LOCUS       62056_1216        9633 bp DNA     circular SYN 14-MAR-2021
DEFINITION  Cloning vector Hr5/Ie1-tTAV(OX5378), complete sequence.
ACCESSION   MT533615
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9633)
  AUTHORS   Harvey-Samuel TD, Xu X, Lovett E, Dafa'alla T, Walker A, Norman VC, 
            Carter R, Teal J, Akilan L, Leftwich PT, Reitmayer CM, Siddiqui HA, 
            Alphey L.
  TITLE     Engineered expression of the invertebrate-specific scorpion toxin 
            AaHIT reduces adult longevity and female fecundity in the 
            diamondback moth Plutella xylostella
  JOURNAL   Pest Manag Sci (2021) In press
  PUBMED    33660916
REFERENCE   2  (bases 1 to 9633)
  AUTHORS   Harvey-Samuel TD.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-MAY-2020) Arthropod Genetics, The Pirbright Institute,
            Ash Road, Woking, Surrey GU24 0NF, United Kingdom
REFERENCE   3  (bases 1 to 9633)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Pest Manag 
            Sci (2021) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (27-MAY-2020) Arthropod Genetics, The Pirbright Institute, Ash Road,
            Woking, Surrey GU24 0NF, United Kingdom"
FEATURES             Location/Qualifiers
     source          1..9633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  8..673
                     /mobile_element_type="transposon:Piggybac"
                     /label=bac
     enhancer        744..1226
                     /label=hr5 enhancer
                     /note="baculovirus early transcription enhancer"
     CDS             2025..2645
                     /label=TetR
                     /note="tetracycline repressor TetR"
     CDS             2802..3035
                     /label=VP16 AD
                     /note="transcriptional activation domain of herpes simplex
                     virus protein VP16 (Triezenberg et al., 1988; Cousens et 
                     al., 1989)"
     regulatory      3487..4027
                     /regulatory_class="promoter"
     CDS             4049..4741
                     /label=ZsGreen1
                     /note="Zoanthus green fluorescent protein"
     CDS             4760..4780
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     polyA_signal    4921..5042
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     mobile_element  5087..6133
                     /mobile_element_type="transposon:Piggybac"
                     /label=bac
     primer_bind     complement(6836..6852)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7326..7430
                     /label=AmpR promoter
     CDS             7431..8288
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      8462..9050
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    9338..9359
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        9374..9404
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9412..9428
                     /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     9436..9452
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tttttaaccc tagaaagata gtctgcgtaa aattgacgca tgcattcttg aaatattgct
       61 ctctctttct aaatagcgcg aatccgtcgc tgtgcattta ggacatctca gtcgccgctt
      121 ggagctcccg tgaggcgtgc ttgtcaatgc ggtaagtgtc actgattttg aactataacg
      181 accgcgtgag tcaaaatgac gcatgattat cttttacgtg acttttaaga tttaactcat
      241 acgataatta tattgttatt tcatgttcta cttacgtgat aacttattat atatatattt
      301 tcttgttata gatatcgtga ctaatatata ataaaatggg tagttcttta gacgatgagc
      361 atatcctctc tgctcttctg caaagcgatg acgagcttgt tggtgaggat tctgacagtg
      421 aaatatcaga tcacgtaagt gaagatgacg tccagagcga tacagaagaa gcgtttatag
      481 atgaggtaca tgaagtgcag ccaacgtcaa gcggtagtga aatattagac gaacaaaatg
      541 ttattgaaca accaggttct tcattggctt ctaacagaat cttgaccttg ccacagagga
      601 ctattagagg taagaataaa cattgttggt caacttcaaa gtccacgagg cgtagccgag
      661 tctctgcact gaacattgtc agatcggccc gggatgatgt catttgtttt taaaattgaa
      721 ctggctttac gagtagaatt ctacgcgtaa aacacaatca agtatgagtc ataagctgat
      781 gtcatgtttt gcacacggct cataaccgaa ctggctttac gagtagaatt ctacttgtaa
      841 cgcacgatca gtggatgatg tcatttgttt ttcaaatcga gatgatgtca tgttttgcac
      901 acggctcata aactcgcttt acgagtagaa ttctacgtgt aacgcacgat cgattgatga
      961 gtcatttgtt ttgcaatatg atatcataca atatgactca tttgtttttc aaaaccgaac
     1021 ttgatttacg ggtagaattc tacttgtaaa gcacaatcaa aaagatgatg tcatttgttt
     1081 ttcaaaactg aactcgcttt acgagtagaa ttctacgtgt aaaacacaat caagaaatga
     1141 tgtcatttgt tataaaaata aaagctgatg tcatgttttg cacatggctc ataactaaac
     1201 tcgctttacg ggtagaattc tacgcgtaaa acatgattga taattaaata attcatttgc
     1261 aagctatacg ttaaatcaaa cggacgctcg aggttgcaca acactattat cgatttgcag
     1321 ttcgggacat aaatgtttaa atatatcgat gtctttgtga tgcgcgcgac atttttgtag
     1381 gttattgata aaatgaacgg atacgttgcc cgacattatc attaaatcct tggcgtagaa
     1441 tttgtcgggt ccattgtccg tgtgcgctag catgcccgta acggacctcg tacttttggc
     1501 ttcaaaggtt ttgcgcacag acaaaatgtg ccacacttgc agctctgcat gtgtgcgcgt
     1561 taccacaaat cccaacggcg cagtgtactt gttgtatgca aataaatctc gataaaggcg
     1621 cggcgcgcga atgcagctga tcacgtacgc tcctcgtgtt ccgttcaagg acggtgttac
     1681 cgacctcaga ttaatgttta tcggccgact gttttcgtat ccgctcacca aacgcgtttt
     1741 tgcattaaca ttgtatgtcg gcggatgttc tatatctaat ttgaataaat aaacgataac
     1801 cgcgttggtt ttagagggca taataaaaga aatattgtta tcgtgttcgc cattagggca
     1861 gtataaattg acgttcatgt tggatattgt ttcagttgca agttgacact ggcggcgaca
     1921 agacaattct aattggggta agttttcccg ttcttttctg ggttcttccc ttttgctcat
     1981 ccttgctgca ctaccttcag gtgcaagttg agattcaggc caccatgagc cgcctggata
     2041 agtccaaagt catcaactcc gcgttggagc tgttgaacga agttggcatt gagggactga
     2101 cgacccgcaa gttggcgcag aagctgggcg tggagcagcc caccctctac tggcacgtga
     2161 agaataagcg ggcgctgctg gatgccctgg ccatcgagat gctcgaccgc caccacacgc
     2221 atttttgccc gttggaaggc gagtcctggc aggacttcct ccgcaataac gccaagtcgt
     2281 tccgctgcgc tctgctgtcc caccgagacg gtgccaaagt ccatctcggc acgcgcccga
     2341 ccgaaaagca atacgagaca ctggagaacc agctcgcgtt cctgtgccag caaggcttca
     2401 gcctggaaaa tgctctctac gctctgagcg ccgtcggtca ctttaccctg ggctgcgtgc
     2461 tggaggacca agagcatcaa gtcgcaaaag aggagcgcga gaccccaaca accgattcga
     2521 tgcccccact gctgcgtcag gcaatcgagc tgttcgatca tcaaggagcc gagccggcat
     2581 tcctgttcgg cttggagctg attatctgcg gattggaaaa gcaactgaaa tgcgagtcgg
     2641 gctcgggccc cgcgtacagc cgcgcgcgta cgaaaaacaa ttacgggtct accatcgagg
     2701 gcctgctcga tctcccggac gacgacgccc ccgaagaggc ggggctggcg gctccgcgcc
     2761 tgtcctttct ccccgcggga cacacgcgca gactgtcgac ggcccccccg accgatgtca
     2821 gcctggggga cgagctccac ttagacggcg aggacgtggc gatggcgcat gccgacgcgc
     2881 tagacgattt cgatctggac atgttggggg acggggattc cccgggtccg ggatttaccc
     2941 cccacgactc cgccccctac ggcgctctgg atatggccga cttcgagttt gagcagatgt
     3001 ttaccgatgc ccttgggatt gacgagtacg gtgggtaagg atccagccaa atagaaaatt
     3061 attcagttcc tggcttaagt ttttaaaagt gatattattt atttggttgt aaccaaccaa
     3121 aagaatgtaa ataactaata cataattatg ttagttttaa gttagcaaca aattgatttt
     3181 agctatatta gctacttggt taataaatag aatatattta tttaaagata attgcgtttt
     3241 tattgtcagg gagtgagttt gcttaaaaac tcgtttagat ctgtcctcga gaaattattt
     3301 atttaaatgc gatggagagt tggcgccgaa tcgaaaactt tacgcgctta aaagcacgag
     3361 ttggcatccc tagtaaacag ctgttcgtga agatatgcag tgcaaacgaa aaacccgcct
     3421 acaaacattg ttattttgat tagattacgg attacaaaat ggaaccggcg cgcggccgca
     3481 tggtaccata ctcggtggcc tccccaccac caactttttt gcactgcaaa aaaacacgct
     3541 tttgcacgcg ggcccataca tagtacaaac tctacgtttc gtagactatt ttacataaat
     3601 agtctacacc gttgtatacg ctccaaatac actaccacac attgaacctt tttgcagtgc
     3661 aaaaaagtac gtgtcggcag tcacgtaggc cggccttatc gggtcgcgtc ctgtcacgta
     3721 cgaatcacat tatcggaccg gacgagtgtt gtcttatcgt gacaggacgc cagcttcctg
     3781 tgttgctaac cgcagccgga cgcaactcct tatcggaaca ggacgcgcct ccatatcagc
     3841 cgcgcgttat ctcatgcgcg tgaccggaca cgaggcgccc gtcccgctta tcgcgcctat
     3901 aaatacagcc cgcaacgatc tggtaaacac agttgaacag catctgttac agcgacacaa
     3961 catgagccgg tccaacaacg ccaacgcgcc cacgccatcc aaccgccgcc gcaacctgtc
     4021 tctggtggat ccgctaccgg tcgccaccat ggcccagtcc aagcacggcc tgaccaagga
     4081 gatgaccatg aagtaccgca tggagggctg cgtggacggc cacaagttcg tgatcaccgg
     4141 cgagggcatc ggctacccct tcaagggcaa gcaggccatc aacctgtgcg tggtggaggg
     4201 cggccccttg cccttcgccg aggacatctt gtccgccgcc ttcatgtacg gcaaccgcgt
     4261 gttcaccgag tacccccagg acatcgtcga ctacttcaag aactcctgcc ccgccggcta
     4321 cacctgggac cgctccttcc tgttcgagga cggcgccgtg tgcatctgca acgccgacat
     4381 caccgtgagc gtggaggaga actgcatgta ccacgagtcc aagttctacg gcgtgaactt
     4441 ccccgccgac ggccccgtga tgaagaagat gaccgacaac tgggagccct cctgcgagaa
     4501 gatcatcccc gtgcccaagc agggcatctt gaagggcgac gtgagcatgt acctgctgct
     4561 gaaggacggt ggccgcttgc gctgccagtt cgacaccgtg tacaaggcca agtccgtgcc
     4621 ccgcaagatg cccgactggc acttcatcca gcacaagctg acccgcgagg accgcagcga
     4681 cgccaagaac cagaagtggc acctgaccga gcacgccatc gcctccggct ccgccttgcc
     4741 ctccggactc agatctcgac ccaagaaaaa gcggaaggtg gaggacccgt aagatccacc
     4801 ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta
     4861 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     4921 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4981 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     5041 taacgcgagc ctagcggccg ctcatttaaa tctggccggc cgcaaccatt gtgggaaccg
     5101 tgcgatcaaa caaacgcgag ataccggaag tactgaaaaa cagtcgctcc aggccagtgg
     5161 gaacatcgat gttttgtttt gacggacccc ttactctcgt ctcatataaa ccgaagccag
     5221 ctaagatggt atacttatta tcatcttgtg atgaggatgc ttctatcaac gaaagtaccg
     5281 gtaaaccgca aatggttatg tattataatc aaactaaagg cggagtggac acgctagacc
     5341 aaatgtgttc tgtgatgacc tgcagtagga agacgaatag gtggcctatg gcattattgt
     5401 acggaatgat aaacattgcc tgcataaatt cttttattat atacagccat aatgtcagta
     5461 gcaagggaga aaaggtccaa agtcgcaaaa aatttatgag aaacctttac atgagcctga
     5521 cgtcatcgtt tatgcgtaag cgtttagaag ctcctacttt gaagagatat ttgcgcgata
     5581 atatctctaa tattttgcca aatgaagtgc ctggtacatc agatgacagt actgaagagc
     5641 cagtaatgaa aaaacgtact tactgtactt actgcccctc taaaataagg cgaaaggcaa
     5701 atgcatcgtg caaaaaatgc aaaaaagtta tttgtcgaga gcataatatt gatatgtgcc
     5761 aaagttgttt ctgactgact aataagtata atttgtttct attatgtata agttaagcta
     5821 attacttatt ttataataca acatgactgt ttttaaagta caaaataagt ttatttttgt
     5881 aaaagagaga atgtttaaaa gttttgttac tttatagaag aaattttgag tttttgtttt
     5941 tttttaataa ataaataaac ataaataaat tgtttgttga atttattatt agtatgtaag
     6001 tgtaaatata ataaaactta atatctattc aaattaataa ataaacctcg atatacagac
     6061 cgataaaaca catgcgtcaa ttttacgcat gattatcttt aacgtacgtc acaatatgat
     6121 tatctttcta gggttaaata atagtttcta atttttttat tattcagcct gctgtcgtga
     6181 ataccgtata tctcaacgct gtctgtgaga ttgtcgtatt ctagcctttt tagtttttcg
     6241 ctcatcgact tgatattgtc cgacacattt tcgtcgattt gcgttttgat caaagacttg
     6301 agcagagaca cgttaatcaa ctgttcaaat tgatccatat taacgatatc aacccgatgc
     6361 gtatatggtg cgtaaaatat attttttaac cctcttatac tttgcactct gcgttaatac
     6421 gcgttcgtgt acagacgtaa tcatgttttc ttttttggat aaaactccta ctgagtttga
     6481 cctcatatta gaccctcaca agttgcaaaa cgtggcattt tttaccaatg aagaatttaa
     6541 agttatttta aaaaatttca tcacagattt aaagaagaac caaaaattaa attatttcaa
     6601 cagtttaatc gaccagttaa tcaacgtgta cacagacgcg tcggcaaaaa acacgcagcc
     6661 cgacgtgttg gctaaaatta ttaaatcaac ttgtgttata gtcacggatt tgccgtccaa
     6721 cgtgttcctc aaaaagttga agaccaacaa gtttacggac actattaatt atttgatttt
     6781 gccccacttc attttgtggg atcacaattt tgttatattt taaacaaagc ttggcactgg
     6841 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     6901 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     6961 cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc
     7021 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg
     7081 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     7141 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     7201 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     7261 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     7321 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     7381 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     7441 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     7501 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     7561 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     7621 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     7681 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     7741 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     7801 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     7861 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     7921 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     7981 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     8041 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     8101 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     8161 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     8221 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     8281 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     8341 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     8401 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     8461 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     8521 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     8581 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     8641 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     8701 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     8761 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     8821 cctacaccga actgagatac ctacagcgtg agcattgaga aagcgccacg cttcccgaag
     8881 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     8941 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     9001 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     9061 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     9121 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     9181 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     9241 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     9301 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     9361 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     9421 gataacaatt tcacacagga aacagctatg accatgatta cgaatttcga cctgcaggca
     9481 tgcaagcttg catgcctgca ggtcgacgct cgcgcgactt ggtttgccat tctttagcgc
     9541 gcgtcgcgtc acacagcttg gccacaatgt ggtttttgtc aaacgaagat tctatgacgt
     9601 gtttaaagtt taggtcgagt aaagcgcaaa tct
//
