LOCUS       Exported                7405 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pPV472, complete sequence.
ACCESSION   KC312931
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7405)
  AUTHORS   Massey HC Jr., Ranjit N, Stoltzfus JD, Lok JB.
  TITLE     Strongyloides stercoralis daf-2 encodes a divergent ortholog of 
            Caenorhabditis elegans DAF-2
  JOURNAL   Int. J. Parasitol. 43 (7), 515-520 (2013)
  PUBMED    23500073
REFERENCE   2  (bases 1 to 7405)
  AUTHORS   Massey HC Jr., Ranjit N, Stoltzfus JD, Castelletto ML, Lok JB.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-DEC-2012) Pathobiology, University of Pennsylvania 
            Veterinary School, 3800 Spruce Street, Philadelphia, PA 19104, USA
REFERENCE   3  (bases 1 to 7405)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7405
                     /organism="Expression vector pPV472"
                     /mol_type="other DNA"
                     /PCR_primers="fwd_name: ssd2longpromf, fwd_seq: 
                     ctacataccaatgattatttcaatg, rev_name: ssdaf2promgfpr, 
                     rev_seq: 
                     gaaaagttcttctcctttactcattgatagataattttttttaggcagttaaaaatat 
                     c"
                     /db_xref="taxon:1307126"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        242..260
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      295..2250
                     /regulatory_class="promoter"
                     /note="derived from Strongyloides stercoralis daf-2 
                     promoter region"
     CDS             join(2251..2421,2473..2626,2678..2841,2893..3120)
                     /codon_start=1
                     /product="GFP"
                     /label=GFP
                     /note="green fluorescent protein"
                     /protein_id="AGH70233.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     regulatory      3129..3721
                     /regulatory_class="terminator"
                     /note="derived from Strongyloides stercoralis era-1 
                     terminator"
     protein_bind    complement(3723..3743)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        complement(3778..3796)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3803..3819)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3957..4259
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             4608..5402
                     /codon_start=1
                     /product="Kan/NeoR"
                     /EC_number="2.7.1.95"
                     /label=Kan/NeoR
                     /note="neomycin-kanamycin phosphotransferase type II"
                     /protein_id="AGH70231.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(5652..6512)
                     /codon_start=1
                     /product="AmpR"
                     /EC_number="3.5.2.6"
                     /label=AmpR
                     /note="beta-lactamase"
                     /protein_id="AGH70232.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6636..7224
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"