     5281 caggtgatcc aggagctttc gctcagcaaa tgtcagcaca cgatcatcgg tgtgcccggc
     5341 agggtgaaag gtctgtccgg cggagaaagg aagcgtctgg cattcgcctc cgaggcacta
     5401 accgatccgc cgcttctgat ctgcgatgag cccacctccg gactggactc atttaccgcc
     5461 cacagcgtcg tccaggtgct gaagaagctg tcgcagaagg gcaagaccgt catcctgacc
     5521 attcatcagc cgtcttccga gctgtttgag ctctttgaca agatccttct gatggccgag
     5581 ggcagggtag ctttcttggg cactcccagc gaagccgtcg acttcttttc ctagtgagtt
     5641 cgatgtgttt attaagggta tctagcatta cattacatct caactcctat ccagcgtggg
     5701 tgcccagtgt cctaccaact acaatccggc ggacttttac gtacaggtgt tggccgttgt
     5761 gcccggacgg gagatcgagt cccgtgatcg gatcgccaag atatgcgaca attttgccat
     5821 tagcaaagta gcccgggata tggagcagtt gttggccacc aaaaatttgg agaagccact
     5881 ggagcagccg gagaatgggt acacctacaa ggccacctgg ttcatgcagt tccgggcggt
     5941 cctgtggcga tcctggctgt cggtgctcaa ggaaccactc ctcgtaaaag tgcgacttat
     6001 tcagacaacg gtgagtggtt ccagtggaaa caaatgatat aacgcttaca attcttggaa
     6061 acaaattcgc tagattttag atagaattgc caagggcgaa ttctgcagat atccatcaca
     6121 ctggcggccg ctcgagcatg catctagagg gcccaattcg ccctatagtg agtcgtatta
     6181 caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     6241 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     6301 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc
     6361 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     6421 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     6481 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     6541 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     6601 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     6661 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     6721 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     6781 attcagggcg caagggctgc taaaggaagc ggaacacgta gaaagccagt ccgcagaaac
     6841 ggtgctgacc ccggatgaat gtcagctact gggctatctg gacaagggaa aacgcaagcg
     6901 caaagagaaa gcaggtagct tgcagtgggc ttacatggcg atagctagac tgggcggttt
     6961 tatggacagc aagcgaaccg gaattgccag ctggggcgcc ctctggtaag gttgggaagc
     7021 cctgcaaagt aaactggatg gctttcttgc cgccaaggat ctgatggcgc aggggatcaa
     7081 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     7141 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     7201 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     7261 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     7321 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     7381 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatcc caccttgctc
     7441 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     7501 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     7561 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     7621 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     7681 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     7741 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     7801 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     7861 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga attgaaaaag
     7921 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     7981 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     8041 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     8101 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     8161 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     8221 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     8281 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     8341 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     8401 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     8461 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     8521 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     8581 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     8641 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     8701 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     8761 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     8821 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     8881 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     8941 aaagatcaaa ggatcttc
//
