LOCUS       Exported                8024 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPV512, complete sequence.
ACCESSION   KU665997
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8024)
  AUTHORS   Lok JB, Shao H, Massey HC, Li X.
  TITLE     Transgenesis in Strongyloides and related parasitic nematodes: 
            historical perspectives, current functional genomic applications and
            progress towards gene disruption and editing
  JOURNAL   Parasitology (2016) In press
  PUBMED    27000743
REFERENCE   2  (bases 1 to 8024)
  AUTHORS   Lok JB, Shao H, Massey HC Jr., Li X.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2016) Pathobiology, University of Pennsylvania, 
            3800 Spruce Street, Philadelphia, PA 19104, USA
REFERENCE   3  (bases 1 to 8024)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8024
                     /organism="Cloning vector pPV512"
                     /mol_type="other DNA"
                     /db_xref="taxon:1821230"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             129..989
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="Amp-r"
                     /protein_id="ANB62409.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2072..2102
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /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     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      2949..4150
                     /regulatory_class="promoter"
                     /note="Act2; promoter"
     CDS             complement(3315..3326)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    4259..5124
                     /label=similar to green fluorescent protein
                     /note="similar to green fluorescent protein"
     misc_feature    5131..5628
                     /label=DHFR
                     /note="DHFR"
     CDS             5132..5599
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="SLIAALAVDHVIGMETVMPWNLPADLAWFKRNTLNKPVIMGRHTW
                     ESIGRPLPGRKNIILSSQPSTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFLPK
                     AQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILER"
     CDS             5603..5629
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    5629..5716
                     /note="P2A; linker"
     CDS             5648..5704
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             5717..6415
                     /codon_start=1
                     /product="red fluorescent protein"
                     /label=red fluorescent protein
                     /note="RFP"
                     /protein_id="ANB62410.1"
                     /translation="MASSEDVIKEFMRFKVKMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERIMNFEDGGV
                     VTVTQDSTLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYKAKKQVQLPGAYKTDIKLDIISHNEDYTIVEQYERAEGRHS
                     TGAGSPRNLF"
     misc_feature    6428..6990
                     /note="Era1; UTR"
     primer_bind     7004..7020
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7409..7425)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      7567..8022
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"