LOCUS       62056_301        6610 bp DNA     circular SYN 12-AUG-2020
DEFINITION  Cloning vector AGG1221:pCHIKVLuc, complete sequence.
ACCESSION   MT119958
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6610)
  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 6610)
  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 6610)
  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..6610
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        47..529
                     /label=hr5 enhancer
                     /note="baculovirus early transcription enhancer"
     intron          1273..1342
                     /note="adh"
     misc_feature    1345..1353
                     /label=Kozak sequence
                     /note="Kozak sequence"
     misc_feature    1354..1626
                     /gene="CHIKV-Ubi-luciferase polyprotein"
                     /label=CHIKV nsP2
                     /note="CHIKV nsP2"
     CDS             1627..1851
                     /label=ubiquitin-K48R
                     /note="human ubiquitin variant in which Lys-48 has been
                     mutated to arginine"
     CDS             1852..3501
                     /label=luciferase
                     /note="firefly luciferase"
     polyA_signal    complement(3545..3666)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4085..4673)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4847..5704)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5705..5809)
                     /label=AmpR promoter
     rep_origin      5836..6291
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     polyA_signal    6422..6470
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     misc_feature    6484..6575
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from
                     the human alpha-2 globin gene"
ORIGIN
        1 gggctcgaga tctgcgatct aagtaagctt tacgagtaga attctacgcg taaaacacaa
       61 tcaagtatga gtcataatct gatgtcatgt tttgtacacg gctcataacc gaactggctt
      121 tacgagtaga attctacttg taatgcacga tcagtggatg atgtcatttg tttttcaaat
      181 cgagatgatg tcatgttttg cacacggctc ataaactcgc tttacgagta gaattctacg
      241 tgtaacgcac gatcgattga tgagtcattt gttttgcaat atgatatcat acaatatgac
      301 tcatttgttt ttcaaaaccg aacttgattt acgggtagaa ttctacttgt aaagcacaat
      361 caaaaagatg atgtcatttg tttttcaaaa ctgaactcgc tttacgagta gaattctacg
      421 tgtaaaacac aatcaagaaa tgatgtcatt tgttataaaa ataaaagctg atgtcatgtt
      481 ttgcacatgg ctcataacta aactcgcttt acgggtagaa ttctacgcgt aaaacatgat
      541 tgataattaa ataattcatt tgcaagctat acgttaaatc aaacggacgc tcgaggttgc
      601 acaacactat tatcgatttg cagttcggga cataaatgtt taaatatatc gatgtctttg
      661 tgatgcgcgc gacatttttg taggttattg ataaaatgaa cggatacgtt gcccgacatt
      721 atcattaaat ccttggcgta gaatttgtcg ggtccattgt ccgtgtgcgc tagtagcatg
      781 cccgtaacgg acctcgtact tttggcttca aaggttttgc gcacagacaa aatgtgccac
      841 acttgcagct ctgcatgtgt gcgcgttacc acaaatccca acggcgcagt gtacttgttg
      901 tatgcaaata aatctcgata aaggcgcggc gcgcgaatgc agctgatcac gtacgctcct
      961 cgtgttccgt tcaaggacgg tgttatcgac ctcagattaa tgtttatcgg ccgactgttt
     1021 tcgtatccgc tcaccaaacg cgtttttgca ttaacattgt atgtcggcgg atgttctata
     1081 tctaatttga ataaataaac gataaccgcg ttggttttag agggcataat aaaagaaata
     1141 ttgttatcgt gttcgccatt agggcagtat aaattgacgt tcatgttgga tattgtttca
     1201 gttgcaagtt gacactggcg gcgacaagca attggtaccc gggtaggatc ctagtgaatt
     1261 cctaatctgg cggtaagttg atcaaaggaa acgcaaagtt ttcaagaaaa aacaaaacta
     1321 atttgattta taacaccttt agaagccgcc accatgaaag ttgtactttg tggtgacccg
     1381 aagcagtgcg gcttcttcaa tatgatgcag atgaaagtca actataatca caacatctgc
     1441 acccaagtgt accacaaaag tatctccagg cggtgtacac tgcctgtgac cgccattgtg
     1501 tcatcgttgc attacgaagg caaaatgcgc actacgaatg agtacaacaa gccgattgta
     1561 gtggacacta caggctcaac aaaacctgat cctggagacc tcgtgttaac gtgcttcaga
     1621 gggtggcaga ttttcgtcaa gaccctgacc ggcaagacca tcaccctgga ggtggagccg
     1681 agcgatacca tcgagaacgt gaaggccaag atccaggaca aggagggcat cccgccggat
     1741 cagcagcgct tgatcttcgc cggacgccag ctggaggatg gccgcaccct gagcgactac
     1801 aacatccaga aggagagcac cctgcatctg gtgctgcgcc tgcgcggtgg tatggaagac
     1861 gccaaaaaca taaagaaagg cccggcgcca ttctatccgc tggaagatgg aaccgctgga
     1921 gagcaactgc ataaggctat gaagagatac gccctggttc ctggaacaat tgcttttaca
     1981 gatgcacata tcgaggtgga catcacttac gctgagtact tcgaaatgtc cgttcggttg
     2041 gcagaagcta tgaaacgata tgggctgaat acaaatcaca gaatcgtcgt atgcagtgaa
     2101 aactctcttc aattctttat gccggtgttg ggcgcgttat ttatcggagt tgcagttgcg
     2161 cccgcgaacg acatttataa tgaacgtgaa ttgctcaaca gtatgggcat ttcgcagcct
     2221 accgtggtgt tcgtttccaa aaaggggttg caaaaaattt tgaacgtgca aaaaaagctc
     2281 ccaatcatcc aaaaaattat tatcatggat tctaaaacgg attaccaggg atttcagtcg
     2341 atgtacacgt tcgtcacatc tcatctacct cccggtttta atgaatacga ttttgtgcca
     2401 gagtccttcg atagggacaa gacaattgca ctgatcatga actcctctgg atctactggt
     2461 ctgcctaaag gtgtcgctct gcctcataga actgcctgcg tgagattctc gcatgccaga
     2521 gatcctattt ttggcaatca aatcattccg gatactgcga ttttaagtgt tgttccattc
     2581 catcacggtt ttggaatgtt tactacactc ggatatttga tatgtggatt tcgagtcgtc
     2641 ttaatgtata gatttgaaga agagctgttt ctgaggagcc ttcaggatta caagattcaa
     2701 agtgcgctgc tggtgccaac cctattctcc ttcttcgcca aaagcactct gattgacaaa
     2761 tacgatttat ctaatttaca cgaaattgct tctggtggcg ctcccctctc taaggaagtc
     2821 ggggaagcgg ttgccaagag gttccatctg ccaggtatca ggcaaggata tgggctcact
     2881 gagactacat cagctattct gattacaccc gagggggatg ataaaccggg cgcggtcggt
     2941 aaagttgttc cattttttga agcgaaggtt gtggatctgg ataccgggaa aacgctgggc
     3001 gttaatcaaa gaggcgaact gtgtgtgaga ggtcctatga ttatgtccgg ttatgtaaac
     3061 aatccggaag cgaccaacgc cttgattgac aaggatggat ggctacattc tggagacata
     3121 gcttactggg acgaagacga acacttcttc atcgttgacc gcctgaagtc tctgattaag
     3181 tacaaaggct atcaggtggc tcccgctgaa ttggaatcca tcttgctcca acaccccaac
     3241 atcttcgacg caggtgtcgc aggtcttccc gacgatgacg ccggtgaact tcccgccgcc
     3301 gttgttgttt tggagcacgg aaagacgatg acggaaaaag agatcgtgga ttacgtcgcc
     3361 agtcaagtaa caaccgcgaa aaagttgcgc ggaggagttg tgtttgtgga cgaagtaccg
     3421 aaaggtctta ccggaaaact cgacgcaaga aaaatcagag agatcctcat aaaggccaag
     3481 aagggcggaa agatcgccgt gtaattctag agtcggggcg gccggccgct tcgagcagac
     3541 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     3601 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     3661 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     3721 gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tccgtcgacc
     3781 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat
     3841 cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc
     3901 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3961 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4021 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4081 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4141 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4201 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4261 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4321 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4381 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4441 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4501 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4561 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4621 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4681 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4741 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4801 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4861 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4921 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4981 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5041 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5101 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5161 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5221 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5281 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5341 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5401 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5461 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5521 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5581 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5641 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5701 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5761 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5821 gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     5881 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     5941 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     6001 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     6061 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     6121 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     6181 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     6241 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt tgccattcgc
     6301 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     6361 agcccaagct accatgataa gtaagtaata ttaaggtacg ggaggtactt ggagcggccg
     6421 caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagt
     6481 actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa aataggctgt
     6541 ccccagtgca agtgcaggtg ccagaacatt tctctatcga taggtaccga gctcttacgc
     6601 gtgctagccc
//
