LOCUS       sequence-221209-        7810 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses gRNA in Aedes aegypti.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7810)
  AUTHORS   Li M, Yang T, Kandul NP, Bui M, Gamez S, Raban R, Bennett J, Sanchez
            C HM, Lanzaro GC, Schmidt H, Lee Y, Marshall JM, Akbari OS
  TITLE     Development of a confinable gene drive system in the human disease 
            vector Aedes aegypti.
  JOURNAL   Elife. 2020 Jan 21;9. pii: 51701. doi: 10.7554/eLife.51701.
  PUBMED    31960794
REFERENCE   2  (bases 1 to 7810)
  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: "Elife."; 
            date: "2020-01-21"; volume: "9. pii: 51701. doi"; pages: " 
            10.7554/eLife.51701"
COMMENT     Sequence Label: U6d-T
FEATURES             Location/Qualifiers
     source          1..7810
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(95..683)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(175..194)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(854..1645,1646..1714))
                     /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     1477..1496
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1715..1819)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1887..1905
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1943..1965)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2065..2084
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2278..2300
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2292..2309
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2293..2309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   2998..3060
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     regulatory      4433..4442
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4439..4441,4442..4444,4445..5158)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4484..4505)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4745..4764)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5092..5113
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5281..5402
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5318..5337)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5372..5391
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    6365..6434
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     repeat_region   7287..7321
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7503..7519)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7503..7519)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7516..7538)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7527..7543
                     /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(7551..7557,7558..7575,7576..7581))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7596..7617
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7734..7751)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
       61 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
      121 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
      181 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
      241 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct
      301 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
      361 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
      421 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
      481 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
      541 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
      601 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
      661 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
      721 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
      781 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
      841 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
      901 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
      961 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     1021 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     1081 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     1141 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     1201 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     1261 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     1321 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     1381 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     1441 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     1501 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     1561 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     1621 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     1681 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     1741 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     1801 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     1861 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     1921 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     1981 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     2041 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     2101 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     2161 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     2221 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     2281 agtcacgacg ttgtaaaacg acggccagtg ccaagctttg tttaaaatat aacaaaattg
     2341 tgatcccaca aaatgaagtg gggcaaaatc aaataattaa ctagtgtccg taaacttgtt
     2401 ggtcttcaac tttttgagga acacgttgga cggcaaatcg tgactataac acaagttgat
     2461 ttaataattt tagccaacac gtcgggctgc gtgttttttg cgctctgtgt acacgttgat
     2521 taactggtcg attaaataat ttaatttttg gttcttcttt aaatctgtga tgaaattttt
     2581 taaaataact ttaaattctt cattggtaaa aaatgccacg ttttgcaact tgtgagggtc
     2641 taatatgagg tcaaactcag taggagtttt atccaaaaaa gaaaacatga ttacgtctgt
     2701 acacgaacgc gtattaacgc agagtgcaaa gtataagagg gttaaaaaat atattttacg
     2761 caccatatac gcatcgggtt gatatcgtta atatggatca atttgaacag ttgattaacg
     2821 tgtctctgct caagtctttg atcaaaacgc aaatcgacga aaatgtgtcg gacaatatca
     2881 agtcgatgag cgaaaaacta aaaaggctag aatacgacaa tctcacagac agcgttgaga
     2941 tatacggtat tcacgacagc aggctgaata ataaaaaaat tagaaactat tatttaaccc
     3001 tagaaagata atcatattgt gacgtacgtt aaagataatc atgcgtaaaa ttgacgcatg
     3061 tgttttatcg gtctgtatat cgaggtttat ttattaattt gaatagatat taagttttat
     3121 tatatttaca cttacatact aataataaat tcaacaaaca atttatttat gtttatttat
     3181 ttattaaaaa aaaacaaaaa ctcaaaattt cttctataaa gtaacaaaac ttttaaacat
     3241 tctctctttt acaaaaataa acttattttg tactttaaaa acagtcatgt tgtattataa
     3301 aataagtaat tagcttaact tatacataat agaaacaaat tatacttatt agtcagtcag
     3361 aaacaacttt ggcacatatc aatattatgc tctcgacaaa taactttttt gcattttttg
     3421 cacgatgcat ttgcctttcg ccttatttta gaggggcagt aagtacagta agtacgtttt
     3481 ttcattactg gctcttcagt actgtcatct gatgtaccag gcacttcatt tggcaaaata
     3541 ttagagatat tatcgcgcaa atatctcttc aaagtaggag cttctaaacg cttacgcata
     3601 aacgatgacg tcaggctcat gtaaaggttt ctcataaatt ttttgcgact ttgaaccttt
     3661 tctcccttgc tactgacatt atggctgtat ataataaaag aatttatgca ggcaatgttt
     3721 atcattccgt acaataatgc cataggccac ctattcgtct tcctactgca ggtcatcaca
     3781 gaacacattt ggtctagcgt gtccactccg cctttagttt gattataata cataaccatt
     3841 tgcggtttac cggtactttc gttgatagaa gcatcctcat cacaagatga taataagtat
     3901 accatcttag ctggcttcgg tttatatgag acgagagtaa ggggtccgtc aaaacaaaac
     3961 atcgatgttc ccactggcct ggagcgactg tttttcagta cttccggtat ctcgcgtttg
     4021 tttgatcgca cggttcccac aatggttaat tcgagctcgc ccgggcttaa gagttcccac
     4081 aatggttaat tcgagctcgc ccgggcttaa gagttcccac aatggttaat tcgagctcgc
     4141 ccgggcttaa gagcgcgccg ttcccacaat ggttaattcg agctcgcccg gggatctaat
     4201 tcaattagag actaattcaa ttagagctaa ttcaattagg atccaagctt atcgatttcg
     4261 aaccctcgac cgccggagta taaatagagg cgcttcgtct acggagcgac aattcaattc
     4321 aaacaagcaa agtgaacacg tcgctaagcg aaagctaagc aaataaacaa gcgcagctga
     4381 acaagctaaa caatcggggt accgctagag tcgacggtac gatccaccgg tcgccaccat
     4441 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     4501 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     4561 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     4621 cgtgaccacc ctgacctggg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     4681 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     4741 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     4801 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     4861 gctggagtac aactacatca gccacaacgt ctatatcacc gccgacaagc agaagaacgg
     4921 catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     4981 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     5041 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     5101 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaag
     5161 cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta
     5221 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     5281 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     5341 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     5401 taaagcttct aggccggccg ggccctcatt catcttagtt atatactcaa tggaaacggt
     5461 aaaagttcta gcgatgggcg aaaatatttt ttcctatcca taaagagagc tttaagttag
     5521 tcctctttga cctttttatt ataatatttg atttgacctc gttggctttc aatgacattc
     5581 tttgagagaa caatgagtat cttaaacgct caaataaata ttacctatcg catctcttca
     5641 tagtaaaata aatgttccaa acagatccgt gtttgcgatt gcatatttca aggcgtaatg
     5701 tatttttcgg aggatggatg ctgacaatga aaccgtatcc gttatcacca tggagaggtg
     5761 ttgaagaacg aacaattaag aagaaagttt tttgttttcg gcatttgata ggatcttacc
     5821 actcgaagcc aatccagcga gacaaatagc ttgaagttat ttagcgcatc gttaaatttt
     5881 taacgactct aattggagct gccagacaaa tttgatttgt ccgtgcggta tgcatatgta
     5941 cctacttacg tctgtcgttt tgtctccgtt taccgggagg gaaagtctgg aaacatggaa
     6001 actctatagt tgccaggtag accatctgcc tccgtcggct ggctggattc caatttgaat
     6061 attggctaat tggaagagat ggaagttttt gaatggatga ttgaataatt gaagcgactc
     6121 cgggtacctg tttgtaagct ctgcaacagt gccatagatt cgtgtcagtc catcactaga
     6181 atcaaatcaa cttgtacttg atatataaat ggcttgggtt aatggtgcga tcttgacccc
     6241 cgggtcttag agctatgctg gaaacagcat agcaagttaa gataaggcta gtccgttatc
     6301 aacttgaaaa agtggcaccg agtcggtgct tttttggcgc gccgtcgacg atgtaggtca
     6361 cggtctcgaa gccgcggtgc gggtgccagg gcgtgccctt gggctccccg ggcgcgtact
     6421 ccacctcacc catctggtcc atcatgatga acgggtcgag gtggcggtag ttgatcccgg
     6481 cgaacgcgcg gcgcaccggg aagccctcgc cctcgaaacc gctgggcgcg gtggtcacgg
     6541 tgagcacggg acgtgcgacg gcgtcggcgg gtgcggatac gcggggcagc gtcagcgggt
     6601 tctcgacggt cacggcgggc atgtcgacgg ccggccgatc tcggatctga caatgttcag
     6661 tgcagagact cggctacgcc tcgtggactt tgaagttgac caacaatgtt tattcttacc
     6721 tctaatagtc ctctgtggca aggtcaagat tctgttagaa gccaatgaag aacctggttg
     6781 ttcaataaca ttttgttcgt ctaatatttc actaccgctt gacgttggct gcacttcatg
     6841 tacctcatct ataaacgctt cttctgtatc gctctggacg tcatcttcac ttacgtgatc
     6901 tgatatttca ctgtcagaat cctcaccaac aagctcgtca tcgctttgca gaagagcaga
     6961 gaggatatgc tcatcgtcta aagaactacc cattttatta tatattagtc acgatatcta
     7021 taacaagaaa atatatatat aataagttat cacgtaagta gaacatgaaa taacaatata
     7081 attatcgtat gagttaaatc ttaaaagtca cgtaaaagat aatcatgcgt cattttgact
     7141 cacgcggtcg ttatagttca aaatcagtga cacttaccgc attgacaagc acgcctcacg
     7201 ggagctccaa gcggcgactg agatgtccta aatgcacagc gacggattcg cgctatttag
     7261 aaagagagag caatatttca agaatgcatg cgtcaatttt acgcagacta tctttctagg
     7321 gttaaaaaag atttgcgctt tactcgacct aaactttaaa cacgtcatag aatcttcgtt
     7381 tgacaaaaac cacattgtgg ccaagctgtg tgacgcgacg cgcgctaaag aatggcaaac
     7441 caagtcgcgc gagcgtcgac tctagaggat ccccgggtac cgagctcgaa ttcgtaatca
     7501 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     7561 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     7621 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     7681 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     7741 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     7801 gtaatacggt
//
