LOCUS Exported 8865 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION piggyBac vector of 15xQUAS-mCD8:GFP; used for mosquito tranformation. ACCESSION . VERSION . KEYWORDS pBAC-ECFP-15xQUAS_TATA-mcd8-GFP-SV40 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8865) AUTHORS Riabinina O, Task D, Marr E, Lin CC, Alford R, O'Brochta DA, Potter CJ TITLE Organization of olfactory centres in the malaria mosquito Anopheles gambiae. JOURNAL Nat Commun. 2016 Oct 3;7:13010. doi: 10.1038/ncomms13010. PUBMED 27694947 REFERENCE 2 (bases 1 to 8865) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8865 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 1..17 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" repeat_region 147..209 /label=piggyBac right (3') inverted repeat /note="piggyBac transposon-specific inverted terminal repeat sequence (ITR)" regulatory 695..704 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 701..1420 /codon_start=1 /product="enhanced CFP" /label=ECFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(746..767) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1007..1026) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1354..1375 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 1543..1664 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" promoter 2076..2314 /label=hsp70 promoter /note="Drosophila melanogaster hsp70Bb promoter" primer_bind 2261..2282 /label=pCasper-hs /note="Drosophila Hsp70 promoter, forward primer" CDS 2390..3055 /codon_start=1 /product="mouse lymphocyte antigen CD8 alpha chain" /label=mCD8 /translation="MASPLTRFLSLNLLLLGESIILGSGEAKPQAPELRIFPKKMDAEL GQKVDLVCEVLGSVSQGCSWLFQNSSSKLPQPTFVVYMASSHNKITWDEKLNSSKLFSA MRDTNNKYVLTLNKFSKENEGYYFCSVISNSVMYFSSVVPVLQKVNSTTTKPVLRTPSP VHPTGTSQPQRPEDCRPRGSVKGTGLDFACDIYIWAPLAGICVALLLSLIITLICYHSR " CDS 3062..3775 /codon_start=1 /product="green fluorescent protein" /label=GFP /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" primer_bind complement(3092..3120) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 3715..3735 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" intron 3862..3927 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 4057..4077 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 4349..4483 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" intron 4650..4715 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 4845..4865 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 5137..5271 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind 5292..5308 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 5293..5309 /label=KS primer /note="common sequencing primer, one of multiple similar variants" repeat_region 5592..5626 /label=piggyBac left (5') inverted repeat /note="piggyBac transposon-specific inverted terminal repeat sequence (ITR)" primer_bind complement(6117..6133) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6117..6133) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6130..6152) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6141..6157 /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(6165..6195) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6210..6231 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6348..6365) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6519..7107) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6599..6618) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7278..8138) /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 7901..7920 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8139..8243) /gene="bla" /label=AmpR promoter rep_origin complement(8269..8724) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(8406..8427) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 8618..8637 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 8851..8 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 8865..17 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" ORIGIN 1 gtaaaacgac ggccagtgag cgcgcctcgt tcattcacgt ttttgaaccc gtggaggacg 61 ggcagactcg cggtgcaaat gtgttttaca gcgtgatgga gcagatgaag atgctcgaca 121 cgctgcagaa cacgcagcta gattaaccct agaaagataa tcatattgtg acgtacgtta 181 aagataatca tgtgtaaaat tgacgcatgt gttttatcgg tctgtatatc gaggtttatt 241 tattaatttg aatagatatt aagttttatt atatttacac ttacatacta ataataaatt 301 caacaaacaa tttatttatg tttatttatt tattaaaaaa aacaaaaact caaaatttct 361 tctataaagt aacaaaactt ttatcgaatt cctgcagccc gggggatcca ctagttctag 421 tgttcccaca atggttaatt cgagctcgcc cggggatcta attcaattag agactaattc 481 aattagagct aattcaatta ggatccaagc ttatcgattt cgaaccctcg accgccggag 541 tataaataga ggcgcttcgt ctacggagcg acaattcaat tcaaacaagc aaagtgaaca 601 cgtcgctaag cgaaagctaa gcaaataaac aagcgcagct gaacaagcta aacaatcggg 661 gtaccgctag agtcgacggt acgatccacc ggtcgccacc atggtgagca agggcgagga 721 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa 781 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt 841 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgacctg 901 gggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc 961 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta 1021 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa 1081 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacat 1141 cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa 1201 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac 1261 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc 1321 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc 1381 cgccgggatc actctcggca tggacgagct gtacaagtaa agcggccgcg actctagatc 1441 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc 1501 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct 1561 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca 1621 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaagctt atcgatacgc 1681 gtacggcact agagcggccg ccaccgcggt cacgtgtcac tgggtcagtg ggtaatcgct 1741 tatcctcgga taaacaatta tcctcacggg taatcgctta tccgctcggg taatcgctta 1801 tcctcgggta atcgcttatc cttgggtaat cgcttatcct cggataaaca attatcctca 1861 cgggtaatcg cttatccgct cgggtaatcg cttatcctcg ggtaatcgct tatccttggg 1921 taatcgctta tcctcggata aacaattatc ctcacgggta atcgcttatc cgctcgggta 1981 atcgcttatc ctcgggtaat cgcttatcct ttcacgttgg gactcagtga ggaggacctg 2041 aattcctgca gcccgagcgg agactctagc gagcgccgga gtataaatag aggcgcttcg 2101 tctacggagc gacaattcaa ttcaaacaag caaagtgaac acgtcgctaa gcgaaagcta 2161 agcaaataaa caagcgcagc tgaacaagct aaacaatctg cagtaaagtg caagttaaag 2221 tgaatcaatt aaaagtaacc agcaaccaag taaatcaact gcaactactg aaatctgcca 2281 agaagtaatt attgaataca agaagagaac tctgaatagg gaattgggaa ttcgttaaca 2341 gatcggggga tcaattcgtt aacagatctg cggccgcggc tcgagcaaaa tggcctcacc 2401 gttgacccgc tttctgtcgc tgaacctgct gctgctgggt gagtcgatta tcctggggag 2461 tggagaagct aagccacagg cacccgaact ccgaatcttt ccaaagaaaa tggacgccga 2521 acttggtcag aaggtggacc tggtatgtga agtgttgggg tccgtttcgc aaggatgctc 2581 ttggctcttc cagaactcca gctccaaact cccccagccc accttcgttg tctatatggc 2641 ttcatcccac aacaagataa cgtgggacga gaagctgaat tcgtcgaaac tgttttctgc 2701 catgagggac acgaataata agtacgttct caccctgaac aagttcagca aggaaaacga 2761 aggctactat ttctgctcag tcatcagcaa ctcggtgatg tacttcagtt ctgtcgtgcc 2821 agtccttcag aaagtgaact ctactactac caagccagtg ctgcgaactc cctcacctgt 2881 gcaccctacc gggacatctc agccccagag accagaagat tgtcggcccc gtggctcagt 2941 gaaggggacc ggattggact tcgcctgtga tatttacatc tgggcaccct tggccggaat 3001 ctgcgtggcc cttctgctgt ccttgatcat cactctcatc tgctaccaca gccgcggatc 3061 catgagtaaa ggagaagaac ttttcactgg agttgtccca attcttgttg aattagatgg 3121 tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg caacatacgg 3181 aaaacttacc cttaaattta tttgcactac tggaaaacta cctgttccat ggccaacact 3241 tgtcactact ttaacttatg gtgttcaatg cttttcaaga tacccagatc atatgaaaca 3301 gcatgacttt ttcaagagtg ccatgcccga aggttatgtc caggaaagaa ctatattttt 3361 caaagatgac gggaactaca agacacgtgc tgaagtcaag tttgaaggtg atacccttgt 3421 taatagaatc gagttaaaag gtattgattt taaagaagat ggaaacattc ttggacacaa 3481 attggaatac aactataact cacacaatgt atacatcatg gcagacaaac aaaagaatgg 3541 aatcaaagtt aacttcaaaa ttagacacaa cattgaagat ggaagcgttc aactagcaga 3601 ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta 3661 cctgtccaca caatctgccc tttcgaaaga tcccaacgaa aagagagacc acatggtcct 3721 tcttgagttt gtaacagctg ctgggattac acatggcatg gatgaactat acaaataatc 3781 tagaggatct ttgtgaagga accttacttc tgtggtgtga cataattgga caaactacct 3841 acagagattt aaagctctaa ggtaaatata aaatttttaa gtgtataatg tgttaaacta 3901 ctgattctaa ttgtttgtgt attttagatt ccaacctatg gaactgatga atgggagcag 3961 tggtggaatg cctttaatga ggaaaacctg ttttgctcag aagaaatgcc atctagtgat 4021 gatgaggcta ctgctgactc tcaacattct actcctccaa aaaagaagag aaaggtagaa 4081 gaccccaagg actttccttc agaattgcta agttttttga gtcatgctgt gtttagtaat 4141 agaactcttg cttgctttgc tatttacacc acaaaggaaa aagctgcact gctatacaag 4201 aaaattatgg aaaaatattt gatgtatagt gccttgacta gagatcataa tcagccatac 4261 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa 4321 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa 4381 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 4441 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccactaga aggccttagt 4501 atggggtacc ccaagggcga attctgcaga tatccatcac actggcggcc gcggctcgag 4561 ggtacctcta gaggatcttt gtgaaggaac cttacttctg tggtgtgaca taattggaca 4621 aactacctac agagatttaa agctctaagg taaatataaa atttttaagt gtataatgtg 4681 ttaaactact gattctaatt gtttgtgtat tttagattcc aacctatgga actgatgaat 4741 gggagcagtg gtggaatgcc tttaatgagg aaaacctgtt ttgctcagaa gaaatgccat 4801 ctagtgatga tgaggctact gctgactctc aacattctac tcctccaaaa aagaagagaa 4861 aggtagaaga ccccaaggac tttccttcag aattgctaag ttttttgagt catgctgtgt 4921 ttagtaatag aactcttgct tgctttgcta tttacaccac aaaggaaaaa gctgcactgc 4981 tatacaagaa aattatggaa aaatatttga tgtatagtgc cttgactaga gatcataatc 5041 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg 5101 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat 5161 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat 5221 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat ccactagaag 5281 gcctagatct ctcgaggtcg acggtatcga taagcttgat atctataaca agaaaatata 5341 tatataataa gttatcacgt aagtagaaca tgaaataaca atataattat cgtatgagtt 5401 aaatcttaaa agtcacgtaa aagataatca tgcgtcattt tgactcacgc ggtcgttata 5461 gttcaaaatc agtgacactt accgcattga caagcacgcc tcacgggagc tccaagcggc 5521 gactgagatg tcctaaatgc acagcgacgg attcgcgcta tttagaaaga gagagcaata 5581 tttcaagaat gcatgcgtca attttacgca gactatcttt ctagggttaa tctagctgca 5641 tcaggatcat atcgtcgggt cttttttccg gctcagtcat cgcccaagct ggcgctatct 5701 gggcatcggg gaggaagaag cccgtgcctt ttcccgcgag gttgaagcgg catggaaaga 5761 gtttgccgag gatgactgct gctgcattga cgttgagcga aaacgcacgt ttaccatgat 5821 gattcgggaa ggtgtggcca tgcacgcctt taacggtgaa ctgttcgttc aggccacctg 5881 ggataccagt tcgtcgcggc ttttccggac acagttccgg atggtcagcc cgaagcgcat 5941 cagcaacccg aacaataccg gcgacagccg gaactgccgt gccggtgtgc agattaatga 6001 cagcggtgcg gcgctgggat attacgtcag cgaggacggg tatcctggct ggatgccgca 6061 gaaatggaca tggatacccc gtgagttacc cggcgggcgc gcttggcgta atcatggtca 6121 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga 6181 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg 6241 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc 6301 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac 6361 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata 6421 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa 6481 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 6541 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 6601 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 6661 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca 6721 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 6781 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 6841 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 6901 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga 6961 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 7021 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 7081 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 7141 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc 7201 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag 7261 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt 7321 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag 7381 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca 7441 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact 7501 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca 7561 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg 7621 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc 7681 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg 7741 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca 7801 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt 7861 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc 7921 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc 7981 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca 8041 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa 8101 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat 8161 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa 8221 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg 8281 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat 8341 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg 8401 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc 8461 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt 8521 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag 8581 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg 8641 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc 8701 ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag 8761 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa 8821 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgtt //