LOCUS       sequence-82256-e        7728 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Left hand TALEN against Aedes aegypti KMO gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7728)
  AUTHORS   Aryan A, Anderson MA, Myles KM, Adelman ZN
  TITLE     TALEN-based gene disruption in the dengue vector Aedes aegypti.
  JOURNAL   PLoS One. 2013;8(3):e60082. doi: 10.1371/journal.pone.0060082. Epub 
            2013 Mar 21.
  PUBMED    23555893
REFERENCE   2  (bases 1 to 7728)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2013"; volume: "8(3):e60082. doi"; pages: " 
            10.1371/journal.pone.0060082. Epub 2013 Mar 21"
COMMENT     Sequence Label: pSLfa PUb TAL-KMO-L
FEATURES             Location/Qualifiers
     source          1..7728
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(12..29)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(183..771)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(263..282)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(942..1733,1734..1802))
                     /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     1565..1584
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1803..1907)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1975..1993
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2031..2053)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2153..2172
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2189..2644)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2326..2347)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2538..2557
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2842..2864
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2856..2873
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2857..2873
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4377..4397
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4404..4430
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6579..7166
                     /codon_start=1
                     /product="nonspecific DNA cleavage domain of the FokI 
                     endonuclease (Li et al., 1992)"
                     /label=FokI cleavage domain
                     /note="must dimerize to be catalytically active"
                     /translation="QLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKV
                     MEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQR
                     YVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVL
                     SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
     polyA_signal    7351..7432
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7388..7407)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7442..7461
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7509..7525)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7509..7525)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7522..7544)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7533..7549
                     /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(7557..7563,7564..7581,7582..7587))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7602..7623
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
       61 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
      121 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
      181 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      241 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      301 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      361 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      421 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      481 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      541 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      601 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
      661 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      721 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      781 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
      841 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
      901 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
      961 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     1021 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     1081 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     1141 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     1201 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     1261 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     1321 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     1381 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     1441 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     1501 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     1561 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     1621 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     1681 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     1741 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     1801 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     1861 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     1921 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     1981 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     2041 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     2101 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     2161 gagcagattg tactgagagt gcaccataaa attgtaaacg ttaatatttt gttaaaattc
     2221 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     2281 ccttataaat caaaagaata gcccgagata gggttgagtg ttgttccagt ttggaacaag
     2341 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     2401 gatggcccac tacgtgaacc atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa
     2461 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     2521 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     2581 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     2641 gcgtactatg gttgctttga cgtatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     2701 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     2761 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     2821 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc
     2881 tggccggcct aggcgcgcca agcttaaggt gcacggccca cgtggccact agtacttctc
     2941 gagctctgta catgtccgcg gtcgcgacgt acgcgtatct ttacatgtag cttgtgcatt
     3001 gaatccaatt ataatttgcc ttggcaccag ctgagccaga caagaaagaa agcttcccag
     3061 aagtatatcg atttagaagg gttgacgtca cttgctgact gcactaatac agcaaatgat
     3121 gcaattagaa tgattcaagt gaaattccca aattactgat ttttctctgg atttggttat
     3181 cagattacat tcgaagctaa gattagctac cgaaattgtc gatcaaatca ggaaatcctt
     3241 tctctatcga aaaaggcatt cgcacatctt cctctctatg ccatatacac gaagggtagg
     3301 tacattgacg tctttgccag aagttgaact gcatcgttca aggtacagaa tgaacgacta
     3361 acagacacaa gcacgttttg ctgtccattc agacacaggg atggtaccca tattcgatcg
     3421 atatagagcc atccaaccga acagaggtat atgtatgaat gtattgctga aattttctag
     3481 aagtacaacc accactacga cagtgtctat aaaacgcccc tgcaaaggcg aaaccagctc
     3541 aatcgaatac gtttcctagt ggagtgaaca ttacgcggcc caagtaagca gtgccagtgc
     3601 aagtgaagtg aagtctctag tgaaaaagag tgatccaatt agccagagga gaaaatttca
     3661 gagtgaacaa agctttattc aaaggacaat tactattaaa ttggtgaaag tgcatttcgg
     3721 tgaagggaat cttctagtga aggtaggtaa attaattgat gaaattatag ctatgagcga
     3781 aaactagttt ggtgaatgat tcctttgtct ttgaatgagc aaactatttt ccaagatggc
     3841 gactattgag ctttgagtga ttagtgaaaa tttgcaacgc agtttcatca tcattgataa
     3901 aacccaattg tgattcacag cgataatcat atttcgttga atcatcgcta ctaattgaat
     3961 taaatttcta gaataataag aataacgtat ttgctccgtc acatatctaa aataaatatt
     4021 ttgatggtaa ttacccatta aggtaatatt aacacatatc gagaaaaacc ttgaggaaat
     4081 cgtgaaaact tgaagatacg caatttcaaa actacgtagt tcaaagtcga aaacaagtta
     4141 atttttcact taaaagtagg gcgttgttgt gacgtcatca ccttcaagtg tatatttttc
     4201 acttggcctg cgactgcaaa cgcagacaaa gcaaaacaag tttaaaacct gtcgtgtcgt
     4261 gctcgaagcc aaaggcaatg aatcaatatc aaatgagagt ttgcatttca caaccaatta
     4321 ctcaagcgtt tcctcgtttc tttttctgct caacagagat ttcaaccatg ggcgatccta
     4381 aaaagaaacg taaggtcatc gattacccat acgatgttcc agattacgct atcgatatcg
     4441 ccgatctacg cacgctcggc tacagccagc agcaacagga gaagatcaaa ccgaaggttc
     4501 gttcgacagt ggcgcagcac cacgaggcac tggtcggcca cgggtttaca cacgcgcaca
     4561 tcgttgcgtt aagccaacac ccggcagcgt tagggaccgt cgctgtcaag tatcaggaca
     4621 tgatcgcagc gttgccagag gcgacacacg aagcgatcgt tggcgtcggc aaacagtggt
     4681 ccggcgcacg cgctctggag gccttgctca cggtggcggg agagttgaga ggtccaccgt
     4741 tacagttgga cacaggccaa cttctcaaga ttgcaaaacg tggcggcgtg accgcagtgg
     4801 aggcagtgca tgcatggcgc aatgcactga cgggtgcccc gctcaacttg accccccagc
     4861 aggtggtggc catcgccagc aataatggtg gcaagcaggc gctggagacg gtccagcggc
     4921 tgttgccggt gctgtgccag gcccacggct tgaccccgga gcaggtggtg gccatcgcca
     4981 gcaatattgg tggcaagcag gcgctggaga cggtgcaggc gctgttgccg gtgctgtgcc
     5041 aggcccacgg cttgaccccg gagcaggtgg tggccatcgc cagcaatatt ggtggcaagc
     5101 aggcgctgga gacggtgcag gcgctgttgc cggtgctgtg ccaggcccac ggcttgaccc
     5161 cccagcaggt ggtggccatc gccagcaatg gcggtggcaa gcaggcgctg gagacggtcc
     5221 agcggctgtt gccggtgctg tgccaggccc acggcttgac cccccagcag gtggtggcca
     5281 tcgccagcaa taatggtggc aagcaggcgc tggagacggt ccagcggctg ttgccggtgc
     5341 tgtgccaggc ccacggcttg accccggagc aggtggtggc catcgccagc cacgatggcg
     5401 gcaagcaggc gctggagacg gtccagcggc tgttgccggt gctgtgccag gcccacggct
     5461 tgaccccgga gcaggtggtg gccatcgcca gccacgatgg cggcaagcag gcgctggaga
     5521 cggtccagcg gctgttgccg gtgctgtgcc aggcccacgg cttgaccccc cagcaggtgg
     5581 tggccatcgc cagcaataat ggtggcaagc aggcgctgga gacggtccag cggctgttgc
     5641 cggtgctgtg ccaggcccac ggcttgaccc cccagcaggt ggtggccatc gccagcaata
     5701 atggtggcaa gcaggcgctg gagacggtcc agcggctgtt gccggtgctg tgccaggccc
     5761 acggcttgac cccggagcag gtggtggcca tcgccagcaa tattggtggc aagcaggcgc
     5821 tggagacggt gcaggcgctg ttgccggtgc tgtgccaggc ccacggcttg accccccagc
     5881 aggtggtggc catcgccagc aatggcggtg gcaagcaggc gctggagacg gtccagcggc
     5941 tgttgccggt gctgtgccag gcccacggct tgacccccca gcaggtggtg gccatcgcca
     6001 gcaatggcgg tggcaagcag gcgctggaga cggtccagcg gctgttgccg gtgctgtgcc
     6061 aggcccacgg cttgaccccg gagcaggtgg tggccatcgc cagccacgat ggcggcaagc
     6121 aggcgctgga gacggtccag cggctgttgc cggtgctgtg ccaggcccac ggcttgaccc
     6181 cccagcaggt ggtggccatc gccagcaata atggtggcaa gcaggcgctg gagacggtcc
     6241 agcggctgtt gccggtgctg tgccaggccc acggcttgac cccggagcag gtggtggcca
     6301 tcgccagcaa tattggtggc aagcaggcgc tggagacggt gcaggcgctg ttgccggtgc
     6361 tgtgccaggc ccacggcttg acccctcagc aggtggtggc catcgccagc aatggcggcg
     6421 gcaggccggc gctggagagc attgttgccc agttatctcg ccctgatccg gcgttggccg
     6481 cgttgaccaa cgaccacctc gtcgccttgg cctgcctcgg cgggcgtcct gcgctggatg
     6541 cagtgaaaaa gggattgggg gatcctatca gccgttccca gctggtgaag tccgagctgg
     6601 aggagaagaa atccgagttg aggcacaagc tgaagtacgt gccccacgag tacatcgagc
     6661 tgatcgagat cgcccggaac agcacccagg accgtatcct ggagatgaag gtgatggagt
     6721 tcttcatgaa ggtgtacggc tacaggggca agcacctggg cggctccagg aagcccgacg
     6781 gcgccatcta caccgtgggc tcccccatcg actacggcgt gatcgtggac accaaggcct
     6841 actccggcgg ctacaacctg cccatcggcc aggccgacga aatgcagagg tacgtggagg
     6901 agaaccagac caggaacaag cacatcaacc ccaacgagtg gtggaaggtg tacccctcca
     6961 gcgtgaccga gttcaagttc ctgttcgtgt ccggccactt caagggcaac tacaaggccc
     7021 agctgaccag gctgaaccac atcaccaact gcaacggcgc cgtgctgtcc gtggaggagc
     7081 tcctgatcgg cggcgagatg atcaaggccg gcaccctgac cctggaggag gtgaggagga
     7141 agttcaacaa cggcgagatc aacttcgcgg ccgactgata actcgagcga tcctctagac
     7201 gagctcctcg agcctgcaga tctgtcgacg cggccgcgac tctagatcat aatcagccat
     7261 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     7321 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     7381 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     7441 tgtggtttgt ccaaactcat caatgtatct tgaattcggc cggcctaggc gcgccaattc
     7501 gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     7561 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     7621 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     7681 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgc
//
