LOCUS       sequence-249750-        7847 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 7847)
  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 7847)
  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: U6c-T
FEATURES             Location/Qualifiers
     source          1..7847
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..3,4..6,7..720)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(46..67)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(307..326)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     654..675
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    843..964
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(880..899)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     934..953
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    1957..2026
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     repeat_region   2879..2913
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(3094..3110)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3094..3110)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3107..3129)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3118..3134
                     /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(3142..3148,3149..3166,3167..3172))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3187..3208
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3325..3342)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3496..4084)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3576..3595)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4255..5046,5047..5115))
                     /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     4878..4897
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5116..5220)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5288..5306
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5344..5366)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5466..5485
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5679..5701
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5693..5710
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5694..5710
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   6414..6476
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     regulatory      join(7842..7847,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
       61 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      121 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      181 ctcgtgacca ccctgacctg gggcgtgcag tgcttcagcc gctaccccga ccacatgaag
      241 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      301 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      361 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      421 aagctggagt acaactacat cagccacaac gtctatatca ccgccgacaa gcagaagaac
      481 ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
      541 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      601 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
      661 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
      721 agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
      781 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
      841 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
      901 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
      961 cttaaagctt ctaggccggc cgggcccata tgtcttgccg aagcacatta acccaagcca
     1021 tttatatatc aagtacaagt tgatttgatt ctagtgatgg actgacacga atctatggca
     1081 ctgttgcaga gcttacaaac aggtacccgg agtcgcttca attattcaat catccattca
     1141 aaaacttcca tctcttccaa ttagccaata ttcaaattgg aatccagcca gccgacggag
     1201 gcagatggtc tacctggcaa ctatagagtt tccatgtttc cagactttcc ctcccggtaa
     1261 acggagacaa aacgacagac gtaagtaggt acatatgcat accgcacgga caaatcaaat
     1321 ttgtctggca gctccaatta gagtcgttaa aaatttaacg atgcgctaaa taacttcaag
     1381 ctatttgtct cgctggattg gcttcgagtg gtaagatcct atcaaatgcc gaaaacaaaa
     1441 aactttcttc ttaattgttc gttcttcaac acctctccat ggtgataacg gatacggttt
     1501 cattgtcagc atccatcctc cgaaaaatac attacgcctt gaaatatgca atcgcaaaca
     1561 cggatctgtt tggaacattt attttactat gaagagatgc gataggtaat atttatttga
     1621 gcgtttaaga tactcattgt tctctcaaag aatgtcattg aaagccaacg aggtcaaatc
     1681 aaatattata ataaaaaggt caaagaggac taacttaaag ctctctttat ggataggaaa
     1741 aaatattttc gcccatcgct agaactttta ccgtttccat tgagtatata actaagatga
     1801 atgaggctaa ttgatggtgc gatcttgacc cccgggtctt agagctatgc tggaaacagc
     1861 atagcaagtt aagataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
     1921 cttttttggc gcgccgtcga cgatgtaggt cacggtctcg aagccgcggt gcgggtgcca
     1981 gggcgtgccc ttgggctccc cgggcgcgta ctccacctca cccatctggt ccatcatgat
     2041 gaacgggtcg aggtggcggt agttgatccc ggcgaacgcg cggcgcaccg ggaagccctc
     2101 gccctcgaaa ccgctgggcg cggtggtcac ggtgagcacg ggacgtgcga cggcgtcggc
     2161 gggtgcggat acgcggggca gcgtcagcgg gttctcgacg gtcacggcgg gcatgtcgac
     2221 ggccggccga tctcggatct gacaatgttc agtgcagaga ctcggctacg cctcgtggac
     2281 tttgaagttg accaacaatg tttattctta cctctaatag tcctctgtgg caaggtcaag
     2341 attctgttag aagccaatga agaacctggt tgttcaataa cattttgttc gtctaatatt
     2401 tcactaccgc ttgacgttgg ctgcacttca tgtacctcat ctataaacgc ttcttctgta
     2461 tcgctctgga cgtcatcttc acttacgtga tctgatattt cactgtcaga atcctcacca
     2521 acaagctcgt catcgctttg cagaagagca gagaggatat gctcatcgtc taaagaacta
     2581 cccattttat tatatattag tcacgatatc tataacaaga aaatatatat ataataagtt
     2641 atcacgtaag tagaacatga aataacaata taattatcgt atgagttaaa tcttaaaagt
     2701 cacgtaaaag ataatcatgc gtcattttga ctcacgcggt cgttatagtt caaaatcagt
     2761 gacacttacc gcattgacaa gcacgcctca cgggagctcc aagcggcgac tgagatgtcc
     2821 taaatgcaca gcgacggatt cgcgctattt agaaagagag agcaatattt caagaatgca
     2881 tgcgtcaatt ttacgcagac tatctttcta gggttaaaaa agatttgcgc tttactcgac
     2941 ctaaacttta aacacgtcat agaatcttcg tttgacaaaa accacattgt ggccaagctg
     3001 tgtgacgcga cgcgcgctaa agaatggcaa accaagtcgc gcgagcgtcg actctagagg
     3061 atccccgggt accgagctcg aattcgtaat catgtcatag ctgtttcctg tgtgaaattg
     3121 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     3181 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3241 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3301 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3361 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3421 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3481 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3541 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3601 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3661 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3721 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3781 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3841 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3901 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3961 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4021 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4081 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4141 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4201 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4261 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4321 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4381 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4441 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4501 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4561 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4621 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4681 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4741 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4801 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4861 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4921 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4981 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5041 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5101 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5161 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5221 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     5281 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     5341 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     5401 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     5461 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     5521 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     5581 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     5641 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     5701 gacggccagt gccaagcttt gtttaaaaat ataacaaaat tgtgatccca caaaatgaag
     5761 tggggcaaaa tcaaataatt aatagtgtcc gtaaacttgt tggtcttcaa ctttttgagg
     5821 aacacgttgg acggcaaatc cgtgactata acacaagttg atttaataat tttagccaac
     5881 acgtcgggct gcgtgttttt tgccgacgcg tctgtgtaca cgttgattaa ctggtcgatt
     5941 aaactgttga aataatttaa tttttggttc ttctttaaat ctgtgatgaa attttttaaa
     6001 ataactttaa attcttcatt ggtaaaaaat gccacgtttt gcaacttgtg agggtctaat
     6061 atgaggtcaa actcagtagg agttttatcc aaaaaagaaa acatgattac gtctgtacac
     6121 gaacgcgtat taacgcagag tgcaaagtat aagagggtta aaaaatatat tttacgcacc
     6181 atatacgcat cgggttgata tcgttaatat ggatcaattt gaacagttga ttaacgtgtc
     6241 tctgctcaag tctttgatca aaacgcaaat cgacgaaaat gtgtcggaca atatcaagtc
     6301 gatgagcgaa aaactaaaaa ggctagaata cgacaatctc acagacagcg ttgagatata
     6361 cggtattcac gacagcaggc tgaataataa aaaaattaga aactattatt taaccctaga
     6421 aagataatca tattgtgacg tacgttaaag ataatcatgc gtaaaattga cgcatgtgtt
     6481 ttatcggtct gtatatcgag gtttatttat taatttgaat agatattaag ttttattata
     6541 tttacactta catactaata ataaattcaa caaacaattt atttatgttt atttatttat
     6601 taaaaaaaaa caaaaactca aaatttcttc tataaagtaa caaaactttt aaacattctc
     6661 tcttttacaa aaataaactt attttgtact ttaaaaacag tcatgttgta ttataaaata
     6721 agtaattagc ttaacttata cataatagaa acaaattata cttattagtc agtcagaaac
     6781 aactttggca catatcaata ttatgctctc gacaaataac ttttttgcat tttttgcacg
     6841 atgcatttgc ctttcgcctt attttagagg ggcagtaagt acagtaagta cgttttttca
     6901 ttactggctc ttcagtactg tcatctgatg taccaggcac ttcatttggc aaaatattag
     6961 agatattatc gcgcaaatat ctcttcaaag taggagcttc taaacgctta cgcataaacg
     7021 atgacgtcag gctcatgtaa aggtttctca taaatttttt gcgactttga accttttctc
     7081 ccttgctact gacattatgg ctgtatataa taaaagaatt tatgcaggca atgtttatca
     7141 ttccgtacaa taatgccata ggccacctat tcgtcttcct actgcaggtc atcacagaac
     7201 acatttggtc tagcgtgtcc actccgcctt tagtttgatt ataatacata accatttgcg
     7261 gtttaccggt actttcgttg atagaagcat cctcatcaca agatgataat aagtatacca
     7321 tcttagctgg cttcggttta tatgagacga gagtaagggg tccgtcaaaa caaaacatcg
     7381 atgttcccac tggcctggag cgactgtttt tcagtacttc cggtatctcg cgtttgtttg
     7441 atcgcacggt tcccacaatg gttaattcga gctcgcccgg gcttaagagt tcccacaatg
     7501 gttaattcga gctcgcccgg gcttaagagt tcccacaatg gttaattcga gctcgcccgg
     7561 gcttaagagt tcccacaatg gttaattcga gctcgcccgg ggatctaatt caattagaga
     7621 ctaattcaat tagagctaat tcaattagga tccaagctta tcgatttcga accctcgacc
     7681 gccggagtat aaatagaggc gcttcgtcta cggagcgaca attcaattca aacaagcaaa
     7741 gtgaacacgt cgctaagcga aagctaagca aataaacaag cgcagctgaa caagctaaac
     7801 aatcggggta ccgctagagt cgacggtacg atccaccggt cgccacc
//
