LOCUS       sequence-361374-        3588 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gRNA-expressing plasmid under Aedes aegypti AAEL017702 U6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3588)
  AUTHORS   Lutrat C, Burckbuchler M, Olmo RP, Beugnon R, Fontaine A, Akbari OS,
            Argiles-Herrero R, Baldet T, Bouyer J, Marois E
  TITLE     Combining two genetic sexing strains allows sorting of 
            non-transgenic males for Aedes genetic control.
  JOURNAL   Commun Biol. 2023 Jun 16;6(1):646. doi: 10.1038/s42003-023-05030-7.
  PUBMED    37328568
REFERENCE   2  (bases 1 to 3588)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Commun 
            Biol."; date: "2023-06-16"; volume: "6(1):646. doi"; pages: " 
            10.1038/s42003-023-05030-7"
COMMENT     Sequence Label: pKSB-AeU6-1gRNA
FEATURES             Location/Qualifiers
     source          1..3588
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    39..60
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(75..80,81..98,99..105)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    113..129
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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     118..140
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     137..153
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     137..153
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        174..192
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     220..236
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     221..237
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(902..918)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(902..918)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(950..969)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(951..969)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(976..993)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(976..992)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(985..1007)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1134..1589
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1221..1240)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1431..1452
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1615..1719
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1720..1788,1789..2580)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1938..1957)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2751..3339
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3240..3259
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3493..3510
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
       61 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
      121 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct cgaaattaac
      181 cctcactaaa gggaacaaaa gctggtaccg ggccccccct cgaggtcgac ggtatcgata
      241 agcttgatat cgaattcctg cagcccggtc tcattccata gtaaatatac tacaaataat
      301 attaatgttc catgaaaaaa ggagtaagag tctggtaacc ctagtgcacg caaatatctc
      361 gcgggcatat ttggttgctg aggtatattt atatttgaac gccatgagaa aaagcggaag
      421 aaattggctc atggccgatt ttaaggatat ttaaaaattg tacaatgtac atataattaa
      481 acatccgttc cttcaatgtg ttcttttttt taagcgtgtg ttaaaagttt gctctggtgg
      541 tgaattcacg ctctacccgt tcaggcagca ttcatcgaaa agccctatct gctcgcacac
      601 atttacaaaa tgctgattgc gttgtgtgct gaatgggtca ctcgtccgtc actgcttgct
      661 gtgtacactg tacagttacg cagtctgtgc atcgctagaa tcatatttac ggaaaagtat
      721 tatatatacc caatgcgttg ctcatcggtt gcagtcttcg agaagacctg tttcagagct
      781 atgctggaaa cagcatagca agttgaaata aggctagtcc gttatcaact tgaaaaagtg
      841 gcaccgagtc ggtgcttttt ttgtggtttt attattcgat aattgtggat agagaccggg
      901 ggatccacta gttctagagc ggccgccacc gcggtggagc tccaattcgc cctatagtga
      961 gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     1021 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     1081 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
     1141 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     1201 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     1261 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     1321 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     1381 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     1441 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     1501 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     1561 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     1621 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     1681 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     1741 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     1801 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     1861 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     1921 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     1981 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2041 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2101 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2161 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2221 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     2281 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2341 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     2401 tttattgctg ataaatctgg agccggtgag cgtggatctc gcggtatcat tgcagcactg
     2461 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     2521 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     2581 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     2641 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     2701 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2761 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2821 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2881 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     2941 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3001 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3061 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3121 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3181 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3241 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3301 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3361 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3421 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3481 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     3541 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacagg
//
