LOCUS       Exported                4232 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pR6Kamp-LAP(GFP), complete sequence.
ACCESSION   MH170480
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4232)
  AUTHORS   Poser I, Sarov M, Hutchins JR, Heriche JK, Toyoda Y, Pozniakovsky A,
            Weigl D, Nitzsche A, Hegemann B, Bird AW, Pelletier L, Kittler R, 
            Hua S, Naumann R, Augsburg M, Sykora MM, Hofemeister H, Zhang Y, 
            Nasmyth K, White KP, Dietzel S, Mechtler K, Durbin R, Stewart AF, 
            Peters JM, Buchholz F, Hyman AA.
  TITLE     BAC TransgeneOmics: a high-throughput method for exploration of 
            protein function in mammals
  JOURNAL   Nat. Methods 5 (5), 409-415 (2008)
  PUBMED    18391959
REFERENCE   2  (bases 1 to 4232)
  AUTHORS   Hyman AA.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-APR-2018) Human Genetics, University of Chicago, 900 E
            57th St, Kcbd 10240, Chicago, IL 60605, USA
REFERENCE   3  (bases 1 to 4232)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4232
                     /organism="Cloning vector pR6Kamp-LAP(GFP)"
                     /mol_type="other DNA"
                     /db_xref="taxon:2306014"
     CDS             1..915
                     /codon_start=1
                     /product="LAP tag fusion protein"
                     /label=LAP tag fusion protein
                     /protein_id="AXR98587.1"
                     /translation="DYDIPTTASENLYFQGAAKFKETAAAKFERQHMDSGGGGSSGPSG
                     SSSLEVLFQGPLSSSGPSGSMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDAT
                     YGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERT
                     IFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADK
                     QKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKR
                     DHMVLKEFVTAAGITLGMDELYK"
     misc_feature    1..27
                     /label=forward recombineering primer sequence
                     /note="forward recombineering primer sequence"
     misc_feature    28..48
                     /label=TEV cleavage site
                     /note="TEV cleavage site"
     CDS             28..48
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     misc_feature    61..105
                     /label=S-peptide
                     /note="S-peptide"
     CDS             61..105
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     misc_feature    145..168
                     /label=precision
                     /note="precision"
     CDS             145..168
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     misc_feature    196..915
                     /label=eGFP
                     /note="eGFP"
     CDS             196..915
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLK
                     EFVTAAGITLGMDELYK"
     misc_feature    953..1538
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     regulatory      954..1525
                     /regulatory_class="ribosome_binding_site"
                     /note="internal ribosomal entry site (IRES)"
     regulatory      1541..1546
                     /regulatory_class="minus_35_signal"
     regulatory      1561..1569
                     /regulatory_class="minus_10_signal"
     CDS             1620..2423
                     /codon_start=1
                     /product="neomycin phosphotransferase II"
                     /label=neomycin phosphotransferase II
                     /note="kanamycin and neomycin resistance"
                     /protein_id="AXR98588.1"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    complement(2402..2423)
                     /label=reverse recombineering primer sequence
                     /note="reverse recombineering primer sequence"
     oriT            2457..2566
                     /note="incP origin of transfer"
     rep_origin      2725..3113
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     rep_origin      2740..3113
                     /label=R6K ori
                     /note="R6K ori"
     promoter        3207..3311
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      3242..3247
                     /regulatory_class="minus_35_signal"
     regulatory      3263..3268
                     /regulatory_class="minus_10_signal"
     CDS             3312..4172
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="ampicillin resistance"
                     /protein_id="AXR98589.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNVGDHVTRLDRW
                     EPELNEAIPNDERDATMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMGERNRQIAEIGAS
                     LIKHW"