LOCUS Exported 9463 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Anopheles docking site transgenesis plasmid expressing Cre recombinase and I-SceI nuclease, vasa promoter, DsRed marker. ACCESSION . VERSION . KEYWORDS pDSAR-Cre-F2A-I-SceI SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9463) AUTHORS Volohonsky G, Terenzi O, Soichot J, Naujoks DA, Nolan T, Windbichler N, Kapps D, Smidler AL, Vittu A, Costa G, Steinert S, Levashina EA, Blandin SA, Marois E TITLE Tools for Anopheles gambiae Transgenesis. JOURNAL G3 (Bethesda). 2015 Apr 13. pii: g3.115.016808. doi: 10.1534/g3.115.016808. PUBMED 25869647 REFERENCE 2 (bases 1 to 9463) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9463 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 92..109 /label=L4440 /note="L4440 vector, forward primer" protein_bind complement(259..279) /gene="mutant version of attB" /label=attB3 /bound_moiety="BP Clonase(TM)" /note="core recombination site for the Gateway(R) BP reaction" CDS complement(1996..2022) /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS complement(2041..2061) /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS complement(2113..2178) /codon_start=1 /product="2A peptide from foot-and-mouth disease virus polyprotein" /label=F2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="VKQTLNFDLLKLAGDVESNPGP" CDS complement(2197..3225) /codon_start=1 /product="site-specific recombinase" /label=Cre /note="Cre is a site-specific recombinase from bacteriophage P1. Recombination occurs at loxP sequences." /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE DGD" CDS complement(3244..3264) /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS complement(3319..3345) /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS complement(3364..3384) /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 5862..5944 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" protein_bind 6002..6071 /label=attB /bound_moiety="phage phi-C31 integrase" /note="attB site for the phi-C31 integrase (Groth et al., 2000)" regulatory 6586..6595 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 6592..7272 /codon_start=1 /product="wild-type DsRed" /label=DsRed1 /note="mammalian codon-optimized" /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH HLFL" primer_bind complement(6775..6795) /label=DsRed1-N /note="DsRed1, reverse primer" primer_bind 6993..7012 /label=mCherry-F /note="mCherry, forward primer" primer_bind 7187..7210 /label=DsRed1-C /note="DsRed1, forward primer" polyA_signal 7393..7514 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" protein_bind complement(7575..7674) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(7704..7723) /label=T7 /note="T7 promoter, forward primer" promoter complement(7705..7723) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(7728..7744) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7728..7744) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7846..7865) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 7857..8666 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(7924..7943) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 8813..9401 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 9302..9321 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga 61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga 121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca 181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc 241 tagcgggggg cccggtacca actttattat acaaagttgt ctcgagaaaa tgtggccatt 301 aacagcagtt ttttttaaaa cgatgtatca accatctaac atgcatattt tacaattttc 361 aggaaaataa tcaagctcta atattgtact aaagaaacat atagatatag taataatact 421 atagatatag aaatttcaaa tattctcctt tttggaatag ttagacggtc atggtggcct 481 atactggctt ttggaattta ctgagtttac gccgccagtt acaagggatc ggtccatttg 541 aggtaagaac cctcgatggg catgttgtca agtctagcag tatgacgttt gtaccatggt 601 accgctccgt aaattgaata tgttattagt attttcttag ccaaacgaga cataaacgat 661 aagccactag ataaaggata catcaagatg gtgctctaag cggcccgcgg gctgataata 721 aagtttgaag acccctgaac gatatgttga caatgaacgg gctttgtgca gttacgctca 781 attcgtcgtt atcttttttg aaacctgggt tagtctaagt actttgtaca actaccttca 841 cttgtgtatt ttatcctgga atggacctaa aggtgacgta ttgttagaat acttcagttc 901 tagagtatac aagtgcgaaa aatcaccttt cagtacatta cattgaataa tgaaagacaa 961 tcggtgcaag ctttcacgaa ttatagaata aaagtgcaag ttcttatgtt ttcaaagttt 1021 tcatcaacgt tttattaacg aacaattctt tgctgtgtgt catgcttctt ctgtgtgttt 1081 tacatcgtta cgtcagtaaa tgaaagaata atcatgcatg tcacaaaagc aaggaaagga 1141 gtttgactcc tcgtttcgaa tgaggaaact tctcttccga agtttcgttg taagtgttaa 1201 caatagaaaa aaaaaataag gaaatgtcga gagagatcca ctctcgttcg caacacataa 1261 caaccccacc ccaagctcga gctcttattt caggaaagtt tcggaggaga tagtgttcgg 1321 cagtttgtac atcatctgcg ggatcaggta cggtttgatc aggttgtaga agatcaggta 1381 agacatagaa tcgatgtaga tgatcggttt gtttttgttg atttttacgt aacagttcag 1441 ttggaatttg ttacgcagac ccttaaccag gtattctact tcttcgaaag tgaaagactg 1501 ggtgttcagt acgatcgatt tgttggtaga gtttttgttg taatcccatt taccaccatc 1561 atccatgaac cagtatgcca gagacatcgg ggtcaggtag ttttcaacca ggttgttcgg 1621 gatggttttt ttgttgttaa cgatgaacag gttagccagt ttgttgaaag cttggtgttt 1681 gaaagtctgg gcgccccagg tgattaccag gttacccagg tggttaacac gttctttttt 1741 gtgcggcggg gacagtaccc actgatcgta cagcagacat acgtggtcca tgtatgcttt 1801 gtttttccac tcgaactgca tacagtaggt tttaccttca tcacgagaac ggatgtaagc 1861 atcacccagg atcagaccga tacctgcttc gaactgttcg atgttcagtt cgatcagctg 1921 ggatttgtat tctttcagca gtttagagtt cggacccagg ttcattacct ggtttttttt 1981 gatgtttttc atacccgcat agtcaggaac atcgtatggg tacataaatc gtgggtcttc 2041 tacttttcgt tttttttttg gtggtgctgc atgctgcgca tcggccgtag cttcgtcgtc 2101 tgatgatgat cctggtccgg gattcgactc cacgtctcca gccagcttca gcaggtcgaa 2161 gttcagggtc tgcttcaccg gagcacgctt ggcgcgatcg ccatcttcca gcaggcgcac 2221 cattgcccct gtttcactat ccaggttacg gatatagttc atgacaatat ttacattggt 2281 ccagccacca gcttgcatga tctccggtat tgaaactcca gcgcgggcca tatctcgcgc 2341 ggctccgaca cgggcactgt gtccagacca ggccaggtat ctctgaccag agtcatcctt 2401 agcgccgtaa atcaatcgat gagttgcttc aaaaatccct tccagggcgc gagttgatag 2461 ctggctggtg gcagatggcg cggcaacacc attttttctg acccggcaaa acaggtagtt 2521 attcggatca tcagctacac cagagacgga aatccatcgc tcgaccagtt tagttacccc 2581 caggctaagt gccttctcta cacctgcggt gctaaccagc gttttcgttc tgccaatatg 2641 gattaacatt ctcccaccgt cagtacgtga gatatcttta accctgatcc tggcaatttc 2701 ggctatacgt aacagggtgt tataagcaat ccccagaaat gccagattac gtatatcctg 2761 gcagcgatcg ctattttcca tgagtgaacg aacctggtcg aaatcagtgc gttcgaacgc 2821 tagagcctgt tttgcacgtt caccggcatc aacgttttct tttcggatcc gccgcataac 2881 cagtgaaaca gcattgctgt cacttggtcg tggcagcccg gaccgacgat gaagcatgtt 2941 tagctggccc aaatgttgct ggatagtttt tactgccaga ccgcgcgcct gaagatatag 3001 aagataatcg cgaacatctt caggttctgc gggaaaccat ttccggttat tcaacttgca 3061 ccatgccgcc cacgaccggc aaacggacag aagcattttc caggtatgct cagaaaacgc 3121 ctggcgatcc ctgaacatgt ccatcaggtt cttgcgaacc tcatcactcg ttgcatcgac 3181 cggtaatgca ggcaaatttt ggtgtacggt cagtaaattg gacatgcggc cgcccgggtc 3241 ttctaccttt ctcttctttt ttggagggcc gctcatgaaa gggcgaattc cctggttttt 3301 tttgatgttt ttcatacccg catagtcagg aacatcgtat gggtacataa atcgtgggtc 3361 ttctactttt cgtttttttt ttggtggtgc tgcatgctgc gcatcggccg tagcttcgtc 3421 gtctgatgat gatcccatgt ttccggatcc gtttcctttc tttattcacc ggaagatgac 3481 aattctcctt atgggaatgt acgattcgag atctgtgtgt ctcccgaaga tgaaaaaagc 3541 tgttggtttc aaccaaaaaa cgtgtccgtt gctcgaagcg cgactaacgg cgactctctt 3601 tcgactagca atggcgtccg caggcacaaa ccgagaggaa tagtggaaca tgcaatgcag 3661 cagcagcagc aacagcagca gcacgtggag agagagaaag aaagatggtc gaaatattac 3721 aaaaaataat tgccgttgca cacacacaca cacacacaca cacacacaca cacacacaca 3781 cacacacaca cacacacaca cacacacaca cacacacaca cacacacaca cactacacca 3841 ccaatatcga cgaccaccgc tctacccgca cccattgcaa cctaaagaga cgatggtatt 3901 ttccagcgaa attttcattg tccattgcaa gcgacctcac acaccgccac cgaagtagcc 3961 tttgaagcac gcgcgcgtgt gaaggcgtgt gcctctgtgt gtgtaggtga atatcactgt 4021 gtacactgcg ctaagccagt ggcgcgcgcg cttttcgcat caattattgc cggatgaacg 4081 acaacgagat gctcttcgcg gggcccgctg tcccggaacg tgcacgaaag caccatttct 4141 cctctcggaa actccacatc tcggaaaggc ttcccaacta tgccccaacg actctcttat 4201 ctattgcata tcgaagactt ttgaatccgc tccagtgcca actgcggttc atgaaacacg 4261 aaactgtttt tctcccctga ttcgagcaaa caagagcacc atggtaagct gtggaaataa 4321 cgccatcctt ctggatatct tgtaacgaaa aaggccaagg actgtgattc tactaacctc 4381 actctggaac tgatatgaga gcacacagag ccacacaacc actccacgac gggatacgct 4441 tggattaaaa ctaaaccgct tccgtacgtc gacaccgact ggcccctata ccgcttctca 4501 aataacgacg atctcagcta aaatgccccc cacacacatg catgcgcagc acacttgttc 4561 acttccactt ccttgcctgc tctgtgtgcg gtggtggata cgtgctgtgc tgtaagaaca 4621 cttcacacag aaaggaaggg acagagcaaa ccaacacact gccaccgctc ttctcgtgcc 4681 atttccttag aaaattaatc caaaaactca actgcgccac acaccacggc agccacggca 4741 gcctgctatc aaaagggggg aggcgcgcaa catgcacaac acacgataca caccacggaa 4801 tgtttacgaa cagcgtgcga acaggcaatt tcggtaccgg cggcaacagc cgattttgac 4861 agagcgctca acacacacac gcacacactc taataccagc aacgcgtgcg aaggcttttg 4921 tttcgcactg ccagtgtgtt agtgggagca ggtacaaaat gaaacaaagg gcgaatggcg 4981 cgtgctactg aacaatacaa gcgaattcga gaaaaattaa ataagccaag agattatttt 5041 atgagtaaat gaaatgaaat ttaatttaac tgtattttga acataactaa gaaaatacta 5101 atcaactgga tgaaataata taacatcaca aaacaataat tcaatttgaa atcagttacg 5161 ttagttttgc cgaaatgaga gatttttttc acaaacactt cactaacttc taacaaacaa 5221 acatttgttc tataaaacta gttcacttac ctgagttttg aagtactatt cggctggtta 5281 tttcacttgc aaaagcaatt ggctaattta gcgctaagca cctcagcaga aacctccgac 5341 tcgccgcgag tgctccgagc aaaagactgc taaaagctgg aatttgcacg gacacggcct 5401 actcgtccgt cgtactgtgc aactaggatg gaagatttcg atacgggact ctacagggag 5461 ggtttatatg tgttttataa tgctgcatgt atgtttttat gtataaaata tgcaaaaagt 5521 atgtcttttt agtttattat aacaaataaa atacaataca aattataaaa caatcaattt 5581 ctcaacggtt ttgtttgcag cgagttgtga tttgcttgaa aacaaagcgc acgctgtaat 5641 tttcccgccc agttcggcat gtcaaacacg tctgacgttc ggcttcccac tgtagctgct 5701 gctgtcatgg aaggaaaaga atttgctcgc gttctacatc gataccgtcg acgctagaca 5761 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc 5821 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt 5881 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttcttca 5941 ctgcattcta gttgtggttt gtccaaactc atcaatgtat ctcgacgatg taggtcacag 6001 tctcgaagcc gcggtgcggg tgccagggcg tgcccttggg ctccccgggc gcgtactcca 6061 cctcacccat ctggtccatc atgatgaacg ggtcgaggtg gcggtagttg atcccggcga 6121 acgcgcggcg caccgggaag ccctcgccct cgaaaccgct gggcgcggtg gtcacggtga 6181 gcacgggacg tgcgacggcg tcggcgggtg cggatacgcg gggcagcgtc agcgggttct 6241 cgacggtcac ggcgggcaat tcctgcagac ttccggtatc tcgcgtttgt ttgatcgcac 6301 ggttcccaca atggttaatt cgagctcgcc cggggatcta attcaattag agactaattc 6361 aattagagct aattcaatta ggatccaagc ttatcgattt cgaaccctcg accgccggag 6421 tataaataga ggcgcttcgt ctacggagcg acaattcaat tcaaacaagc aaagtgaaca 6481 cgtcgctaag cgaaagctaa gcaaataaac aagcgcagct gaacaagcta aacaatcggg 6541 gtaccgctag agtcgacggt accgcgggcc cgggatccac cggtcgccac catggtgcgc 6601 tcctccaaga acgtcatcaa ggagttcatg cgcttcaagg tgcgcatgga gggcaccgtg 6661 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggccacaac 6721 accgtgaagc tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc 6781 ccccagttcc agtacggctc caaggtgtac gtgaagcacc ccgccgacat ccccgactac 6841 aagaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc 6901 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gctgcttcat ctacaaggtg 6961 aagttcatcg gcgtgaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc 7021 tgggaggcct ccaccgagcg cctgtacccc cgcgacggcg tgctgaaggg cgagatccac 7081 aaggccctga agctgaagga cggcggccac tacctggtgg agttcaagtc catctacatg 7141 gccaagaagc ccgtgcagct gcccggctac tactacgtgg actccaagct ggacatcacc 7201 tcccacaacg aggactacac catcgtggag cagtacgagc gcaccgaggg ccgccaccac 7261 ctgttcctgt agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt 7321 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca 7381 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 7441 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 7501 atcaatgtat cttaaagctt atcgatacgc gtacggcgcg cctagagcgg ccgccaccgc 7561 ggtggagctc gagtacccag ctttcttgta caaagttggc attataagaa agcattgctt 7621 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc 7681 agctgcaggg cggccgcgat atcccctata gtgagtcgta ttacatggtc atagctgttt 7741 cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata 7801 tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga 7861 gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat 7921 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt 7981 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg 8041 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca 8101 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atccccggaa 8161 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc 8221 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg 8281 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga 8341 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata 8401 aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc 8461 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag 8521 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac 8581 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc 8641 atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga 8701 gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt 8761 acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc 8821 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg 8881 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag 8941 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact 9001 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg 9061 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc 9121 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg 9181 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg 9241 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag 9301 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc 9361 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct 9421 ttttacggtt cctggccttt tgctggcctt ttgctcacat gtt //