LOCUS       62056_22550        8905 bp DNA     circular SYN 02-NOV-2019
DEFINITION  Expression vector pVG186_y1-w2, complete sequence.
ACCESSION   MN551088
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8905)
  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  JOURNAL   Unpublished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 8905)
  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 8905)
  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            "Unpublished"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..8905
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(174..1031)
                     /label=AmpR
                     /note="beta-lactamase"
     CDS             complement(1052..1843)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(2187..2615)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     2756..2772
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2779..2797
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2868..3891
                     /label=yellow homology arm
                     /note="yellow homology arm"
     promoter        3898..4694
                     /label=dU6-1 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-1
                     snRNA (Port et al., 2014)"
     misc_RNA        4714..4789
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4790..5210
                     /label=dU6-3 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-3
                     snRNA (Port et al., 2014)"
     misc_feature    5210..5229
                     /label=y1 gRNA
                     /note="y1 gRNA"
     misc_feature    5230..5305
                     /label=guide RNA scaffold for the CRISPR/Cas9 system
                     /note="guide RNA scaffold for the CRISPR/Cas9 system"
     misc_feature    5312..5613
                     /label=U6-3 3' Sequence
                     /note="U6-3 3' Sequence"
     polyA_signal    complement(5640..5761)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(5885..6559)
                     /label=DsRed-Express
                     /note="rapidly maturing tetrameric variant of DsRed
                     fluorescent protein (Bevis and Glick, 2002)"
     misc_feature    complement(6581..6705)
                     /label=Hsp70 Promoter
                     /note="Hsp70 Promoter"
     misc_feature    complement(6737..6780)
                     /label=3xP3 eye promoter
                     /note="3xP3 eye promoter"
     CDS             7077..7088
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
     promoter        complement(7879..7897)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(7915..7931)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(7939..7955)
                     /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(7963..7993)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(8008..8029)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(8317..8905)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaattc agaagaactc gtcaagaagg cgatagaagg
     1081 cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt
     1141 cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg
     1201 ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat
     1261 tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgcgcgcct
     1321 tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct
     1381 gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt
     1441 ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga
     1501 tggatacttt ctcggcagga gcaaggtggg atgacaggag atcctgcccc ggcacttcgc
     1561 ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa
     1621 cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ctgcagttca ttcagggcac
     1681 cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg
     1741 cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc
     1801 aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc
     1861 ctgtctcttg atcagatctt gatcccctgc gccatcagat ccttggcggc aagaaagcca
     1921 tccagtttac tttgcagggc ttcccaacct taccagaggg cgccccagct ggcaattccg
     1981 gttcgcttgc tgtccataaa accgcccagt ctagctatcg ccatgtaagc ccactgcaag
     2041 ctacctgctt tctctttgcg cttgcgtttt cccttgtcca gatagcccag tagctgacat
     2101 tcatccgggg tcagcaccgt ttctgcggac tggctttcta cgtgttccgc ttcctttagc
     2161 agcccttgcg ccctgaattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     2221 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     2281 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     2341 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     2401 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     2461 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     2521 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     2581 acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt cgccattcag gctgcgcaac
     2641 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     2701 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     2761 acgacggcca gtgaattgta atacgactca ctatagggcg aattgggccc tctagatgca
     2821 tgctcgagcg gccgccagtg tgatggatat ctgcagaatt cgcccttgtt gcgaggtttt
     2881 aggactgaaa gagcacatgt caaaatataa atttgttcaa atactttata tttgactgaa
     2941 ttagattgtt attttaaaag ttatgaatta aataaagatt gaaaggtgca ttatgctcaa
     3001 atgtatattt atcgcaaccc ccggttactt tgtaaagcaa aaacgcctgg tttgattttt
     3061 aagaagatgg gtcggtaaat cgataaaagc tatattttct ggtcgttgca gtctcactcg
     3121 cctgctataa aaacattaaa agttcccaga aacaataaat gtctttaaat tcaattaacg
     3181 aagaaataaa gaaggaaaag aactggagcg gaaatcggtc gaaatactgc caatggccac
     3241 atatacattt aacagcgata tatggtatac atattgataa tgatgtcaga cgcaattgct
     3301 tcagacggct aatgacatcg caaattgcac gcaacttgca atagtgccaa ttatgactga
     3361 agtacatata gccggggatc ttttaacata aacttccagt agatgtacaa gcagaaaaaa
     3421 gagccattag cacggcagtt accattgctt atgattcctt gtgtccaaaa taatgacaaa
     3481 taggtatata aataattaaa tgccaaacat aagcgattct aatttacctt tacatctgta
     3541 tgcatttaca tattatccag aaaacagaca gcgataactt gcaacattgc ttagtataat
     3601 aatccaaaga aggaatttag gcagaaattc cagttaatta aatattcaaa acaaacttta
     3661 tttagtgcct caataatagt ttggccctgc taattctcct attttatttt ttagggattc
     3721 cggccactct gacctatata aacatggacc gcagtttgac gggttcaccg gagctaattc
     3781 cgtatccaga ttggcgctca aatacagctg gagattgcgc caacagtatt accactgcct
     3841 accgcattaa agtggatgag tgtggtcggc tgtgggtttt ggacactgga agacgtcatt
     3901 ttcaacgtcc tcgatagtat agtggttagt atccccgcct gtcacgcggg agaccggggt
     3961 tcaattcccc gtcggggaga atctgtgatt cttttttttt ttcttttact ttgttatata
     4021 aacaattttt gttttaattg aatctaattt gccattgctt ttaggaatct caggcatcca
     4081 gcaagcgttt gtccgccgaa tcgcccatca gtgaagaaga tcctgtggcg gctacgaaaa
     4141 tctccccggc catgtcggcc tccacctcca gcgaaaaacc catcagcgag ctggccacct
     4201 ctgtgctgac ccaccgcttt ccagactcca cctcctcacc cggcgaacat ggccttggac
     4261 gaatgcagtt gtcgatccgc tacagcgccc agcgtcaaaa actagacgtg accatacaca
     4321 aaatccagaa gataccactt cgcgatccca gcaatatccc cgatccgtat gttaagctgt
     4381 atctgttgcc tggacgcacc aaggagtcga aacgcaagac gagcgtgatc aaggacaact
     4441 gcaaccccgt ctacgatgca tcctttgagt acctgatttc cattgccgaa ctcaggcaga
     4501 cggaactgga ggtgacggtg tgcacccaaa agggattcct atccggcggt agtcccatca
     4561 ttggcatggt aggtacccga aagcaacccc ttagttacag acacagcgcg tacgtccttc
     4621 gcatccttat gattcccaag tacatattct gcaagagtac agtatatata ggaaagatat
     4681 ccgggtgaac ttcggcgata cttggatgcc ctggttttag agctagaaat agcaagttaa
     4741 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttttgctca
     4801 cctgtgattg ctcctactca aatacaaaaa catcaaattt tctgtcaata aagcatattt
     4861 atttatattt attttacagg aaagaattcc ttttaaagtg tattttaacc tataatgaaa
     4921 aacgattaaa aaaaatacat aaaataattc gaaaattttt gaatagccca ggttgataaa
     4981 aattcatttc atacgtttta taacttatgc ccctaagtat tttttgacca tagtgtttca
     5041 attctacatt aattttacag agtagaatga aacgccacct actcagccaa gaggcgaaaa
     5101 ggttagctcg ccaagcagag agggcgccag tgctcactac tttttataat tctcaacttc
     5161 tttttccaga ctcagttcgt atatatagac ctattttcaa tttaacgtcg gttttggaca
     5221 ctggaaccgg ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact
     5281 tgaaaaagtg gcaccgagtc ggtgcttttt tgcctacctg gagcctgaga gttgttcaat
     5341 aaaataaaaa tgtttcgttt ttttgctttc gccagtattt attatttttc atcaatatgt
     5401 attcaatttg gtatgtattt agtaattgta atatatagac aatggttttc cgttgacgta
     5461 catacatctg acgtgtgttt atttagacat aatagttatg ttttcacatc tttttaatgt
     5521 tcgcttaatg cgtatgcatt ctagacaatt gtgctcggca acagtatatt tgtggtgtgc
     5581 caaccaacaa cctgcaggag ctccagcttt tgtgctagcg tacgcgtatc gataagcttt
     5641 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     5701 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     5761 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     5821 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatctag agtcgcggcc
     5881 cctacaggaa caggtggtgg cggccctcgg cgcgctcgta ctgctccacg atggtgtagt
     5941 cctcgttgtg ggaggtgatg tccagcttgg agtccacgta gtagtagccg ggcagctgca
     6001 cgggcttctt ggccatgtag atggacttga actccaccag gtagtggccg ccgtccttca
     6061 gcttcagggc cttgtggatc tcgcccttca gcacgccgtc gcgggggtac aggcgctcgg
     6121 tggacgcctc ccagcccata gtcttcttct gcattacggg gccgtcggag gggaagttca
     6181 cgccgatgaa cttcaccttg tagatgaagg agccgtcctg gagggaggag tcctgggtca
     6241 cggtcaccac gccgccgtcc tcgaagttca tcacgcgctc ccacttgaag ccctcgggga
     6301 aggacagctt cttgtagtcg gggatgtcgg cggggtgctt cacgtacacc ttggagccgt
     6361 actggaactg gggggacagg atgtcccagg cgaagggcag ggggccgccc ttggtcacct
     6421 tcagcttggc ggtctgggtg ccctcgtagg ggcggccctc gccctcgccc tcgatctcga
     6481 actcgtggcc gttcacggag ccctccatgc gcaccttgaa gcgcatgaac tccttgatga
     6541 cgtcctcgga ggaggccatg gtggcgaccg gcttcgagcc gattgtttag cttgttcagc
     6601 tgcgcttgtt tatttgctta gctttcgctt agcgacgtgt tcactttgct tgtttgaatt
     6661 gaattgtcgc tccgtagacg aagcgcctct atttatactc cggcggtcga gggttcgaaa
     6721 tcgataagct tggatcctaa ttgaattagc tctaattgaa ttagtctcta attgaattag
     6781 atccccgggc gagctcggtc gaccgtgggc atcggcaata ccaccactaa tccgtgcccc
     6841 tatgcggtaa atgtctttga cttgaccacg gatacgcgaa ttcggagata cgagctacct
     6901 ggcgtggaca caaatccaaa tactttcata gctaacattg ccgtggatat aggcaaaaat
     6961 tgcgatgatg catatgccta ttttgccgat gaattgggat acggcttgat tgcttactcc
     7021 tgggaactga acaagtcctg gagattctcg gcacattcgt attttttccc cgatccattg
     7081 aggggcgatt tcaatgtcgc tggtattaac ttccaatggg gcgaggaggg tatatttggt
     7141 atgtcccttt cgcccattcg atcggatggt tatcgtaccc tgtactttag tccgttagca
     7201 agtcatcgac aatttgccgt atccacgagg attttgaggg atgaaaccag gacggaagat
     7261 agctatcatg actttgttgc cttagatgaa cggggtccaa actcccatac cacttcacgt
     7321 gtgatgagcg atgatggaat tgagctgttc aatttaatag atcaaaatgc agtgggttgc
     7381 tggcactcat caatgccgta ctcaccgcaa tttcatggca ttgtggatcg cgatgacgtt
     7441 ggcttagttt ttccggccga tgtgaaaatt gatgagaaca aaaacgtttg ggttctatcc
     7501 gataggatgc ccgttttctt gctgtctgac ttggattatt cagatactaa tttccgaatt
     7561 tacacggctc ccttggccac tttaattgag aatactgtgt gtgatttgag gaataacgcc
     7621 tatgggccgc caaataccgt ttcaatacca aaacaagccg ttttgccaat gggtccaccg
     7681 ttatatacga aacaatatcg tcctgtcttg ccacagaaac ctcagaccag ctgggcttcc
     7741 tcgccgcctc ctccaagtcg cacttatttg cccgccaatt caggcaatgt agtctccaga
     7801 agggcgaatt ccagcacact ggcggccgtt actagtggat ccgagctcgg taccaagctt
     7861 gatgcatagc ttgagtattc tatagtgtca cctaaatagc ttggcgtaat catggtcata
     7921 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     7981 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     8041 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     8101 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     8161 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     8221 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     8281 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     8341 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     8401 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     8461 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     8521 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     8581 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     8641 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     8701 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     8761 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     8821 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     8881 tacgcgcaga aaaaaaggat ctcaa
//
