LOCUS       Exported                7279 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pJL71-2W-gpmcherry, complete sequence.
ACCESSION   KP065813
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7279)
  AUTHORS   Scharschmidt TC, Rosenblum MD.
  TITLE     A wave of regulatory T cells into neonatal skin mediates tolerance 
            to commensal microbes
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7279)
  AUTHORS   Scharschmidt TC, Rosenblum MD.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-OCT-2014) Dermatology, University of California San 
            Francisco, 513 Parnassus Ave, San Francisco, CA 94143, USA
REFERENCE   3  (bases 1 to 7279)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7279
                     /organism="Cloning vector pJL71-2W-gpmcherry"
                     /mol_type="other DNA"
                     /PCR_primers="fwd_name: ts083, fwd_seq: 
                     ggatccgaattcttaggaggatgattatttatggaagcatggggagctttagcaaatt 
                     gggcagttgattcagctggttcaggttcaggttcaggtgtgagcaagggcgaggagga 
                     taatatgg, rev_name: ts084, rev_seq: 
                     ggatccggcgcgccttacttgtacagctcgtccatgccgcctgtagaatgtc"
                     /db_xref="taxon:1676628"
     regulatory      7..306
                     /regulatory_class="terminator"
     CDS             2837..3571
                     /codon_start=1
                     /product="ErmC"
                     /label=ErmC
                     /note="erythromycin resistance cassette"
                     /protein_id="AKT95051.1"
                     /translation="MNEKNIKHSQNFITSKHNIDKIMTNIRLNEHDNIFEIGSGKGHFT
                     LELVKRCNFVTAIEIDHKLCKTTENKLVDHDNFQVLNKDILQFKFPKNQSYKIYGNIPY
                     NISTDIIRKIVFDSIANEIYLIVEYGFAKRLLNTKRSLALLLMAEVDISILSMVPREYF
                     HPKPKVNSSLIRLSRKKSRISHKDKQKYNYFVMKWVNKEYKKIFTKNQFNNSLKHAGID
                     DLNNISFEQFLSLFNSYKLFNK"
     rep_origin      3685..5790
     promoter        3936..4040
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4041..4901
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             4239..4901
                     /codon_start=1
                     /product="Amp"
                     /label=Amp
                     /note="ampicillin resistance"
                     /protein_id="AKT95052.1"
                     /translation="MSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHL
                     TDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEA
                     IPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFI
                     ADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     rep_origin      5072..5660
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5761..6165)
                     /codon_start=1
                     /product="ArgB"
                     /label=ArgB
                     /protein_id="AKT95053.1"
                     /translation="MNYFDNKIDQFATYLQKRNNLDHIQFLQVRLGMQVLAKNIGKLIV
                     MYTIAYILNIFLFTLITNLTFYLIRRHAHGAHAPSSFWCYVESIILFILLPLVIVNFHI
                     NFLIMIILTVISLGVISVYAPSGRQLTQRR"
     regulatory      5792..6479
                     /regulatory_class="promoter"
                     /gene="agr"
     gene            5792..6479
                     /gene="agr"
                     /label=agr
     regulatory      complement(6193..6198)
                     /regulatory_class="minus_10_signal"
                     /gene="agr_P2"
     gene            complement(6193..6198)
                     /gene="agr_P2"
                     /label=agr_P2
     regulatory      complement(6216..6221)
                     /regulatory_class="minus_35_signal"
                     /gene="agr-P2"
     gene            complement(6216..6221)
                     /gene="agr-P2"
                     /label=agr-P2
     gene            6389..6417
                     /gene="agr-P3"
                     /label=agr-P3
     regulatory      6389..6394
                     /regulatory_class="minus_35_signal"
                     /gene="agr-P3"
     regulatory      6412..6417
                     /regulatory_class="minus_10_signal"
                     /gene="agr-P3"
     CDS             6504..7277
                     /codon_start=1
                     /product="2W-gpmcherry"
                     /label=2W-gpmcherry
                     /note="2W antigen + fluorescent protein fusion protein"
                     /protein_id="AKT95054.1"
                     /translation="MEAWGALANWAVDSAGSGSGSGVSKGEEDNMAIIKEFMRFKVHME
                     GSVNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPAD
                     IPDYLKLSFPEGFKWERVMNFEDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQ
                     KKTMGWEASSERMYPEDGALKGEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVN
                     IKLDITSHNEDYTIVEQYERAEGRHSTGGMDELYK"