LOCUS       Exported                7139 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression indicator vector pPV238, complete sequence.
ACCESSION   JX171292
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7139)
  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 7139)
  AUTHORS   Massey HC Jr., Ranjit N, Stoltzfus JD, Castelletto ML, Lok JB.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-JUN-2012) Pathobiology, University of Pennsylvania 
            Veterinary School, 3800 Spruce Street, Philadelphia, PA 19104, USA
REFERENCE   3  (bases 1 to 7139)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7139
                     /organism="Expression indicator vector pPV238"
                     /mol_type="other DNA"
                     /note="Strongyloides stercoralis daf-2 expression 
                     indicator; PCR_primers=rev_name: m13 reverse, rev_seq: 
                     caggaaacagctatgaccatg"
                     /db_xref="taxon:1225555"
     regulatory      7..2766
                     /regulatory_class="promoter"
                     /note="Caenorhabditis elegans daf-2"
     CDS             1401..1415
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             join(2779..2949,3001..3154,3206..3369,3421..3648)
                     /codon_start=1
                     /product="GFP S65C"
                     /label=GFP S65C
                     /note="Aequorea victoria green fluorescent protein"
                     /protein_id="AFQ32906.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFCYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     3UTR           3785..4519
                     /label=Caenorhabditis elegans unc-54 3UTR
                     /note="Caenorhabditis elegans unc-54 3UTR"
     promoter        4884..4988
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4989..5849
                     /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             5187..5849
                     /codon_start=1
                     /product="AmpR"
                     /EC_number="3.5.2.6"
                     /label=AmpR
                     /note="beta-lactamase; ampicillin resistance"
                     /protein_id="AFQ32907.1"
                     /translation="MSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHL
                     TDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEA
                     IPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFI
                     ADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     rep_origin      5944..6626
     rep_origin      6020..6608
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6896..6917
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6932..6962
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6970..6986
                     /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     6994..7014
                     /label=M13 reverse primer
                     /note="M13 reverse primer"
     primer_bind     6994..7010
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"