LOCUS       Exported               11424 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  PiggyBac cloning vector pHomeD-B2-SCE-R, complete sequence.
ACCESSION   HQ159399
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11424)
  AUTHORS   Windbichler N, Menichelli M, Papathanos PA, Thyme SB, Li H, Ulge UY,
            Hovde BT, Baker D, Monnat RJ Jr., Burt A, Crisanti A.
  TITLE     A synthetic homing endonuclease-based gene drive system in the human
            malaria mosquito
  JOURNAL   Nature 473 (7346), 212-215 (2011)
  PUBMED    21508956
REFERENCE   2  (bases 1 to 11424)
  AUTHORS   Windbichler N.
  TITLE     Homing
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 11424)
  AUTHORS   Windbichler N.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-AUG-2010) Imperial College London, Imperial College 
            Road, London SW7 2AZ, England
REFERENCE   4  (bases 1 to 11424)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11424
                     /organism="piggyBac cloning vector pHomeD-B2-SCE-R"
                     /mol_type="other DNA"
                     /db_xref="taxon:935898"
     regulatory      complement(18..440)
                     /regulatory_class="terminator"
                     /gene="beta-2-tubulin"
     gene            complement(18..440)
                     /gene="beta-2-tubulin"
                     /label=beta-2-tubulin
     CDS             complement(456..1286)
                     /codon_start=1
                     /gene="I-sceI"
                     /product="I-SceI"
                     /label=I-sceI
                     /protein_id="ADT78592.1"
                     /translation="MGSSSDDEATADAQHAAPPKKKRKVEDPRFMYPYDVPDYAGMKNI
                     KKNQVMNLGPNSKLLKEYKSQLIELNIEQFEAGIGLILGDAYIRSRDEGKTYCMQFEWK
                     NKAYMDHVCLLYDQWVLSPPHKKERVNHLGNLVITWGAQTFKHQAFNKLANLFIVNNKK
                     TIPNNLVENYLTPMSLAYWFMDDGGKWDYNKNSTNKSIVLNTQSFTFEEVEYLVKGLRN
                     KFQLNCYVKINKNKPIIYIDSMSYLIFYNLIKPYLIPQMMYKLPNTISSETFLK"
     gene            complement(456..1286)
                     /gene="I-sceI"
                     /label=I-sceI
     CDS             complement(1167..1193)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(1212..1232)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      complement(1289..1880)
                     /regulatory_class="promoter"
                     /gene="beta-2-tubulin"
     gene            complement(1289..1880)
                     /gene="beta-2-tubulin"
                     /label=beta-2-tubulin
     misc_feature    1889..2632
                     /label=disrupted GFP
                     /note="disrupted GFP"
     CDS             1903..2622
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    2741..2862
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      2902..5511
                     /regulatory_class="promoter"
                     /gene="actin 5C"
     gene            2902..5511
                     /gene="actin 5C"
                     /label=actin 5C
     promoter        2930..5507
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             5519..6199
                     /codon_start=1
                     /product="RFP"
                     /label=RFP
                     /protein_id="ADT78593.1"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLFL"
     polyA_signal    6367..6797
                     /label=hsp70 poly(A)
                     /note="polyadenlyation signal from the Drosophila 
                     melanogaster hsp70Ab gene"
     protein_bind    7917..7986
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(8022..8038)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8046..8062
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(8070..8100)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8115..8136
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(8424..9012)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(9183..10043)
                     /codon_start=1
                     /gene="ampR"
                     /product="AmpR"
                     /label=ampR
                     /protein_id="ADT78594.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(9183..10043)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(10044..10148)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      11124..11385
                     /regulatory_class="promoter"
                     /gene="3xP3"
     gene            11124..11385
                     /gene="3xP3"
                     /label=3xP3