LOCUS       62056_22560        8918 bp DNA     circular SYN 02-NOV-2019
DEFINITION  Expression vector pVG307_truncated, complete sequence.
ACCESSION   MN551091
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8918)
  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  JOURNAL   Unpublished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 8918)
  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 8918)
  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            "Unpublished"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..8918
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      611..1199
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    1487..1508
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        1523..1553
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1561..1577
                     /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     1585..1601
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2688..3484
                     /label=dU6-1 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-1
                     snRNA (Port et al., 2014)"
     misc_RNA        3504..3579
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3580..4000
                     /label=dU6-3 promoter
                     /note="RNA polymerase III promoter for Drosophila U6-3
                     snRNA (Port et al., 2014)"
     polyA_signal    complement(4430..4551)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(4677..5393)
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     promoter        complement(6732..6750)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6757..6773)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6914..7342
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             7686..8477
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             join(8498..8918,1..437)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
       61 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
      121 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
      181 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
      241 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
      301 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
      361 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
      421 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
      481 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
      541 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
      601 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
      661 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
      721 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
      781 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
      841 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
      901 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
      961 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     1021 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     1081 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     1141 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     1201 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     1261 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     1321 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     1381 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     1441 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     1501 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     1561 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg
     1621 gtaccgagct cggatccact agtaacggcc gccagtgtgc tggaattcgc ccttgctcaa
     1681 agtccccaaa cgaaagccaa agatgccagc gatttacact cgcaattgtc ttatgaggtg
     1741 gcaacatggt ggaggaacct tgtcactccc accacccatc gtcccatcgt cccatcgtcc
     1801 atcgcccaac gacctgtgac cccggccccg tcaaccgtcc accgcccagc gcccactttc
     1861 aaggccatcc accaagtgcc agccctaatt tgcacatttt aaataggcct cgccatctct
     1921 ttttttggga aacttggaaa actcggatct tgggcatttt tattaaacag atgggctcct
     1981 tctttataaa gccgaggggg tggggtgggg gttagtgccc ggaaacaggg acggcggagc
     2041 agggattagc caggctgggc tagatttatg cacagacgcc ttcatcttta gggacgaacg
     2101 ccgtgaaatt gaaaagtttt cagcttgaaa ccttttctag atgcacaaaa aataaataaa
     2161 agtataaacc tacttcgtag gatacttcgt tttgttcggg gttagatgag cataacgctt
     2221 gtagttgata tttgagatcc cctatcattg cagggtgaca gcggagcggc ttcgcagagc
     2281 tgcattaacc agggcttcgg gcaggccaaa aactacggca cgctcctgcc acccagtccg
     2341 ccggaggact ccggttcagg gagcggccaa ctagccgaga acctcaccta tgcctggcac
     2401 aatatggaca tctttggggc ggtcaatcag ccgggctccg gatggcggca gctggtcaac
     2461 cggacacgcg gactattctg caacgagcga cacataccgg cgcccaggaa acatttgctc
     2521 aagaacggtg agtttctatt cgcagtcggc tgatctgtgt gaaatcttaa taaagggtcc
     2581 aattaccaat ttgaaactca gtttgcggcg tggcctatcc gggcgaactt ttggccgtga
     2641 tgggcagttc cggtgccgga aagacgaccc tgctgaatgc ccacgtcatt ttcaacgtcc
     2701 tcgatagtat agtggttagt atccccgcct gtcacgcggg agaccggggt tcaattcccc
     2761 gtcggggaga atctgtgatt cttttttttt ttcttttact ttgttatata aacaattttt
     2821 gttttaattg aatctaattt gccattgctt ttaggaatct caggcatcca gcaagcgttt
     2881 gtccgccgaa tcgcccatca gtgaagaaga tcctgtggcg gctacgaaaa tctccccggc
     2941 catgtcggcc tccacctcca gcgaaaaacc catcagcgag ctggccacct ctgtgctgac
     3001 ccaccgcttt ccagactcca cctcctcacc cggcgaacat ggccttggac gaatgcagtt
     3061 gtcgatccgc tacagcgccc agcgtcaaaa actagacgtg accatacaca aaatccagaa
     3121 gataccactt cgcgatccca gcaatatccc cgatccgtat gttaagctgt atctgttgcc
     3181 tggacgcacc aaggagtcga aacgcaagac gagcgtgatc aaggacaact gcaaccccgt
     3241 ctacgatgca tcctttgagt acctgatttc cattgccgaa ctcaggcaga cggaactgga
     3301 ggtgacggtg tgcacccaaa agggattcct atccggcggt agtcccatca ttggcatggt
     3361 aggtacccga aagcaacccc ttagttacag acacagcgcg tacgtccttc gcatccttat
     3421 gattcccaag tacatattct gcaagagtac agtatatata ggaaagatat ccgggtgaac
     3481 ttcggcgata cttggatgcc ctggttttag agctagaaat agcaagttaa aataaggcta
     3541 gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttttgctca cctgtgattg
     3601 ctcctactca aatacaaaaa catcaaattt tctgtcaata aagcatattt atttatattt
     3661 attttacagg aaagaattcc ttttaaagtg tattttaacc tataatgaaa aacgattaaa
     3721 aaaaatacat aaaataattc gaaaattttt gaatagccca ggttgataaa aattcatttc
     3781 atacgtttta taacttatgc ccctaagtat tttttgacca tagtgtttca attctacatt
     3841 aattttacag agtagaatga aacgccacct actcagccaa gaggcgaaaa ggttagctcg
     3901 ccaagcagag agggcgccag tgctcactac tttttataat tctcaacttc tttttccaga
     3961 ctcagttcgt atatatagac ctattttcaa tttaacgtcg gttttggaca ctggaaccgg
     4021 ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg
     4081 gcaccgagtc ggtgcttttt tgcctacctg gagcctgaga gttgttcaat aaaataaaaa
     4141 tgtttcgttt ttttgctttc gccagtattt attatttttc atcaatatgt attcaatttg
     4201 gtatgtattt agtaattgta atatatagac aatggttttc cgttgacgta catacatctg
     4261 acgtgtgttt atttagacat aatagttatg ttttcacatc tttttaatgt tcgcttaatg
     4321 cgtatgcatt ctagacaatt gtgctcggca acagtatatt tgtggtgtgc caaccaacaa
     4381 cctgcaggag ctccagcttt tgtgctagcg tacgcgtatc gataagcttt aagatacatt
     4441 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     4501 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     4561 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
     4621 taaaacctct acaaatgtgg tatggctgat tatgatctag agtcgcggcc gctttacttg
     4681 tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag caggaccatg
     4741 tgatcgcgct tctcgttggg gtctttgctc agggcggact gggtgctcag gtagtggttg
     4801 tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc
     4861 tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt tcaccttgat gccgttcttc
     4921 tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc
     4981 aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt caccagggtg
     5041 tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg
     5101 gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg
     5161 tggtcggggt agcggctgaa gcactgcacg ccgtaggtca gggtggtcac gagggtgggc
     5221 cagggcacgg gcagcttgcc ggtggtgcag atgaacttca gggtcagctt gccgtaggtg
     5281 gcatcgccct cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc gccgtccagc
     5341 tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac catggtggcg
     5401 accggtggat cccgggcccg cggtaccgtc gactctagcg gtaccccgat tgtttagctt
     5461 gttcagctgc gcttgtttat ttgcttagct ttcgcttagc gacgtgttca ctttgcttgt
     5521 ttgaattgaa ttgtcgctcc gtagacgaag cgcctctatt tatactccgg cggtcgaggg
     5581 ttcgaaatcg ataagcttgg atcctaattg aattagctct aattgaatta gtctctaatt
     5641 gaattagatc cccgggcgag ctcgcatatc gggatgcgac tgctcaatgg ccaacctgtg
     5701 gacgccaagg agatgcaggc caggtgcgcc tatgtccagc aggatgacct ctttatcggc
     5761 tccctaacgg ccagggagca cctgattttc caggccatgg tgcggatgcc acgacatctg
     5821 acctatcggc agcgagtggc ccgcgtggat caggtgatcc aggagctttc gctcagcaaa
     5881 tgtcagcaca cgatcatcgg tgtgcccggc agggtgaaag gtctgtccgg cggagaaagg
     5941 aagcgtctgg cattcgcctc cgaggcacta accgatccgc cgcttctgat ctgcgatgag
     6001 cccacctccg gactggactc atttaccgcc cacagcgtcg tccaggtgct gaagaagctg
     6061 tcgcagaagg gcaagaccgt catcctgacc attcatcagc cgtcttccga gctgtttgag
     6121 ctctttgaca agatccttct gatggccgag ggcagggtag ctttcttggg cactcccagc
     6181 gaagccgtcg acttcttttc ctagtgagtt cgatgtgttt attaagggta tctagcatta
     6241 cattacatct caactcctat ccagcgtggg tgcccagtgt cctaccaact acaatccggc
     6301 ggacttttac gtacaggtgt tggccgttgt gcccggacgg gagatcgagt cccgtgatcg
     6361 gatcgccaag atatgcgaca attttgccat tagcaaagta gcccgggata tggagcagtt
     6421 gttggccacc aaaaatttgg agaagccact ggagcagccg gagaacgggt acacctacaa
     6481 ggccacctgg ttcatgcagt tccgggcggt cctgtggcga tcctggctgt cggtgctcaa
     6541 ggaaccactc ctcgtaaaag tgcgacttat tcagacaacg gtgagtggtt ccagtggaaa
     6601 caaatgatat aacgcttaca attcttggaa acaaattcgc tagattttag atagaattgc
     6661 caagggcgaa ttctgcagat atccatcaca ctggcggccg ctcgagcatg catctagagg
     6721 gcccaattcg ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg
     6781 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     6841 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     6901 gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     6961 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     7021 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     7081 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     7141 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     7201 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     7261 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     7321 taacaaaaat ttaacgcgaa ttttaacaaa attcagggcg caagggctgc taaaggaagc
     7381 ggaacacgta gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat gtcagctact
     7441 gggctatctg gacaagggaa aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc
     7501 ttacatggcg atagctagac tgggcggttt tatggacagc aagcgaaccg gaattgccag
     7561 ctggggcgcc ctctggtaag gttgggaagc cctgcaaagt aaactggatg gctttcttgc
     7621 cgccaaggat ctgatggcgc aggggatcaa gatctgatca agagacagga tgaggatcgt
     7681 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     7741 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     7801 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     7861 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     7921 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     7981 ggcaggatct cctgtcatcc caccttgctc ctgccgagaa agtatccatc atggctgatg
     8041 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     8101 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     8161 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     8221 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     8281 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     8341 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     8401 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     8461 ttcttgacga gttcttctga attgaaaaag gaagagtatg agtattcaac atttccgtgt
     8521 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     8581 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     8641 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     8701 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     8761 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     8821 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     8881 tgataacact gcggccaact tacttctgac aacgatcg
//
