LOCUS       62056_22555        8976 bp DNA     circular SYN 02-NOV-2019
DEFINITION  Expression vector pVG304_e1-y1, complete sequence.
ACCESSION   MN551094
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8976)
  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  JOURNAL   Unpublished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 8976)
  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 8976)
  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            "Unpublished"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..8976
                     /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"
     promoter        1009..1027
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     misc_feature    1107..2116
                     /label=ebony homology arm
                     /note="ebony homology arm"
     promoter        2123..2919
                     /label=dU6-1 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-1
                     snRNA (Port et al., 2014)"
     misc_feature    2919..2938
                     /label=e1 gRNA
                     /note="e1 gRNA"
     misc_RNA        2939..3014
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3015..3435
                     /label=dU6-3 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-3
                     snRNA (Port et al., 2014)"
     misc_feature    3435..3454
                     /label=y1 gRNA
                     /note="y1 gRNA"
     misc_feature    3455..3530
                     /label=gRNA scaffold
                     /note="gRNA scaffold"
     misc_feature    3537..3838
                     /label=U6-3_3'UTR
                     /note="U6-3_3'UTR"
     polyA_signal    complement(3865..3986)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4112..4828)
                     /label=EGFP
                     /note="enhanced GFP"
     regulatory      4883..5007
                     /label=Hsp70 promoter
                     /note="Hsp70 promoter"
                     /regulatory_class="promoter"
     misc_feature    5039..5082
                     /label=3xP3 eye promoter
                     /note="3xP3 eye promoter"
     misc_feature    5106..6109
                     /label=ebony homology arm
                     /note="ebony homology arm"
     primer_bind     complement(6147..6163)
                     /note="M13-forward. common sequencing primer, one of
                     multiple similar variants"
     promoter        complement(6180..6198)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6205..6221)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6362..6790
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             7134..7925
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             7946..8803
                     /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 gccaagctat ttaggtgaca
     1021 ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcca ctagtaacgg
     1081 ccgccagtgt gctggaattc gcccttagtt agcatgccgc attccgagcg cttgtttagg
     1141 cttttggcca agtccaatgt ccgagataca acagttgagc cagcgtaact ccttcaagat
     1201 gaggcatttt atggagctag gttattcaaa actcgaaaat tattattttt atttttaagg
     1261 cacatttttt taggaataaa tacctggcaa tgtaaacgtg gattttcaaa gccggaacac
     1321 atctcaattc atagtgtaag cccacataga ttaaactgac cgtaattagt aacgacaagc
     1381 aggtggctat gctaatgcat taagtaatat ttactaaagt taaggagctc tctagtctcc
     1441 tattagacaa acctgtaatg aaagtatgac tgataggtta gggaaaaata tggtgctaag
     1501 cgaatagata aaaatgtcgc cataatttaa aataacatgt gcctgttgct aagcgtttag
     1561 tcgcaaagaa gaaaaactaa cttaactatt actgaaaata aaatgaatga gtctttatac
     1621 tccacgccat ttagcgagaa ataagaattc caactatttc cataaggcta ttcttaagta
     1681 ctgatataaa taataaaaca ttaataaaat cgtattaccc tgaaaccctt aattatcaag
     1741 tcattaacat ggaatatcga attccagttt acttgaactt gcaatctctg cagcttaaga
     1801 gacaacaatt taattgaata aatatagctc aacagaagcc atccgtttaa gccagctccc
     1861 atattcatac tcattcatta taatagttta ttgtgcgagc gctccattaa cacgaagtgt
     1921 tataaatatg catgtatgaa tgaatttaat tgctaccgca aggcagcaag gcattagcct
     1981 gcattgcata atacgagctg ggaatgatga tgtaacctag ggattctccg actgagattc
     2041 taagcccaaa actaacaaag tattccccac agttaatata tcttcaagat gggttcgctg
     2101 ccacaattgt cgatcgaccg gtattttcaa cgtcctcgat agtatagtgg ttagtatccc
     2161 cgcctgtcac gcgggagacc ggggttcaat tccccgtcgg ggagaatctg tgattctttt
     2221 tttttttctt ttactttgtt atataaacaa tttttgtttt aattgaatct aatttgccat
     2281 tgcttttagg aatctcaggc atccagcaag cgtttgtccg ccgaatcgcc catcagtgaa
     2341 gaagatcctg tggcggctac gaaaatctcc ccggccatgt cggcctccac ctccagcgaa
     2401 aaacccatca gcgagctggc cacctctgtg ctgacccacc gctttccaga ctccacctcc
     2461 tcacccggcg aacatggcct tggacgaatg cagttgtcga tccgctacag cgcccagcgt
     2521 caaaaactag acgtgaccat acacaaaatc cagaagatac cacttcgcga tcccagcaat
     2581 atccccgatc cgtatgttaa gctgtatctg ttgcctggac gcaccaagga gtcgaaacgc
     2641 aagacgagcg tgatcaagga caactgcaac cccgtctacg atgcatcctt tgagtacctg
     2701 atttccattg ccgaactcag gcagacggaa ctggaggtga cggtgtgcac ccaaaaggga
     2761 ttcctatccg gcggtagtcc catcattggc atggtaggta cccgaaagca accccttagt
     2821 tacagacaca gcgcgtacgt ccttcgcatc cttatgattc ccaagtacat attctgcaag
     2881 agtacagtat atataggaaa gatatccggg tgaacttcgc cacaattgtc gatcgtcagt
     2941 tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg
     3001 caccgagtcg gtgctttttt gctcacctgt gattgctcct actcaaatac aaaaacatca
     3061 aattttctgt caataaagca tatttattta tatttatttt acaggaaaga attcctttta
     3121 aagtgtattt taacctataa tgaaaaacga ttaaaaaaaa tacataaaat aattcgaaaa
     3181 tttttgaata gcccaggttg ataaaaattc atttcatacg ttttataact tatgccccta
     3241 agtatttttt gaccatagtg tttcaattct acattaattt tacagagtag aatgaaacgc
     3301 cacctactca gccaagaggc gaaaaggtta gctcgccaag cagagagggc gccagtgctc
     3361 actacttttt ataattctca acttcttttt ccagactcag ttcgtatata tagacctatt
     3421 ttcaatttaa cgtcggtttt ggacactgga accggtttta gagctagaaa tagcaagtta
     3481 aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc ttttttgcct
     3541 acctggagcc tgagagttgt tcaataaaat aaaaatgttt cgtttttttg ctttcgccag
     3601 tatttattat ttttcatcaa tatgtattca atttggtatg tatttagtaa ttgtaatata
     3661 tagacaatgg ttttccgttg acgtacatac atctgacgtg tgtttattta gacataatag
     3721 ttatgttttc acatcttttt aatgttcgct taatgcgtat gcattctaga caattgtgct
     3781 cggcaacagt atatttgtgg tgtgccaacc aacaacctgc aggagctcca gcttttgtgc
     3841 tagcgtacgc gtatcgataa gctttaagat acattgatga gtttggacaa accacaacta
     3901 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     3961 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     4021 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg
     4081 ctgattatga tctagagtcg cggccgcttt acttgtacag ctcgtccatg ccgagagtga
     4141 tcccggcggc ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt
     4201 tgctcagggc ggactgggtg ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc
     4261 cgatgggggt gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt
     4321 ggcggatctt gaagttcacc ttgatgccgt tcttctgctt gtcggccatg atatagacgt
     4381 tgtggctgtt gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt
     4441 cgatgccctt cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc
     4501 gggtcttgta gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg
     4561 gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg ctgaagcact
     4621 gcacgccgta ggtcagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg
     4681 tgcagatgaa cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca
     4741 cgctgaactt gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg
     4801 tgaacagctc ctcgcccttg ctcaccatgg tggcgaccgg tggatccctg gcccgcggta
     4861 ccgtcgactc tagcggtacc ccgattgttt agcttgttca gctgcgcttg tttatttgct
     4921 tagctttcgc ttagcgacgt gttcactttg cttgtttgaa ttgaattgtc gctccgtaga
     4981 cgaagcgcct ctatttatac tccggcggtc gagggttcga aatcgataag cttggatcct
     5041 aattgaatta gctctaattg aattagtctc taattgaatt agatccccgg gcgagctcgg
     5101 acgtctcaag ggtctgcagc aagacttcgt gcctagagct ctgcaccgca tcttcgagga
     5161 gcagcagctg cggcatgccg acaaggtggc tctgatctat cagcccagca ctacgggcca
     5221 gggaatggcg cccagtcaga gcagctaccg ccagatgaac gagcgtgcga accgggcagc
     5281 ccgcctcctg gtggccgaga cccacggccg gttcctgcag ccaaacagcg atggtgactt
     5341 catcgtggct gtgtgcatgc agccgtcgga gggattggtc accacactgc tggccatctg
     5401 gaaggctggc ggcgcgtatt tgcccatcga tcccagcttc ccggcgaacc gcattcacca
     5461 catactgctg gaggcgaagc ccaccttggt gattcgcgac gatgacatcg acgccggccg
     5521 tttccaggga actcccacgt tatccaccac cgaactgtat gccaaatccc tccagctggc
     5581 cggctccaat ctgctctcag aggagatgct gcgcggtggc aacgaccaca tcgccatcgt
     5641 gctctacacc tcgggcagta cgggtgtgcc caagggagtg cgtctgccgc acgagagcat
     5701 cctcaatcgg ctccagtggc agtgggcgac ctttccgtac accgccaacg aggcggtgag
     5761 cgttttcaag acggccttaa ccttcgtgga ctcgattgcg gagctatggg gcccgttaat
     5821 gtgtggcctg gccattctgg tggtgcccaa ggcagtgacc aaggatcccc agcgattggt
     5881 ggccctgttg gagcgttata agatcaggcg tctcgtactg gtgcccaccc ttctgcgctc
     5941 gttgctcatg tatctgaaga tggagggagg cggagctgct cagaaactgc tgtacaatct
     6001 ccagatttgg gtctgctccg gagaacccct atccgtttcc ctggccagca gcttcttcga
     6061 ctatttcgat gagggcgtgc accggctgta caacttttac ggatccacga agggcgaatt
     6121 ctgcagatat ccatcacact ggcggccgct cgagcatgca tctagagggc ccaattcgcc
     6181 ctatagtgag tcgtattaca attcactggc cgtcgtttta caacgtcgtg actgggaaaa
     6241 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     6301 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     6361 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     6421 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     6481 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     6541 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     6601 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     6661 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     6721 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     6781 aacgcgaatt ttaacaaaat tcagggcgca agggctgcta aaggaagcgg aacacgtaga
     6841 aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt cagctactgg gctatctgga
     6901 caagggaaaa cgcaagcgca aagagaaagc aggtagcttg cagtgggctt acatggcgat
     6961 agctagactg ggcggtttta tggacagcaa gcgaaccgga attgccagct ggggcgccct
     7021 ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttcttgccg ccaaggatct
     7081 gatggcgcag gggatcaaga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     7141 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     7201 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     7261 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     7321 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     7381 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     7441 tgtcatccca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     7501 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     7561 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     7621 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     7681 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     7741 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     7801 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     7861 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     7921 tcttctgaat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     7981 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     8041 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     8101 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     8161 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     8221 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     8281 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     8341 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     8401 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     8461 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     8521 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     8581 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     8641 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     8701 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     8761 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     8821 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     8881 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     8941 agcgtcagac cccgtagaaa agatcaaagg atcttc
//
