LOCUS       Exported                7032 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCRII-tra4R-3xP3CFP-tra4L, complete sequence.
ACCESSION   KY171964
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7032)
  AUTHORS   Karaminejadranjbar M, Eckermann KN, Ahmed HMM., Dippel S, Wimmer EA.
  TITLE     Instant induction of resistance evolution in CRISPR/Cas9 gene drive
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7032)
  AUTHORS   Karaminejadranjbar M, Eckermann KN, Ahmed HMM., Dippel S, Wimmer EA.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-NOV-2016) Developmental Biology, 
            Georg-August-University Goettingen, Justus-von-Liebig-Weg 11, 
            Goettingen 37077, Germany
REFERENCE   3  (bases 1 to 7032)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: Geneious v. 8.0.5
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7032
                     /organism="Cloning vector pCRII-tra4R-3xP3CFP-tra4L"
                     /mol_type="other DNA"
                     /db_xref="taxon:2044871"
     misc_feature    16..1033
                     /label=left homologous arm tra g4
                     /note="left homologous arm tra g4"
     protein_bind    1034..1083
                     /label=attP binding site
                     /bound_moiety="attP"
     regulatory      complement(1088..1310)
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40"
     polyA_signal    1117..1198
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(1321..2040)
                     /codon_start=1
                     /product="ECFP"
                     /label=ECFP
                     /protein_id="ATN29912.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      2037..2046
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     regulatory      complement(2194..2282)
                     /regulatory_class="promoter"
                     /note="3xP3"
     protein_bind    complement(2288..2337)
                     /label=attP binding site
                     /bound_moiety="attP"
     misc_feature    2339..3207
                     /label=right homologous Arm tra g4
                     /note="right homologous Arm tra g4"
     promoter        complement(3201..3219)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3226..3242)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3383..3811
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             4155..4949
                     /codon_start=1
                     /product="NeoR/KanR"
                     /label=NeoR/KanR
                     /protein_id="ATN29913.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4967..5827
                     /codon_start=1
                     /product="AmpR"
                     /label=AmpR
                     /protein_id="ATN29914.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5982..6601
                     /label=pBR322
                     /note="pBR322"
     rep_origin      5998..6586
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6874..6895
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6910..6940
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6948..6964
                     /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)."
     primer_bind     6972..6988
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7006..7024
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"