LOCUS       62056_296        6075 bp DNA     circular SYN 12-AUG-2020
DEFINITION  Cloning vector AGG1201:pZsG, complete sequence.
ACCESSION   MT119955
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6075)
  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 6075)
  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 6075)
  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..6075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    41..74
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(414..430)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(438..454)
                     /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(462..492)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(507..528)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(816..1404)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1578..2435)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(2436..2540)
                     /label=AmpR promoter
     enhancer        2912..3394
                     /label=hr5 enhancer
                     /note="baculovirus early transcription enhancer"
     intron          4138..4207
                     /note="adh"
     misc_feature    4228..4237
                     /label=Kozak sequence
                     /note="Kozak sequence"
     regulatory      4246..4269
                     /gene="ZsGreen"
                     /label=nuclear localization signal
                     /note="nuclear localization signal"
                     /regulatory_class="other"
     CDS             4285..4977
                     /label=ZsGreen1
                     /note="Zoanthus green fluorescent protein"
     CDS             4996..5016
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     3'UTR           5051..5832
                     /label=K10
                     /note="K10"
     polyA_signal    complement(5841..5962)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 ggcatatgtg tttaaaccga ggtacctgat ttggcgcgcc ataacttcgt ataatgtatg
       61 ctatacgaag ttatttgata tctataacaa gaaaatatat atataataag ttatcacgta
      121 agtagaacat gaaataacaa tataattatc gtatgagtta aatcttaaaa gtcacgtaaa
      181 agataatcat gcgtcatttt gactcacgcg gtcgttatag ttcaaaatca gtgacactta
      241 ccgcattgac aagcacgcct cacgggagct ccaagcggcg actgagatgt cctaaatgca
      301 cagcgacgga ttcgcgctat ttagaaagag agagcaatat ttcaagaatg catgcgtcaa
      361 ttttacgcag actatctttc tagggttaat ctagcgagct tggcgtaatc atggtcatag
      421 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
      481 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
      541 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
      601 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
      661 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
      721 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
      781 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
      841 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
      901 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
      961 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     1021 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     1081 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     1141 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     1201 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     1261 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     1321 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     1381 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     1441 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     1501 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     1561 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     1621 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     1681 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     1741 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     1801 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     1861 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     1921 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     1981 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     2041 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     2101 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     2161 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     2221 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     2281 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     2341 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     2401 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     2461 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     2521 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc caccagctag attaacccta
     2581 gaaagataat catattgtga cgtacgttaa agataatcat gcgtaaaatt gacgcatgtg
     2641 ttttatcggt ctgtatatcg aggtttattt attaatttga atagatatta agttttatta
     2701 tatttacact tacatactaa taataaattc aacaaacaat ttatttatgt ttatttattt
     2761 attaaaaaaa aacaaaaact caaaatttct tctataaagt aacaaaactt ttatcgaatt
     2821 gcttcggcgc caagtagtgc cccaactggg gtaacctttg agttctctca gttgggggcg
     2881 taggtcgaca agctttacga gtagaattct acgcgtaaaa cacaatcaag tatgagtcat
     2941 aatctgatgt catgttttgt acacggctca taaccgaact ggctttacga gtagaattct
     3001 acttgtaatg cacgatcagt ggatgatgtc atttgttttt caaatcgaga tgatgtcatg
     3061 ttttgcacac ggctcataaa ctcgctttac gagtagaatt ctacgtgtaa cgcacgatcg
     3121 attgatgagt catttgtttt gcaatatgat atcatacaat atgactcatt tgtttttcaa
     3181 aaccgaactt gatttacggg tagaattcta cttgtaaagc acaatcaaaa agatgatgtc
     3241 atttgttttt caaaactgaa ctcgctttac gagtagaatt ctacgtgtaa aacacaatca
     3301 agaaatgatg tcatttgtta taaaaataaa agctgatgtc atgttttgca catggctcat
     3361 aactaaactc gctttacggg tagaattcta cgcgtaaaac atgattgata attaaataat
     3421 tcatttgcaa gctatacgtt aaatcaaacg gacgctcgag gttgcacaac actattatcg
     3481 atttgcagtt cgggacataa atgtttaaat atatcgatgt ctttgtgatg cgcgcgacat
     3541 ttttgtaggt tattgataaa atgaacggat acgttgcccg acattatcat taaatccttg
     3601 gcgtagaatt tgtcgggtcc attgtccgtg tgcgctagta gcatgcccgt aacggacctc
     3661 gtacttttgg cttcaaaggt tttgcgcaca gacaaaatgt gccacacttg cagctctgca
     3721 tgtgtgcgcg ttaccacaaa tcccaacggc gcagtgtact tgttgtatgc aaataaatct
     3781 cgataaaggc gcggcgcgcg aatgcagctg atcacgtacg ctcctcgtgt tccgttcaag
     3841 gacggtgtta tcgacctcag attaatgttt atcggccgac tgttttcgta tccgctcacc
     3901 aaacgcgttt ttgcattaac attgtatgtc ggcggatgtt ctatatctaa tttgaataaa
     3961 taaacgataa ccgcgttggt tttagagggc ataataaaag aaatattgtt atcgtgttcg
     4021 ccattagggc agtataaatt gacgttcatg ttggatattg tttcagttgc aagttgacac
     4081 tggcggcgac aagcaattgg tacccgggta ggatcctagt gaattcctaa tctggcggta
     4141 agttgatcaa aggaaacgca aagttttcaa gaaaaaacaa aactaatttg atttataaca
     4201 cctttagaaa gcgaagttga gattcaggcc accatgggag atcccacccc acccaagaag
     4261 aagcgcaaag ctagctcggc caccatggcc cagtccaagc acggcctgac caaggagatg
     4321 accatgaagt accgcatgga gggctgcgtg gacggccaca agttcgtgat cacaggcgag
     4381 ggcatcggct accccttcaa gggcaagcag gccatcaacc tgtgcgtggt ggagggcggc
     4441 cccttgccct tcgccgagga catcttgtcc gccgccttca tgtacggcaa ccgcgtgttc
     4501 accgagtacc cccaggacat cgtggactac ttcaagaact cctgccccgc cggctacacc
     4561 tgggaccgct ccttcctgtt cgaggacggc gctgtgtgca tctgcaacgc cgacatcacc
     4621 gtgagcgtgg aggagaactg catgtaccac gagtccaagt tctacggcgt gaacttcccc
     4681 gccgacggcc ccgtgatgaa gaagatgacc gacaactggg agccctcctg cgagaagatc
     4741 atccccgtgc ccaagcaagg catcttgaag ggcgacgtta gcatgtacct gctgctgaag
     4801 gacggtggcc gcttgcgctg ccagttcgac accgtctaca aggccaagtc cgtgccccgc
     4861 aagatgcccg actggcactt catccagcac aagctgaccc gcgaggaccg cagcgacgcc
     4921 aagaaccaga agtggcacct gaccgagcac gccatcgcct ccggctccgc cttgccctcc
     4981 ggactcagat ctcgacccaa gaaaaagcgg aaggtggagg acccgtaaga tccaccggat
     5041 ctagataact ggagcttgat aacattatac ctaaacccat ggtcaagagt aaacatttct
     5101 gcctttgaag ttgagaacac aattaagcat cccctggtta aacctgacat tcatacttgt
     5161 taatagcgcc ataaacatag caccaatttc gaagaaatca gttaaaagca attagcaatt
     5221 agcaattagc aataactctg ctgacttcaa aacgagaaga gttgcaagta tttgtaaggc
     5281 acagtttata gaccaccgac ggctcattag ggctcgtcat gtaactaagc gcggtgaaac
     5341 ccaattgaac atatagtgga attattatta tcaatgggga agatttaacc ctcaggtagc
     5401 aaagtaattt aattgcaaat agagagtcct aagactaaat aatatattta aaaatctggc
     5461 cctttgacct tgcttgtcag gtgcatttgg gttcaatcgt aagttgcttc tatataaaca
     5521 ctttccccat ccccgcaata atgaagaata ccgcagaata aagagagatt tgcaacaaaa
     5581 aataaaggca ttgcgaaaac tttttatggg ggatcattac actcgggcct acggttacaa
     5641 ttcccagcca cttaagcgac aagtttggcc aacaatccat ctaatagcta atagcgcaat
     5701 cactggtaat cgcaagagta tataggcaat agaacccatg gatttgacca aaggtaaccg
     5761 agacaatgga gaagcaagag gatttcaaac tgaacaccca cagtactgtg tactaccact
     5821 ggcgcgtttg ggggccggcc taagatacat tgatgagttt ggacaaacca caactagaat
     5881 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     5941 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     6001 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga
     6061 ttatgatcaa ttaat
//
