LOCUS       40924_35668        7405 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pPV472, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7405)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Int. J. 
            Parasitol."; date: "2013"; volume: "43"; issue: "7"; pages: 
            "515-520"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (11-DEC-2012) Pathobiology, University of Pennsylvania Veterinary 
            School, 3800 Spruce Street, Philadelphia, PA 19104, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7405
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /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
                     /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
                     /note="derived from Strongyloides stercoralis daf-2
                     promoter region"
                     /regulatory_class="promoter"
     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
                     /note="derived from Strongyloides stercoralis era-1
                     terminator"
                     /regulatory_class="terminator"
     protein_bind    complement(3723..3743)
                     /label=attB3
                     /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..4256
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     CDS             4608..5399
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             complement(5655..6512)
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      6636..7224
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctca
      241 gaattaaccc tcactaaagg gactagtcct gcaggtttaa acgaattcgc ccttctacat
      301 accaatgatt atttcaatga aatttaatta aaacttacaa tttttattgc tacattttaa
      361 atttcaacat aacttttaac atatgttaat attttaaaat taaaaagaca actttaattt
      421 tatgaattat tactggtttt aaatgtggac aaaattttaa aatttaataa aataaaaaac
      481 taactttatt aattgttttt ttgtaataag attcatcaaa tgttatatag aaaatgtgtt
      541 gtattaatga aaaaattaaa atttcatttt taatatatgt tatttattaa aattttaata
      601 gataattttt ttttgtgaaa gacaattatt ataaaaataa aattgcatag aaaatttttt
      661 atgcttattt tttaaataca tttttaatca gaatcaacac ttttatcatt tatcgggtta
      721 tccatttcca tcataatcga atctatcatt aacattggtt ttattattaa tatttccaac
      781 cagcaaggtg aatctgtaat tgattttctt ctataatttt ctccccatcc ttttataaag
      841 cttatacgag tttgataata ttgttgtaga tttattaatc ccattatacc ttctgccctt
      901 tttaacattg tactaaaaac ttcattatca tatatacata tttcagatcc taaaaatttg
      961 aaatttaatt aaaaaaattt ctatccttac ccggagataa acgataaaca catgctggat
     1021 ctgtcaccag atattgcacc ttgcattgga cattgaataa atattggaga tttacattta
     1081 gattgtattt ttatattatc acctattttt attatatgac aaccccctcc aatttgacat
     1141 aatacttttg atatagcaac atttctttct ggagacgtta gtgaacctaa tgaaaatcta
     1201 ttttttgccc catctttatt acaaaatcca tcaactataa ttgtatcact tcttccacga
     1261 aatgtagctc ctgttcttga atgaaattcc caatatttta aatgacacca tgcattatct
     1321 tcataatatg gtaattcact acgatttact gttcttacat acatattttt ttcaatatca
     1381 tcacttgaag gtacaactgt tttattttga ggaaatattg gattaatata ttcttcgttt
     1441 ataatattat cattagtagt atatattact tgattaggtg ttggatatga tgaatgaaaa
     1501 agtacatcat tttgttgttg ttgtacagat gggacaccat atgattgaac atacgtatat
     1561 tgttgaggat atgtatttat ttgagaataa ttactataat ttggaatcat tggttgagaa
     1621 atttgctgct gttggtgttg ttggggataa ccttgattaa tattttgaat ttcattatta
     1681 tcatttaaat tattcatcta aaaaataata aataaaaata gacattgtta ataaaaaaat
     1741 atattaaaac aatttcaacg ctatttttag atttaatacc aaaaaaatat attttttttt
     1801 attatcttta tccctatatt taaaataaat acaacaatct tgtaatgaat tatggaaaaa
     1861 tgtgacaaaa caatgaagta tcatctttcc aaaaatacaa aatagctgct tatatgttgt
     1921 agtttgttct agtattaaca taaagtgaaa atattatttt gtttatctct aataattttt
     1981 taaattaatt cagtctaaac gtcataaatg tatatttatt atcatatata ttaaggataa
     2041 tatcttgctg gcttccctta tcacttggaa attaaaaagt tttcttttaa aaatttataa
     2101 tatctcttcc gatacctttt ttttttattc ttcttacaat tttatttaaa aaagatatat
     2161 ataataaata tacattaaat ctataaaata taagcattat tttagtaata ctaatgatat
     2221 ttttaactgc ctaaaaaaaa ttatctatca atgagtaaag gagaagaact tttcactgga
     2281 gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt ttctgtcagt
     2341 ggagagggtg aaggtgatgc aacatacgga aaacttaccc ttaaatttat ttgcactact
     2401 ggaaaactac ctgttccatg ggtaagttta aacatatata tactaactaa ccctgattat
     2461 ttaaattttc agccaacact tgtcactact ctcacttatg gtgttcaatg cttctcgaga
     2521 tacccagatc atatgaaaca gcatgacttt ttcaagagtg ccatgcccga aggttatgta
     2581 caggaaagaa ctatattttt caaagatgac gggaactaca agacacgtaa gtttaaacag
     2641 ttcggtacta actaaccata catatttaaa ttttcaggtg ctgaagtcaa gtttgaaggt
     2701 gatacccttg ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt
     2761 cttggacaca aattggaata caactataac tcacacaatg tatacatcat ggcagacaaa
     2821 caaaagaatg gaatcaaagt tgtaagttta aacatgattt tactaactaa ctaatctgat
     2881 ttaaattttc agaacttcaa aattagacac aacattgaag atggaagcgt tcaactagca
     2941 gaccattatc aacaaaatac tccaattggc gatggccctg tccttttacc agacaaccat
     3001 tacctgtcca cacaatctgc cctttcgaaa gatcccaacg aaaagagaga ccacatggtc
     3061 cttcttgagt ttgtaacagc tgctgggatt acacatggca tggacgaact atacaaatga
     3121 taactaggaa tttattttga taagtctcgc gtattttatt tgtaataatc tttttataag
     3181 atcaatttac atttatactg ataaaattga cacaattaca ctttttatta taaaatattt
     3241 taaaatattt ctatcaaaag gtttattttt gttttaaata taatggtttt ttttgtaaaa
     3301 aatacttatt tcgtttgata atctttattc agagatttga ataattttaa ataaagtagt
     3361 ttagttttca tataaaagtt aatggtatat ataaatattt aaatagtaaa gattgaatca
     3421 gtattcaaaa tgacacctaa aagttattct atttctagct ataacaataa taataaagat
     3481 aattatgata cagatttaga aaaaaaaata aggttaaatt aattaattct atttttaatt
     3541 aaaatgattt aagggagtta aatgaaacag aaacagattg ggtctctata tatttagttt
     3601 atgtaatgtt attttttggt ggaatacaaa taacggtata ttttacttca atgtggccat
     3661 acttatcaac aataaatagt aaggcaaatg tagatacttt agggtgggtt gtagcactag
     3721 tgcaacttta ttatacaaag ttgaagggcg aattcgcggc cgctaaattc aattcgccct
     3781 atagtgagtc gtattacaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     3841 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     3901 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctatac gtacggcagt
     3961 ttaaggttta cacctataaa agagagagcc gttatcgtct gtttgtggat gtacagagtg
     4021 atattattga cacgccgggg cgacggatgg tgatccccct ggccagtgca cgtctgctgt
     4081 cagataaagt ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca
     4141 tgatgaccac cgatatggcc agtgtgccgg tctccgttat cggggaagaa gtggctgatc
     4201 tcagccaccg cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataaa
     4261 tgtcaggcat gagattatca aaaaggatct tcacctagat ccttttcacg tagaaagcca
     4321 gtccgcagaa acggtgctga ccccggatga atgtcagcta ctgggctatc tggacaaggg
     4381 aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg cgatagctag
     4441 actgggcggt tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggta
     4501 aggttgggaa gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc
     4561 gcaggggatc aagctctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     4621 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4681 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4741 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     4801 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4861 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4921 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4981 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     5041 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     5101 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     5161 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     5221 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     5281 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     5341 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     5401 gaattattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta
     5461 tttcacaccg catcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5521 tttttctaaa tacattcaaa tatgtatccg ctcatgagat tatcaaaaag gatcttcacc
     5581 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5641 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5701 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5761 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5821 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5881 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5941 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6001 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6061 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6121 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6181 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6241 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6301 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6361 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6421 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6481 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6541 atttatcagg gttattgtct catgaccaaa atcccttaac gtgagttttc gttccactga
     6601 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6661 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6721 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6781 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6841 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6901 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6961 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     7021 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7081 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     7141 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     7201 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7261 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     7321 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     7381 gagtcagtga gcgaggaagc ggaag
//
