LOCUS       62056_23420        8763 bp DNA     circular SYN 14-MAR-2021
DEFINITION  Cloning vector tetO-AaHIT(AGG1074), complete sequence.
ACCESSION   MT533613
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8763)
  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 8763)
  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 8763)
  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..8763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     sig_peptide     208..267
                     /label=GP64 signal sequence
                     /note="signal sequence from the baculovirus envelope 
                     glycoprotein GP64"
     mobile_element  698..1370
                     /mobile_element_type="transposon:Piggybac"
                     /label=bac
     primer_bind     complement(1523..1539)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1547..1563)
                     /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(1571..1601)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1616..1637)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1925..2513)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2687..3544)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(3545..3649)
                     /label=AmpR promoter
     primer_bind     4123..4139
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     mobile_element  4842..5888
                     /mobile_element_type="transposon:Piggybac"
                     /label=bac
     enhancer        6005..6487
                     /label=hr5 enhancer
                     /note="baculovirus early transcription enhancer"
     CDS             7334..8008
                     /label=DsRed2
                     /note="improved tetrameric variant of DsRed fluorescent
                     protein"
     CDS             8018..8038
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     polyA_signal    8179..8300
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(8345..8615)
                     /label=tetracycline response element
                     /note="contains seven copies of the tetracycline operator
                     tetO"
ORIGIN
        1 ggtaccctgg cgtctaattg gggtaagggt accctggcgt ctaattgggg taagttttcc
       61 cgttcttttc tgggttcttc ccttttgctc atccttgctg cactaccttc aggtgcaagt
      121 aaagccatac agtgaaccgc gggatcgccc accatgctgc tggtcaacca gtcgcaccag
      181 ggattcaaca aagaacacac ctcgaagatg gtgtcggcca tcgtgctgta cgtgctgctg
      241 gccgctgccg cccactcggc cttcgccaag aagaacggat acgccgtcga ttcgtcggga
      301 aaggccccgg aatgcctgct gtcgaactac tgcaacaacc aatgtaccaa ggtgcactac
      361 gccgataagg gatactgctg cctgctgagc tgctattgct tcggactgaa cgatgacaag
      421 aaggtgctgg aaatctcgga tacccgcaag tcgtactgcg ataccaccat catcaactga
      481 gcatgcgggc ccggccaaag agtctaattt ttgttcatca atgggttata acatatgggt
      541 tatattataa gtttgtttta agtttttgag actgataaga atgtttcgat cgaatattcc
      601 atagaacaac aatagtatta cctaattacc aagtcttaat ttagcaaaaa tgttattgct
      661 tatagaaaaa ataaattatt tattcccggg ccgatctgac aatgttcagt gcagagactc
      721 ggctacgcct cgtggacttt gaagttgacc aacaatgttt attcttacct ctaatagtcc
      781 tctgtggcaa ggtcaagatt ctgttagaag ccaatgaaga acctggttgt tcaataacat
      841 tttgttcgtc taatatttca ctaccgcttg acgttggctg cacttcatgt acctcatcta
      901 taaacgcttc ttctgtatcg ctctggacgt catcttcact tacgtgatct gatatttcac
      961 tgtcagaatc ctcaccaaca agctcgtcat cgctttgcag aagagcagag aggatatgct
     1021 catcgtctaa agaactaccc attttattat atattagtca cgatatctat aacaagaaaa
     1081 tatatatata ataagttatc acgtaagtag aacatgaaat aacaatataa ttatcgtatg
     1141 agttaaatct taaaagtcac gtaaaagata atcatgcgtc attttgactc acgcggtcgt
     1201 tatagttcaa aatcagtgac acttaccgca ttgacaagca cgcctcacgg gagctccaag
     1261 cggcgactga gatgtcctaa atgcacagcg acggattcgc gctatttaga aagagagagc
     1321 aatatttcaa gaatgcatgc gtcaatttta cgcagactat ctttctaggg ttaaaaaaga
     1381 tttgcgcttt actcgaccta aactttaaac acgtcataga atcttcgttt gacaaaaacc
     1441 acattgtggc caagctgtgt gacgcgacgc gcgctaaaga atggcaaacc aagtcgcgcg
     1501 agcgtcgaaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     1561 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     1621 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     1681 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     1741 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     1801 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     1861 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     1921 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     1981 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2041 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2101 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     2161 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2221 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2281 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2341 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     2401 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2461 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     2521 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     2581 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     2641 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     2701 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     2761 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     2821 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     2881 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     2941 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     3001 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     3061 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     3121 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     3181 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     3241 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     3301 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     3361 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     3421 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     3481 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     3541 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     3601 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     3661 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     3721 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     3781 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     3841 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     3901 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     3961 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
     4021 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     4081 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     4141 ccaagctttg tttaaaatat aacaaaattg tgatcccaca aaatgaagtg gggcaaaatc
     4201 aaataattaa tagtgtccgt aaacttgttg gtcttcaact ttttgaggaa cacgttggac
     4261 ggcaaatccg tgactataac acaagttgat ttaataattt tagccaacac gtcgggctgc
     4321 gtgttttttg ccgacgcgtc tgtgtacacg ttgattaact ggtcgattaa actgttgaaa
     4381 taatttaatt tttggttctt ctttaaatct gtgatgaaat tttttaaaat aactttaaat
     4441 tcttcattgg taaaaaatgc cacgttttgc aacttgtgag ggtctaatat gaggtcaaac
     4501 tcagtaggag ttttatccaa aaaagaaaac atgattacgt ctgtacacga acgcgtatta
     4561 acgcagagtg caaagtataa gagggttaaa aaatatattt tacgcaccat atacgcatcg
     4621 ggttgatatc gttaatatgg atcaatttga acagttgatt aacgtgtctc tgctcaagtc
     4681 tttgatcaaa acgcaaatcg acgaaaatgt gtcggacaat atcaagtcga tgagcgaaaa
     4741 actaaaaagg ctagaatacg acaatctcac agacagcgtt gagatatacg gtattcacga
     4801 cagcaggctg aataataaaa aaattagaaa ctattattta accctagaaa gataatcata
     4861 ttgtgacgta cgttaaagat aatcatgcgt aaaattgacg catgtgtttt atcggtctgt
     4921 atatcgaggt ttatttatta atttgaatag atattaagtt ttattatatt tacacttaca
     4981 tactaataat aaattcaaca aacaatttat ttatgtttat ttatttatta aaaaaaaaca
     5041 aaaactcaaa atttcttcta taaagtaaca aaacttttaa acattctctc ttttacaaaa
     5101 ataaacttat tttgtacttt aaaaacagtc atgttgtatt ataaaataag taattagctt
     5161 aacttataca taatagaaac aaattatact tattagtcag tcagaaacaa ctttggcaca
     5221 tatcaatatt atgctctcga caaataactt ttttgcattt tttgcacgat gcatttgcct
     5281 ttcgccttat tttagagggg cagtaagtac agtaagtacg ttttttcatt actggctctt
     5341 cagtactgtc atctgatgta ccaggcactt catttggcaa aatattagag atattatcgc
     5401 gcaaatatct cttcaaagta ggagcttcta aacgcttacg cataaacgat gacgtcaggc
     5461 tcatgtaaag gtttctcata aattttttgc gactttgaac cttttctccc ttgctactga
     5521 cattatggct gtatataata aaagaattta tgcaggcaat gtttatcatt ccgtacaata
     5581 atgccatagg ccacctattc gtcttcctac tgcaggtcat cacagaacac atttggtcta
     5641 gcgtgtccac tccgccttta gtttgattat aatacataac catttgcggt ttaccggtac
     5701 tttcgttgat agaagcatcc tcatcacaag atgataataa gtataccatc ttagctggct
     5761 tcggtttata tgagacgaga gtaaggggtc cgtcaaaaca aaacatcgat gttcccactg
     5821 gcctggagcg actgtttttc agtacttccg gtatctcgcg tttgtttgat cgcacggttc
     5881 ccacaatggt tgcggccggc cagatttaaa tgagcggccg catggtaccc attgcttgtc
     5941 atttattaat ttggatgatg tcatttgttt ttaaaattga actggcttta cgagtagaat
     6001 tctacgcgta aaacacaatc aagtatgagt cataagctga tgtcatgttt tgcacacggc
     6061 tcataaccga actggcttta cgagtagaat tctacttgta acgcacgatc agtggatgat
     6121 gtcatttgtt tttcaaatcg agatgatgtc atgttttgca cacggctcat aaactcgctt
     6181 tacgagtaga attctacgtg taacgcacga tcgattgatg agtcatttgt tttgcaatat
     6241 gatatcatac aatatgactc atttgttttt caaaaccgaa cttgatttac gggtagaatt
     6301 ctacttgtaa agcacaatca aaaagatgat gtcatttgtt tttcaaaact gaactcgctt
     6361 tacgagtaga attctacgtg taaaacacaa tcaagaaatg atgtcatttg ttataaaaat
     6421 aaaagctgat gtcatgtttt gcacatggct cataactaaa ctcgctttac gggtagaatt
     6481 ctacgcgtaa aacatgattg ataattaaat aattcatttg caagctatac gttaaatcaa
     6541 acggacgctc gaggttgcac aacactatta tcgatttgca gttcgggaca taaatgttta
     6601 aatatatcga tgtctttgtg atgcgcgcga catttttgta ggttattgat aaaatgaacg
     6661 gatacgttgc ccgacattat cattaaatcc ttggcgtaga atttgtcggg tccattgtcc
     6721 gtgtgcgcta gcatgcccgt aacggacctc gtacttttgg cttcaaaggt tttgcgcaca
     6781 gacaaaatgt gccacacttg cagctctgca tgtgtgcgcg ttaccacaaa tcccaacggc
     6841 gcagtgtact tgttgtatgc aaataaatct cgataaaggc gcggcgcgcg aatgcagctg
     6901 atcacgtacg ctcctcgtgt tccgttcaag gacggtgtta ccgacctcag attaatgttt
     6961 atcggccgac tgttttcgta tccgctcacc aaacgcgttt ttgcattaac attgtatgtc
     7021 ggcggatgtt ctatatctaa tttgaataaa taaacgataa ccgcgttggt tttagagggc
     7081 ataataaaag aaatattgtt atcgtgttcg ccattagggc agtataaatt gacgttcatg
     7141 ttggatattg tttcagttgc aagttgacac tggcggcgac aagacaattc taattggggt
     7201 aagttttccc gttcttttct gggttcttcc cttttgctca tccttgctgc actaccttca
     7261 ggtgcaagtt gagattcagg ccaccatggg agatcccacc ccacccaaga agaagcgcaa
     7321 accggtcgcc accatggcct cctccgagaa cgtcatcacc gagttcatgc gcttcaaggt
     7381 gcgcatggag ggcaccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
     7441 cccctacgag ggccacaaca ccgtgaagct gaaggtgacc aagggcggcc ccctgccctt
     7501 cgcctgggac atcctgtccc cccagttcca gtacggctcc aaggtgtacg tgaagcaccc
     7561 cgccgacatc cccgactaca agaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     7621 gatgaacttc gaggacggcg gcgtggcgac cgtgacccag gactcctccc tgcaggacgg
     7681 ctgcttcatc tacaaggtga agttcatcgg cgtgaacttc ccctccgacg gccccgtgat
     7741 gcagaagaag accatgggct gggaggcctc caccgagcgc ctgtaccccc gcgacggcgt
     7801 gctgaagggc gagacccaca aggccctgaa gctgaaggac ggcggccact acctggtgga
     7861 gttcaagtcc atctacatgg ccaagaagcc cgtgcagctg cccggctact actacgtgga
     7921 cgccaagctg gacatcacct cccacaacga ggactacacc atcgtggagc agtacgagcg
     7981 caccgagggc cgccaccacc tgttcctgag atctcgaccc aagaaaaagc ggaaggtgga
     8041 ggacccgtaa gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta
     8101 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     8161 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     8221 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgttc
     8281 aaactcatca atgtatctta acgcgagtta attaagaggc gcgccaggtt tcgactttca
     8341 cttttctcta tcactgatag ggagtggtaa actcgacttt cacttttctc tatcactgat
     8401 agggagtggt aaactcgact ttcacttttc tctatcactg atagggagtg gtaaactcga
     8461 ctttcacttt tctctatcac tgatagggag tggtaaactc gactttcact tttctctatc
     8521 actgataggg agtggtaaac tcgactttca cttttctcta tcactgatag ggagtggtaa
     8581 actcgacttt cacttttctc tatcactgat agggagtggt aaactcgaaa acgagcgccg
     8641 gagtataaat agaggcgctt cgtctacgga gcgacaattc aattcaaaca agcaaagtga
     8701 acacgtcgct aagcgaaagc taagcaaata aacaagcgca gctgaacaag ctaaacaatc
     8761 tgc
//
