LOCUS       62056_22575        4862 bp DNA     circular SYN 07-OCT-2020
DEFINITION  Cloning vector pVG363_Aste-U6a-Swap4-gRNA, complete sequence.
ACCESSION   MW030450
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4862)
  AUTHORS   Adolfi A, Gantz V, Jasinskiene N, Lee H-F., Hwang K, Terradas G, 
            Bulger E, Ramaiah A, Bennett J, Emerson JJ, Marshall J, Bier E.
  TITLE     Efficient population modification gene-drive rescue system in the 
            malaria mosquito Anopheles stephensi
  JOURNAL   Nat Commun (2020) In press
REFERENCE   2  (bases 1 to 4862)
  AUTHORS   Adolfi A, Gantz V, Jasinskiene N, Lee H-F., Hwang K, Terradas G, 
            Bulger E, Ramaiah A, Bennett J, Emerson JJ, Marshall J, Bier E.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-SEP-2020) Developmental and Stem Cell Biology, 
            University of California, San Francisco, 1650 Owens St., San 
            Francisco, CA 94158, USA
REFERENCE   3  (bases 1 to 4862)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Commun 
            (2020) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (22-SEP-2020) Developmental and Stem Cell Biology, University of 
            California, San Francisco, 1650 Owens St., San Francisco, CA 94158, 
            USA"
FEATURES             Location/Qualifiers
     source          1..4862
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(333..1124)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      complement(1468..1896)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     2037..2053
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2060..2078
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        2597..2672
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(3153..3169)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3177..3193)
                     /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(3201..3231)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3246..3267)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3555..4143)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(4317..4862,1..312))
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
       61 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
      121 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
      181 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
      241 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
      301 ttgaatactc atactcttcc tttttcaatt cagaagaact cgtcaagaag gcgatagaag
      361 gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat
      421 tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc
      481 gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata
      541 ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catgcgcgcc
      601 ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc
      661 tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg
      721 tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg
      781 atggatactt tctcggcagg agcaaggtgg gatgacagga gatcctgccc cggcacttcg
      841 cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga
      901 acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cctgcagttc attcagggca
      961 ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg
     1021 gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc
     1081 caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat
     1141 cctgtctctt gatcagatct tgatcccctg cgccatcaga tccttggcgg caagaaagcc
     1201 atccagttta ctttgcaggg cttcccaacc ttaccagagg gcgccccagc tggcaattcc
     1261 ggttcgcttg ctgtccataa aaccgcccag tctagctatc gccatgtaag cccactgcaa
     1321 gctacctgct ttctctttgc gcttgcgttt tcccttgtcc agatagccca gtagctgaca
     1381 ttcatccggg gtcagcaccg tttctgcgga ctggctttct acgtgttccg cttcctttag
     1441 cagcccttgc gccctgaatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     1501 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     1561 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     1621 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     1681 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     1741 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     1801 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     1861 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat tcgccattca ggctgcgcaa
     1921 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     1981 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     2041 aacgacggcc agtgaattgt aatacgactc actatagggc gaattgggcc ctctagatgc
     2101 atgctcgagc ggccgccagt gtgatggata tctgcagaat tcgcccttaa gcttatcgat
     2161 acgcgtacct caactgccga cgagttgctc gggtttcgat gagttccagc atacgcgcat
     2221 gctgctgagt gcttcttggt cgcatataca ctcatgccct ttttgccttg tgacgatttt
     2281 tttttttcta ctcaatgcaa atttcaaatt tgagggcaat tttcgtgctt atgtttaata
     2341 atctataaaa gaaaatttaa aactaaaaac tcatgtacag cagtagatga agctttcacc
     2401 gttcttctca cttaaattct acctgcacat tggctggtgg aagagctata attgaagcag
     2461 aaccttataa cagtcgcgcg gaaggatcaa gagctttgtg aatatccttt atccatcgct
     2521 agaactaaaa cggatgacag acgggatata taccaaccca tttgccccct cccttgttac
     2581 gttaattaac gtagaagttt tagagctaga aatagcaagt taaaataagg ctagtccgtt
     2641 atcaacttga aaaagtggca ccgagtcggt gctttttttg tggaaatttg attcacttgt
     2701 tttaggaaaa taattacctt cctttgaaac aggtatttac aatgatggac gatagaaaaa
     2761 gaccacctta aaatgcttta ttcaagctat tgctttcgat tcttcattga gatctaatct
     2821 caaatttgtg tatttagaaa aaacagcgaa tattgtacta taataattct ttggtaatct
     2881 caggctttca gtaatgatag aatccttaag actgtaagaa tatacaagtt ttaaaaaaag
     2941 gaaaacaggt atatcaaatc agaaaacaac aacagctctt ttccgttgaa ttctacctgt
     3001 acgatggctt aaggatgagc tgtaactgaa acgaaacctt acaacagcca cgcagaacgt
     3061 aggccggccg atctcggata agggcgaatt ccagcacact ggcggccgtt actagtggat
     3121 ccgagctcgg taccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     3181 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     3241 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3301 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3361 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3421 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3481 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3541 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3601 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3661 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3721 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3781 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3841 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3901 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3961 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4021 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4081 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4141 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4201 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4261 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4321 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4381 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4441 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4501 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4561 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4621 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4681 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4741 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4801 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4861 gg
//
