LOCUS       62056_22645        8938 bp DNA     circular SYN 02-NOV-2019
DEFINITION  Expression vector pVMG48_truncated-R, complete sequence.
ACCESSION   MN551093
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8938)
  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  JOURNAL   Unpublished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 8938)
  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 8938)
  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            "Unpublished"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..8938
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    857..878
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        893..923
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    931..947
                     /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     955..971
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1045..2072
                     /label=white homology arm
                     /note="white homology arm"
     promoter        2078..2874
                     /label=dU6-1 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-1
                     snRNA (Port et al., 2014)"
     misc_feature    2874..2893
                     /label=w2 gRNA
                     /note="w2 gRNA"
     misc_RNA        2894..2969
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2970..3390
                     /label=dU6-3 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-3
                     snRNA (Port et al., 2014)"
     misc_feature    3390..3409
                     /label=y1 gRNA
                     /note="y1 gRNA"
     misc_feature    3410..3485
                     /label=gRNA scaffold
                     /note="gRNA scaffold"
     misc_feature    3492..3793
                     /label=U6-3_3'UTR
                     /note="U6-3_3'UTR"
     polyA_signal    complement(3820..3941)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4067..4783)
                     /label=EGFP
                     /note="enhanced GFP"
     regulatory      4838..4962
                     /label=Hsp70 promoter
                     /note="Hsp70 promoter"
                     /regulatory_class="promoter"
     misc_feature    4994..5037
                     /label=3xP3 eye promoter
                     /note="3xP3 eye promoter"
     misc_feature    5060..6051
                     /label=white homology arm truncated
                     /note="white homology arm truncated"
     promoter        complement(6122..6140)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6147..6163)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6304..6732
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             7076..7867
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             7888..8745
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      8919..8938
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
       61 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
      121 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
      181 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
      241 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
      301 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
      361 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
      421 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
      481 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
      541 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
      601 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
      661 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
      721 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
      781 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
      841 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
      901 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
      961 acagctatga ccatgattac gccaagcttg gtaccgagct cggatccact agtaacggcc
     1021 gccagtgtgc tggaattcgc ccttgctcaa agtccccaaa cgaaagccaa agatgccagc
     1081 gatttacact cgcaattgtc ttatgaggtg gcaacatggt ggaggaacct tgtcactccc
     1141 accacccatc gtcccatcgt cccatcgtcc atcgcccaac gacctgtgac cccggccccg
     1201 tcaaccgtcc accgcccagc gcccactttc aaggccatcc accaagtgcc agccctaatt
     1261 tgcacatttt aaataggcct cgccatctct ttttttggga aacttggaaa actcggatct
     1321 tgggcatttt tattaaacag atgggctcct tctttataaa gccgaggggg tggggtgggg
     1381 gttagtgccc ggaaacaggg acggcggagc agggattagc caggctgggc tagatttatg
     1441 cacagacgcc ttcattttta gggacgaacg ccgtgaaatt gaaaagtttt cagcttgaaa
     1501 ccttttctag atgcacaaaa aataaataaa agtataaacc tacttcgtag gatacttcgt
     1561 tttgttcggg gttagatgag cataacgctt gtagttgata tttgagatcc cctatcattg
     1621 cagggtgaca gcggagcggc ttcgcagagc tgcattaacc agggcttcgg gcaggccaaa
     1681 aactacggca cgctcctgcc acccagtccg ccggaggact ccggttcagg gagcggccaa
     1741 ctagccgaga acctcaccta tgcctggcac aatatggaca tctttggggc ggtcaatcag
     1801 ccgggctccg gatggcggca gctggtcaac cggacacgcg gactattctg caacgagcga
     1861 cacataccgg cgcccaggaa acatttgctc aagaacggtg agtttctatt cgcagtcggc
     1921 tgatctgtgt gaaatcttaa taaagggtcc aattaccaat ttgaaactca gtttgcggcg
     1981 tggcctatcc gggcgaactt ttggccgtga tgggcagttc cggtgccgga aagacgaccc
     2041 tgctgaatgc ccttgccttt cgatcgccgc agacgtcatt ttcaacgtcc tcgatagtat
     2101 agtggttagt atccccgcct gtcacgcggg agaccggggt tcaattcccc gtcggggaga
     2161 atctgtgatt cttttttttt ttcttttact ttgttatata aacaattttt gttttaattg
     2221 aatctaattt gccattgctt ttaggaatct caggcatcca gcaagcgttt gtccgccgaa
     2281 tcgcccatca gtgaagaaga tcctgtggcg gctacgaaaa tctccccggc catgtcggcc
     2341 tccacctcca gcgaaaaacc catcagcgag ctggccacct ctgtgctgac ccaccgcttt
     2401 ccagactcca cctcctcacc cggcgaacat ggccttggac gaatgcagtt gtcgatccgc
     2461 tacagcgccc agcgtcaaaa actagacgtg accatacaca aaatccagaa gataccactt
     2521 cgcgatccca gcaatatccc cgatccgtat gttaagctgt atctgttgcc tggacgcacc
     2581 aaggagtcga aacgcaagac gagcgtgatc aaggacaact gcaaccccgt ctacgatgca
     2641 tcctttgagt acctgatttc cattgccgaa ctcaggcaga cggaactgga ggtgacggtg
     2701 tgcacccaaa agggattcct atccggcggt agtcccatca ttggcatggt aggtacccga
     2761 aagcaacccc ttagttacag acacagcgcg tacgtccttc gcatccttat gattcccaag
     2821 tacatattct gcaagagtac agtatatata ggaaagatat ccgggtgaac ttcggcgata
     2881 cttggatgcc ctggttttag agctagaaat agcaagttaa aataaggcta gtccgttatc
     2941 aacttgaaaa agtggcaccg agtcggtgct tttttgctca cctgtgattg ctcctactca
     3001 aatacaaaaa catcaaattt tctgtcaata aagcatattt atttatattt attttacagg
     3061 aaagaattcc ttttaaagtg tattttaacc tataatgaaa aacgattaaa aaaaatacat
     3121 aaaataattc gaaaattttt gaatagccca ggttgataaa aattcatttc atacgtttta
     3181 taacttatgc ccctaagtat tttttgacca tagtgtttca attctacatt aattttacag
     3241 agtagaatga aacgccacct actcagccaa gaggcgaaaa ggttagctcg ccaagcagag
     3301 agggcgccag tgctcactac tttttataat tctcaacttc tttttccaga ctcagttcgt
     3361 atatatagac ctattttcaa tttaacgtcg gttttggaca ctggaaccgg ttttagagct
     3421 agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc
     3481 ggtgcttttt tgcctacctg gagcctgaga gttgttcaat aaaataaaaa tgtttcgttt
     3541 ttttgctttc gccagtattt attatttttc atcaatatgt attcaatttg gtatgtattt
     3601 agtaattgta atatatagac aatggttttc cgttgacgta catacatctg acgtgtgttt
     3661 atttagacat aatagttatg ttttcacatc tttttaatgt tcgcttaatg cgtatgcatt
     3721 ctagacaatt gtgctcggca acagtatatt tgtggtgtgc caaccaacaa cctgcaggag
     3781 ctccagcttt tgtgctagcg tacgcgtatc gataagcttt aagatacatt gatgagtttg
     3841 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     3901 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     3961 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
     4021 acaaatgtgg tatggctgat tatgatctag agtcgcggcc gctttacttg tacagctcgt
     4081 ccatgccgag agtgatcccg gcggcggtca cgaactccag caggaccatg tgatcgcgct
     4141 tctcgttggg gtctttgctc agggcggact gggtgctcag gtagtggttg tcgggcagca
     4201 gcacggggcc gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc
     4261 cgtcctcgat gttgtggcgg atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg
     4321 ccatgatata gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc aggatgttgc
     4381 cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt caccagggtg tcgccctcga
     4441 acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct
     4501 ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg tggtcggggt
     4561 agcggctgaa gcactgcacg ccgtaggtca gggtggtcac gagggtgggc cagggcacgg
     4621 gcagcttgcc ggtggtgcag atgaacttca gggtcagctt gccgtaggtg gcatcgccct
     4681 cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga
     4741 tgggcaccac cccggtgaac agctcctcgc ccttgctcac catggtggcg accggtggat
     4801 cccgggcccg cggtaccgtc gactctagcg gtaccccgat tgtttagctt gttcagctgc
     4861 gcttgtttat ttgcttagct ttcgcttagc gacgtgttca ctttgcttgt ttgaattgaa
     4921 ttgtcgctcc gtagacgaag cgcctctatt tatactccgg cggtcgaggg ttcgaaatcg
     4981 ataagcttgg atcctaattg aattagctct aattgaatta gtctctaatt gaattagatc
     5041 cccgggcgag ctcgcatatc gggatgcgac tgctcaatgg ccaacctgtg gacgccaagg
     5101 agatgcaggc caggtgcgcc tatgtccagc aggatgacct ctttatcggc tccctaacgg
     5161 ccagggaaca cctgattttc caggccatgg tgcggatgcc acgacatctg acctatcggc
     5221 agcgagtggc ccgcgtggat caggtgatcc aggagctttc gctcagcaaa tgtcagcaca
     5281 cgatcatcgg tgtgcccggc agggtgaaag gtctgtccgg cggagaaagg aagcgtctgg
     5341 cattcgcctc cgaggcacta accgatccgc cgcttctgat ctgcgatgag cccacctccg
     5401 gactggactc atttaccgcc cacagcgtcg tccaggtgct gaagaagctg tcgcagaagg
     5461 gcaagaccgt catcctgacc attcatcagc cgtcttccga gctgtttgag ctctttgaca
     5521 agatccttct gatggccgag ggcagggtag ctttcttggg cactcccagc gaagccgtcg
     5581 acttcttttc ctagtgagtt cgatgtgttt attaagggta tctagcatta cattacatct
     5641 caactcctat ccagcgtggg tgcccagtgt cctaccaact acaatccggc ggacttttac
     5701 gtacaggtgt tggccgttgt gcccggacgg gagatcgagt cccgtgatcg gatcgccaag
     5761 atatgcgaca attttgccat tagcaaagta gcccgggata tggagcagtt gttggccacc
     5821 aaaaatttgg agaagccact ggagcagccg gagaatgggt acacctacaa ggccacctgg
     5881 ttcatgcagt tccgggcggt cctgtggcga tcctggctgt cggtgctcaa ggaaccactc
     5941 ctcgtaaaag tgcgacttat tcagacaacg gtgagtggtt ccagtggaaa caaatgatat
     6001 aacgcttaca attcttggaa acaaattcgc tagattttag atagaattgc caagggcgaa
     6061 ttctgcagat atccatcaca ctggcggccg ctcgagcatg catctagagg gcccaattcg
     6121 ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa
     6181 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     6241 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     6301 tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     6361 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     6421 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     6481 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     6541 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     6601 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     6661 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     6721 ttaacgcgaa ttttaacaaa attcagggcg caagggctgc taaaggaagc ggaacacgta
     6781 gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat gtcagctact gggctatctg
     6841 gacaagggaa aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc ttacatggcg
     6901 atagctagac tgggcggttt tatggacagc aagcgaaccg gaattgccag ctggggcgcc
     6961 ctctggtaag gttgggaagc cctgcaaagt aaactggatg gctttcttgc cgccaaggat
     7021 ctgatggcgc aggggatcaa gatctgatca agagacagga tgaggatcgt ttcgcatgat
     7081 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     7141 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     7201 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     7261 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     7321 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     7381 cctgtcatcc caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     7441 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     7501 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     7561 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     7621 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     7681 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     7741 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     7801 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     7861 gttcttctga attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     7921 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     7981 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     8041 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     8101 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     8161 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     8221 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     8281 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     8341 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     8401 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     8461 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     8521 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     8581 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     8641 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     8701 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     8761 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     8821 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     8881 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgc
//
