LOCUS       62056_291        6112 bp DNA     circular SYN 12-AUG-2020
DEFINITION  Cloning vector AGG1186:pHr5Fluc, complete sequence.
ACCESSION   MT119956
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6112)
  AUTHORS   Tng PYL., Carabajal Paladino L, Verkuijl SAN., Purcell J, Merits A, 
            Leftwich PT, Fragkoudis R, Noad R, Alphey L.
  TITLE     Cas13b-dependent and Cas13b-independent RNA knockdown of viral 
            sequences in mosquito cells following guide RNA expression
  JOURNAL   Commun Biol 3 (1), 413 (2020)
  PUBMED    32737398
REFERENCE   2  (bases 1 to 6112)
  AUTHORS   Tng PYL., Carabajal Paladino L, Verkuijl S, Purcell J, Merits A, 
            Leftwich PT, Fragkoudis R, Noad R, Alphey L.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-FEB-2020) Arthropod Genetics Group, The Pirbright 
            Institute, Ash Road, Pirbright, Woking, Surrey GU24 0NF, UK
REFERENCE   3  (bases 1 to 6112)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Commun 
            Biol"; date: "2020"; volume: "3"; issue: "1"; pages: "413"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (26-FEB-2020) Arthropod Genetics Group, The Pirbright Institute, Ash
            Road, Pirbright, Woking, Surrey GU24 0NF, UK"
FEATURES             Location/Qualifiers
     source          1..6112
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        134..616
                     /label=hr5 enhancer
                     /note="baculovirus early transcription enhancer"
     intron          1360..1429
                     /note="adh"
     misc_feature    1432..1440
                     /label=Kozak sequence
                     /note="Kozak sequence"
     CDS             1441..3090
                     /label=luciferase
                     /note="firefly luciferase"
     polyA_signal    complement(3134..3255)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3674..4262)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4436..5293)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5294..5398)
                     /label=AmpR promoter
     rep_origin      5425..5880
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     polyA_signal    6011..6059
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     misc_feature    6073..6112
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from
                     the human alpha-2 globin gene"
ORIGIN
        1 ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct ctatcgatag
       61 gtaccgagct cttacgcgtg ctagcccggg ctcgagatct gcgatctaag taagctttac
      121 gagtagaatt ctacgcgtaa aacacaatca agtatgagtc ataatctgat gtcatgtttt
      181 gtacacggct cataaccgaa ctggctttac gagtagaatt ctacttgtaa tgcacgatca
      241 gtggatgatg tcatttgttt ttcaaatcga gatgatgtca tgttttgcac acggctcata
      301 aactcgcttt acgagtagaa ttctacgtgt aacgcacgat cgattgatga gtcatttgtt
      361 ttgcaatatg atatcataca atatgactca tttgtttttc aaaaccgaac ttgatttacg
      421 ggtagaattc tacttgtaaa gcacaatcaa aaagatgatg tcatttgttt ttcaaaactg
      481 aactcgcttt acgagtagaa ttctacgtgt aaaacacaat caagaaatga tgtcatttgt
      541 tataaaaata aaagctgatg tcatgttttg cacatggctc ataactaaac tcgctttacg
      601 ggtagaattc tacgcgtaaa acatgattga taattaaata attcatttgc aagctatacg
      661 ttaaatcaaa cggacgctcg aggttgcaca acactattat cgatttgcag ttcgggacat
      721 aaatgtttaa atatatcgat gtctttgtga tgcgcgcgac atttttgtag gttattgata
      781 aaatgaacgg atacgttgcc cgacattatc attaaatcct tggcgtagaa tttgtcgggt
      841 ccattgtccg tgtgcgctag tagcatgccc gtaacggacc tcgtactttt ggcttcaaag
      901 gttttgcgca cagacaaaat gtgccacact tgcagctctg catgtgtgcg cgttaccaca
      961 aatcccaacg gcgcagtgta cttgttgtat gcaaataaat ctcgataaag gcgcggcgcg
     1021 cgaatgcagc tgatcacgta cgctcctcgt gttccgttca aggacggtgt tatcgacctc
     1081 agattaatgt ttatcggccg actgttttcg tatccgctca ccaaacgcgt ttttgcatta
     1141 acattgtatg tcggcggatg ttctatatct aatttgaata aataaacgat aaccgcgttg
     1201 gttttagagg gcataataaa agaaatattg ttatcgtgtt cgccattagg gcagtataaa
     1261 ttgacgttca tgttggatat tgtttcagtt gcaagttgac actggcggcg acaagcaatt
     1321 ggtacccggg taggatccta gtgaattcct aatctggcgg taagttgatc aaaggaaacg
     1381 caaagttttc aagaaaaaac aaaactaatt tgatttataa cacctttaga agccgccacc
     1441 atggaagacg ccaaaaacat aaagaaaggc ccggcgccat tctatccgct ggaagatgga
     1501 accgctggag agcaactgca taaggctatg aagagatacg ccctggttcc tggaacaatt
     1561 gcttttacag atgcacatat cgaggtggac atcacttacg ctgagtactt cgaaatgtcc
     1621 gttcggttgg cagaagctat gaaacgatat gggctgaata caaatcacag aatcgtcgta
     1681 tgcagtgaaa actctcttca attctttatg ccggtgttgg gcgcgttatt tatcggagtt
     1741 gcagttgcgc ccgcgaacga catttataat gaacgtgaat tgctcaacag tatgggcatt
     1801 tcgcagccta ccgtggtgtt cgtttccaaa aaggggttgc aaaaaatttt gaacgtgcaa
     1861 aaaaagctcc caatcatcca aaaaattatt atcatggatt ctaaaacgga ttaccaggga
     1921 tttcagtcga tgtacacgtt cgtcacatct catctacctc ccggttttaa tgaatacgat
     1981 tttgtgccag agtccttcga tagggacaag acaattgcac tgatcatgaa ctcctctgga
     2041 tctactggtc tgcctaaagg tgtcgctctg cctcatagaa ctgcctgcgt gagattctcg
     2101 catgccagag atcctatttt tggcaatcaa atcattccgg atactgcgat tttaagtgtt
     2161 gttccattcc atcacggttt tggaatgttt actacactcg gatatttgat atgtggattt
     2221 cgagtcgtct taatgtatag atttgaagaa gagctgtttc tgaggagcct tcaggattac
     2281 aagattcaaa gtgcgctgct ggtgccaacc ctattctcct tcttcgccaa aagcactctg
     2341 attgacaaat acgatttatc taatttacac gaaattgctt ctggtggcgc tcccctctct
     2401 aaggaagtcg gggaagcggt tgccaagagg ttccatctgc caggtatcag gcaaggatat
     2461 gggctcactg agactacatc agctattctg attacacccg agggggatga taaaccgggc
     2521 gcggtcggta aagttgttcc attttttgaa gcgaaggttg tggatctgga taccgggaaa
     2581 acgctgggcg ttaatcaaag aggcgaactg tgtgtgagag gtcctatgat tatgtccggt
     2641 tatgtaaaca atccggaagc gaccaacgcc ttgattgaca aggatggatg gctacattct
     2701 ggagacatag cttactggga cgaagacgaa cacttcttca tcgttgaccg cctgaagtct
     2761 ctgattaagt acaaaggcta tcaggtggct cccgctgaat tggaatccat cttgctccaa
     2821 caccccaaca tcttcgacgc aggtgtcgca ggtcttcccg acgatgacgc cggtgaactt
     2881 cccgccgccg ttgttgtttt ggagcacgga aagacgatga cggaaaaaga gatcgtggat
     2941 tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt gtttgtggac
     3001 gaagtaccga aaggtcttac cggaaaactc gacgcaagaa aaatcagaga gatcctcata
     3061 aaggccaaga agggcggaaa gatcgccgtg taattctaga gtcggggcgg ccggccgctt
     3121 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     3181 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     3241 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     3301 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat
     3361 ccgtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
     3421 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
     3481 gccggcagcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3541 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3601 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3661 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3721 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3781 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3841 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3901 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3961 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4021 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4081 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4141 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4201 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4261 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4321 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4381 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4441 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4501 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     4561 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4621 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4681 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4741 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4801 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4861 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4921 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4981 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5041 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5101 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5161 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5221 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5281 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5341 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5401 catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     5461 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     5521 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     5581 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     5641 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     5701 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     5761 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     5821 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     5881 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     5941 tattacgcca gcccaagcta ccatgataag taagtaatat taaggtacgg gaggtacttg
     6001 gagcggccgc aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga
     6061 atcgatagta ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aa
//
