LOCUS       sequence-361375-        3477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gRNA-expressing plasmid under Aedres aegypti AAEL017774 U6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3477)
  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 3477)
  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-AeU6-2gRNA
FEATURES             Location/Qualifiers
     source          1..3477
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     623..642
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     674..690
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     674..690
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1244..1260)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1245..1261)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(1289..1307)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1328..1344)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1328..1344)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1341..1363)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1352..1368
                     /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)."
     promoter        complement(join(1376..1382,1383..1400,1401..1406))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1421..1442
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1559..1576)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1730..2318)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1810..1829)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2489..3280,3281..3349))
                     /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     3112..3131
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3350..3454)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg agctccaccg
      661 cggtggcggc cgctctagaa ctagtggatc ccccggtctc actctaattg gagctgccag
      721 acaaatttga tttgtccgtg cggtatgcat atgtacctac ttacgtctgt cgttttgtct
      781 ccgtttaccg ggagggaaag tctggaaaca tggaaactct atagttgcca ggtagaccat
      841 ctgcctccgt cggctggctg gattccaatt tgaatattgg ctaattggaa gagatggaag
      901 tttttgaatg gatgattgaa taattgaagc gactccgggt acctgtttgt aagctctgca
      961 acagtgccat agattcgtgt cagtccatca ctagaatcaa atcaacttgt acttgatata
     1021 taagagcaga ggcaagagta gtgaaatgca gtcttcgaga agacctgttt cagagctatg
     1081 ctggaaacag catagcaagt tgaaataagg ctagtccgtt atcaacttga aaaagtggca
     1141 ccgagtcggt gcttttttta caaatctatt atgcatgagg tactatgttg gcagagtaat
     1201 gaattccaga gaccgggctg caggaattcg atatcaagct tatcgatacc gtcgacctcg
     1261 agggggggcc cggtaccagc ttttgttccc tttagtgagg gttaatttcg agcttggcgt
     1321 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     1381 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     1441 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     1501 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     1561 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     1621 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     1681 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1741 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1801 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1861 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1921 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1981 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2041 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2101 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2161 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2221 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2281 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2341 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     2401 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2461 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     2521 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     2581 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gatccacgct
     2641 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     2701 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     2761 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     2821 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     2881 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     2941 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3001 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3061 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     3121 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     3181 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     3241 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     3301 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     3361 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     3421 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccac
//
