LOCUS       Exported                4577 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAV012, complete sequence.
ACCESSION   KF411443
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4577)
  AUTHORS   White KP, Victorsen A.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-JUL-2013) Human Genetics, The University of Chicago, 
            5801 South Ellis Avenue, Chicago, IL 60637, USA
REFERENCE   2  (bases 1 to 4577)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4577
                     /organism="Cloning vector pAV012"
                     /mol_type="other DNA"
                     /db_xref="taxon:1400557"
     rep_origin      26..414
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     rep_origin      41..414
                     /label=R6K ori
                     /note="R6K ori"
     promoter        509..613
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      544..549
                     /regulatory_class="minus_35_signal"
                     /note="bla"
     regulatory      565..570
                     /regulatory_class="minus_10_signal"
                     /note="bla"
     CDS             614..1474
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="ampicillin resistance protein"
                     /protein_id="AGV76757.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             join(1583..1867,3653..4288)
                     /codon_start=1
                     /product="AV12 N-terminal fusion tag protein"
                     /label=AV12 N-terminal fusion tag protein
                     /protein_id="AGV76758.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKESSGSFLNCCPGCCMEPGSGNWSHPQFEKENLYFQSDYKDHD
                     GDYKDHDIDYKDDDDKSALTDDLPRA"
     misc_feature    1583..1867
                     /note="sGFP exon 1; 1st exon of superfolder green 
                     fluorescent protein"
     primer_bind     1583..1604
                     /label=forward recombineering primer binding site
                     /note="forward recombineering primer binding site"
     intron          1868..3652
                     /note="sGFP intron"
     misc_feature    1941..1974
                     /label=loxP
                     /note="loxP"
     protein_bind    1941..1974
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      1984..2491
                     /regulatory_class="promoter"
                     /note="PGK promoter"
     promoter        1989..2488
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     promoter        2505..2552
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     regulatory      2520..2525
                     /regulatory_class="minus_35_signal"
                     /note="gb2 promoter"
     regulatory      2539..2547
                     /regulatory_class="minus_10_signal"
                     /note="gb2 promoter"
     CDS             2571..3374
                     /codon_start=1
                     /product="AphA"
                     /label=AphA
                     /protein_id="AGV76759.1"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPGGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    3507..3540
                     /label=loxP
                     /note="loxP"
     protein_bind    3507..3540
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        3604..3622
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     misc_feature    3653..4192
                     /note="sGFP exon 2; 2nd exon of superfolder green 
                     fluorescent protein"
     CDS             4109..4126
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     CDS             4148..4171
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4172..4192
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     misc_feature    4193..4258
                     /label=3xFLAG epitope
                     /note="3xFLAG epitope"
     CDS             4193..4258
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(4265..4288)
                     /label=reverse recombineering primer binding site
                     /note="reverse recombineering primer binding site"
     oriT            4334..4443
                     /note="incP origin of transfer"