LOCUS       40924_35663        5567 bp DNA     circular SYN 18-DEC-2018
DEFINITION  PiggyBac_helper_T7_transposase vector pPV257, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5567)
  AUTHORS   Shao H, Li X, Nolan TJ, Massey HC Jr., Pearce EJ, Lok JB.
  TITLE     Transposon-mediated Chromosomal Integration of Transgenes in the 
            Parasitic Nematode Strongyloides ratti and Establishment of Stable 
            Transgenic Lines
  JOURNAL   PLoS Pathog. 8 (8), E1002871 (2012)
  PUBMED    22912584
REFERENCE   2  (bases 1 to 5567)
  AUTHORS   Shao H, Li X, Nolan TJ, Massey HC Jr., Pearce EJ, Lok JB.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-MAY-2012) Pathobiology, University of Pennsylvania, 
            3800 Spruce Street, Philadelphia, PA 19104, USA
REFERENCE   3  (bases 1 to 5567)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5567)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS 
            Pathog."; date: "2012"; volume: "8"; issue: "8"; pages: "E1002871"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (04-MAY-2012) Pathobiology, University of Pennsylvania, 3800 Spruce 
            Street, Philadelphia, PA 19104, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(255..1046)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(1398..1697)
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1835..1851
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1858..1876
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1977..3758
                     /codon_start=1
                     /label=PBase
                     /note="Native piggyBac transposase from Trichoplusia ni"
                     /translation="MGCSLDDEHILSALLQSDDELVGEDSDSEISDHVSEDDVQSDTEE
                     AFIDEVHEVQPTSSGSEILDEQNVIEQPGSSLASNRILTLPQRTIRGKNKHCWSTSKST
                     RRSRVSALNIVRSQRGPTRMCRNIYDPLLCFKLFFTDEIISEIVKWTNAEISLKRRESM
                     TGATFRDTNEDEIYAFFGILVMTAVRKDNHMSTDDLFDRSLSMVYVSVMSRDRFDFLIR
                     CLRMDDKSIRPTLRENDVFTPVRKIWDLFIHQCIQNYTPGAHLTIDEQLLGFRGRCPFR
                     MYIPNKPSKYGIKILMMCDSGTKYMINGMPYLGRGTQTNGVPLGEYYVKELSKPVHGSC
                     RNITCDNWFTSIPLAKNLLQEPYKLTIVGTVRSNKREIPEVLKNSRSRPVGTSMFCFDG
                     PLTLVSYKPKPAKMVYLLSSCDEDASINESTGKPQMVMYYNQTKGGVDTLDQMCSVMTC
                     SRKTNRWPMALLYGMINIACINSFIIYSHNVSSKGEKVQSRKKFMRNLYMSLTSSFMRK
                     RLEAPTLKRYLRDNISNILPNEVPGTSDDSTEEPVMKKRTYCTYCPSKIRRKANASCKK
                     CKKVICREHNIDMCQSCF"
     promoter        complement(4074..4092)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4110..4126)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4134..4150)
                     /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)."
     promoter        complement(4158..4188)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4203..4224)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4512..5100)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5241..5567,1..45))
                     /codon_start=1
                     /label=BleoR
                     /note="antibiotic-binding protein"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
ORIGIN
        1 gtcgcgcgcg gtgagcaccg gaacggcact ggtcaacttg gccatggtgg ccctcctcac
       61 gtgctattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
      121 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgt
      181 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gaaattgtaa
      241 gcgttaataa ttcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg
      301 ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca
      361 gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca
      421 cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg
      481 ccatgggtca cgacgagatc ctcgccgtcg ggcatgctcg ccttgagcct ggcgaacagt
      541 tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct
      601 tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta
      661 gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca
      721 ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc
      781 cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc
      841 cacgatagcc gcgctgcctc gtcttgcagt tcattcaggg caccggacag gtcggtcttg
      901 acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg
      961 attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct
     1021 gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcagag
     1081 cttgatcccc tgcgccatca gatccttggc ggcgagaaag ccatccagtt tactttgcag
     1141 ggcttcccaa ccttaccaga gggcgcccca gctggcaatt ccggttcgct tgctgtccat
     1201 aaaaccgccc agtctagcta tcgccatgta agcccactgc aagctacctg ctttctcttt
     1261 gcgcttgcgt tttcccttgt ccagatagcc cagtagctga cattcatccg gggtcagcac
     1321 cgtttctgcg gactggcttt ctacgtgaaa aggatctagg tgaagatcct ttttgataat
     1381 ctcatgcctg acatttatat tccccagaac atcaggttaa tggcgttttt gatgtcattt
     1441 tcgcggtggc tgagatcagc cacttcttcc ccgataacgg agaccggcac actggccata
     1501 tcggtggtca tcatgcgcca gctttcatcc ccgatatgca ccaccgggta aagttcacgg
     1561 gagactttat ctgacagcag acgtgcactg gccaggggga tcaccatccg tcgccccggc
     1621 gtgtcaataa tatcactctg tacatccaca aacagacgat aacggctctc tcttttatag
     1681 gtgtaaacct taaactgccg tacgtatagg ctgcgcaact gttgggaagg gcgatcggtg
     1741 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     1801 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa
     1861 tacgactcac tatagggcga attgggccct ctagatgcat gctcgagcgg ccgccagtgt
     1921 gatggatatc tgcagaattc gcccttcctg cagcccgggt accggtatat aataaaatgg
     1981 gatgttcttt agacgatgag catatcctct ctgctcttct gcaaagcgat gacgagcttg
     2041 ttggtgagga ttctgacagt gaaatatcag atcacgtaag tgaagatgac gtccagagcg
     2101 atacagaaga agcgtttata gatgaggtac atgaagtgca gccaacgtca agcggtagtg
     2161 aaatattaga cgaacaaaat gttattgaac aaccaggttc ttcattggct tctaacagaa
     2221 tcttgacctt gccacagagg actattagag gtaagaataa acattgttgg tcaacttcaa
     2281 agtccacgag gcgtagccga gtctctgcac tgaacattgt cagatctcaa agaggtccga
     2341 cgcgtatgtg ccgcaatata tatgacccac ttttatgctt caaactattt tttactgatg
     2401 agataatttc ggaaattgta aaatggacaa atgctgagat atcattgaaa cgtcgggaat
     2461 ctatgacagg tgctacattt cgtgacacga atgaagatga aatctatgct ttctttggta
     2521 ttctggtaat gacagcagtg agaaaagata accacatgtc cacagatgac ctctttgatc
     2581 gatctttgtc aatggtgtac gtctctgtaa tgagtcgtga tcgttttgat tttttgatac
     2641 gatgtcttag aatggatgac aaaagtatac ggcccacact tcgagaaaac gatgtattta
     2701 ctcctgttag aaaaatatgg gatctcttta tccatcagtg catacaaaat tacactccag
     2761 gggctcattt gaccatagat gaacagttac ttggttttag aggacggtgt ccgtttagga
     2821 tgtatatccc aaacaagcca agtaagtatg gaataaaaat cctcatgatg tgtgacagtg
     2881 gtacgaagta tatgataaat ggaatgcctt atttgggaag aggaacacag accaacggag
     2941 taccactcgg tgaatactac gtgaaggagt tatcaaagcc tgtgcacggt agttgtcgta
     3001 atattacgtg tgacaattgg ttcacctcaa tccctttggc aaaaaactta ctacaagaac
     3061 cgtataagtt aaccattgtg ggaaccgtgc gatcaaacaa acgcgagata ccggaagtac
     3121 tgaaaaacag tcgctccagg ccagtgggaa catcgatgtt ttgttttgac ggacccctta
     3181 ctctcgtctc atataaaccg aagccagcta agatggtata cttattatca tcttgtgatg
     3241 aggatgcttc tatcaacgaa agtaccggta aaccgcaaat ggttatgtat tataatcaaa
     3301 ctaaaggcgg agtggacacg ctagaccaaa tgtgttctgt gatgacctgc agtaggaaga
     3361 cgaataggtg gcctatggca ttattgtacg gaatgataaa cattgcctgc ataaattctt
     3421 ttattatata cagccataat gtcagtagca agggagaaaa ggttcaaagt cgcaaaaaat
     3481 ttatgagaaa cctttacatg agcctgacgt catcgtttat gcgtaagcgt ttagaagctc
     3541 ctactttgaa gagatatttg cgcgataata tctctaatat tttgccaaat gaagtgcctg
     3601 gtacatcaga tgacagtact gaagagccag taatgaaaaa acgtacttac tgtacttact
     3661 gcccctctaa aataaggcga aaggcaaatg catcgtgcaa aaaatgcaaa aaagttattt
     3721 gtcgagagca taatattgat atgtgccaaa gttgtttctg actgactaat aagtataatt
     3781 tgtttctatt atgtataagt taagctaatt acttatttta taatacaaca tgactgtttt
     3841 taaagtacaa aataagttta tttttgtaaa agagagaatg tttaaaagtt ttgttacttt
     3901 agaagaaatt ttgagttttt gttttttttt aataaataaa taaacataaa ttgtttgttg
     3961 aatttggatc ccctaggaat tcagcggccg ccacaagggc gaattccagc acactggcgg
     4021 ccgttactag tggatccgag ctcggtacca agcttgatgc atagcttgag tattctatag
     4081 tgtcacctaa atagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4141 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4201 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4261 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4321 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4381 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4441 gcaggaaaga acatgtgagc aaaaggccag caaaagccca ggaaccgtaa aaaggccgcg
     4501 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4561 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4621 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4681 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4741 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4801 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4861 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4921 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     4981 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5041 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5101 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5161 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5221 tgaagtttta gcacgtgtca gtcctgctcc tcggccacga agtgcacgca gttgccggcc
     5281 gggtcgcgca gggcgaactc ccgcccccac ggctgctcgc cgatctcggt catggccggc
     5341 ccggaggcgt cccggaagtt cgtggacacg acctccgacc actcggcgta cagctcgtcc
     5401 aggccgcgca cccacaccca ggccagggtg ttgtccggca ccacctggtc ctggaccgcg
     5461 ctgatgaaca gggtcacgtc gtcccggacc acaccggcga agtcgtcctc cacgaagtcc
     5521 cgggagaacc cgagccggtc ggtccagaac tcgaccgctc cggcgac
//
