LOCUS       40924_35658        6758 bp DNA     circular SYN 18-DEC-2018
DEFINITION  PiggyBac_based_Ss_Act2_gfp_era1 vector pPV254, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6758)
  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 6758)
  AUTHORS   Shao H, Li X, Nolan TJ, Massey HC Jr., Pearce EJ, Lok JB.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-MAY-2012) Pathobiology, University of Pennsylvania, 
            3800 Spruce Street, Philadelphia, PA 19104, USA
REFERENCE   3  (bases 1 to 6758)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6758)
  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 
            (03-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..6758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..128
                     /label=AmpR promoter
     CDS             129..986
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1160..1748
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /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
                     /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"
     mobile_element  2641..2675
                     /mobile_element_type="transposon:piggyBac ITR"
                     /label=ITR
     CDS             complement(3315..3326)
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
     misc_recomb     3826..3830
                     /gene="act2"
                     /label=target sequence
                     /note="target sequence"
     CDS             join(4249..4419,4471..4624,4676..4839,4891..5118)
                     /codon_start=1
                     /product="GFP"
                     /label=GFP
                     /protein_id="AFN89784.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFCYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     3'UTR           5138..5724
                     /gene="era1"
     gene            5138..5724
                     /gene="era1"
                     /label=era1
                     /note="Ss-era1"
     primer_bind     5738..5754
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     mobile_element  5951..6013
                     /mobile_element_type="transposon:piggyBac ITR"
                     /label=ITR
     misc_recomb     6014..6017
                     /label=target sequence
                     /note="target sequence"
     primer_bind     complement(6143..6159)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6301..6756
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
       61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
      841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
      901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
      961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     1921 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     1981 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     2041 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     2101 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     2161 ccaagcgcgc ccgccgggta actcacgggg tatccatgtc catttctgcg gcatccagcc
     2221 aggatacccg tcctcgctga cgtaatatcc cagcgccgca ccgctgtcat taatctgcac
     2281 accggcacgg cagttccggc tgtcgccggt attgttcggg ttgctgatgc gcttcgggct
     2341 gaccatccgg aactgtgtcc ggaaaagccg cgacgaactg gtatcccagg tggcctgaac
     2401 gaacagttca ccgttaaagg cgtgcatggc cacaccttcc cgaatcatca tggtaaacgt
     2461 gcgttttcgc tcaacgtcaa tgcagcagca gtcatcctcg gcaaactctt tccatgccgc
     2521 ttcaacctcg cgggaaaagg cacgggcttc ttcctccccg atgcccagat agcgccagct
     2581 tgggcgatga ctgagccgga aaaaagaccc gacgatatga tcctgatgca gctagattaa
     2641 ccctagaaag atagtctgcg taaaattgac gcatgcattc ttgaaatatt gctctctctt
     2701 tctaaatagc gcgaatccgt cgctgtgcat ttaggacatc tcagtcgccg cttggagctc
     2761 ccgtgaggcg tgcttgtcaa tgcggtaagt gtcactgatt ttgaactata acgaccgcgt
     2821 gagtcaaaat gacgcatgat tatcttttac gtgactttta agatttaact catacgataa
     2881 ttatattgtt atttcatgtt ctacttacgt gataacttat tatatatata ttttcttgtt
     2941 atagatatca agcttgatta tcgtagtgag aggacgtgta gagataataa atttcccaac
     3001 ttaatctatt ataggaactt agttaaaaat gattgtattg gtcctctttg tgaaagtgtg
     3061 gaaaagaggt acctcgtagt ttttttcttc tcaattctca tcatcaccct gtgtaacatc
     3121 acatctcata tagagaaatc gcgactttta ttgcttagca tatactttgt aagttgtttt
     3181 cactgaggct ttgtccatca cacgctacac attatcgctg ttatttttat tatttatatt
     3241 tattagtaat aatattaaat aaacacatag gaattttgtc tttgttggcc cttccagcct
     3301 aaatagttat aaaatctacc ttctattata catacgcata tgttatttaa aaagttgaag
     3361 tgatttctct tctacagaaa ctacacattt atcttcttat tttcttccaa acatatagca
     3421 ataataattt aaaacttttt cttactcctc ccagagtttg ttaagttgat catatttaaa
     3481 attcttctgt cttctacaac aaataatata tttattataa caccaactac ctcgttccaa
     3541 gtattacgta ggagcatata cataccacaa tgcttaggaa atgggggcgg ttatgatata
     3601 acattatttc aatgtaacat tcttttttac cacatatatt ttctactgaa ttctgaaaac
     3661 tcaatttcaa acctcctata tatatatatt gttaggcact tctaagatta taattcattt
     3721 aactctctta aatttatcta ttttaaattt ttcctttttt gtattattta tactattata
     3781 tcgaaacttt attattacat gtactattta acttttattt tcttatataa taataatact
     3841 aaataataac tgataatcgt atctgtagag attattgaat tgtgattagt aatgtaaagt
     3901 aaatctatat taagatatct caaagatgca ctcaagttat ttgaaaaatt tcttatacat
     3961 agattaatac ttatattaag aaactttaaa aaaaagtttt aaataaaaaa aaaaattcac
     4021 aattgtacta ttttatcttg tcgttcagtt tatcaccgta gtatttttat ggctagtaat
     4081 attcatcatt tttagataaa ctaacttgct atagaaaatg tgtgacgaag aggttgccgc
     4141 tgcaggtcga ctagaggatc cccgggattg gccaaaggac ccaaaggtat gtttcgaatg
     4201 atactaacat aacatagaac attttcagga ggacccttgg agggtaccat gagtaaagga
     4261 gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg
     4321 cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt
     4381 aaatttattt gcactactgg aaaactacct gttccatggg taagtttaaa catatatata
     4441 ctaactaacc ctgattattt aaattttcag ccaacacttg tcactacttt ctgttatggt
     4501 gttcaatgct tctcgagata cccagatcat atgaaacggc atgacttttt caagagtgcc
     4561 atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag
     4621 acacgtaagt ttaaacagtt cggtactaac taaccataca tatttaaatt ttcaggtgct
     4681 gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg agttaaaagg tattgatttt
     4741 aaagaagatg gaaacattct tggacacaaa ttggaataca actataactc acacaatgta
     4801 tacatcatgg cagacaaaca aaagaatgga atcaaagttg taagtttaaa catgatttta
     4861 ctaactaact aatctgattt aaattttcag aacttcaaaa ttagacacaa cattgaagat
     4921 ggaagcgttc aactagcaga ccattatcaa caaaatactc caattggcga tggccctgtc
     4981 cttttaccag acaaccatta cctgtccaca caatctgccc tttcgaaaga tcccaacgaa
     5041 aagagagacc acatggtcct tcttgagttt gtaacagctg ctgggattac acatggcatg
     5101 gatgaactat acaaatagca ttcgtagaat tcctaggaat ttattttgat aagtctcgcg
     5161 tattttattt gtaataatct ttttataaga tcaatttaca tttatactga taaaattgac
     5221 acaattacac tttttattat aaaatatttt aaaatatttc tatcaaaagg tttatttttg
     5281 ttttaaatat aatggttttt tttgtaaaaa atacttattt cgtttgataa tctttattca
     5341 gagatttgaa taattttaaa taaagtagtt tagttttcat ataaaagtta atggtatata
     5401 taaatattta aatagtaaag attgaatcag tattcaaaat gacacctaaa agttattcta
     5461 tttctagcta taacaataat aataaagata attatgatac agatttagaa aaaaaaataa
     5521 ggttaaatta attaattcta tttttaatta aaatgattta agggagttaa atgaaacaga
     5581 aacagattgg gtctctatat atttagttta tgtaatgtta ttttttggtg gaatacaaat
     5641 aacggtatat tttacttcaa tgtggccata cttatcaaca ataaatagta aggcaaatgt
     5701 agatacttta gggtgggttg tagcactagt cggccgctct agaactagtg gatcccccgg
     5761 gctgcaggaa ttcgataaaa gttttgttac tttatagaag aaattttgag tttttgtttt
     5821 tttttaataa ataaataaac ataaataaat tgtttgttga atttattatt agtatgtaag
     5881 tgtaaatata ataaaactta atatctattc aaattaataa ataaacctcg atatacagac
     5941 cgataaaaca catgcgtcaa ttttacgcat gattatcttt aacgtacgtc acaatatgat
     6001 tatctttcta gggttaatct agctgcgtgt tctgcagcgt gtcgagcatc ttcatctgct
     6061 ccatcacgct gtaaaacaca tttgcaccgc gagtctgccc gtcctccacg ggttcaaaaa
     6121 cgtgaatgaa cgaggcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     6181 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     6241 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg
     6301 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     6361 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     6421 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     6481 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     6541 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     6601 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     6661 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     6721 acgcgaattt taacaaaata ttaacgctta caatttag
//
