LOCUS Exported 8238 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA3-TRIF-CFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8238) TITLE TLR Signaling Plasmids REFERENCE 2 (bases 1 to 8238) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8238 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 1101..1112 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" CDS 3055..3774 /codon_start=1 /product="cyan variant of GFP" /label=CFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3100..3121) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3361..3380) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3708..3729 /label=EGFP-C /note="EGFP, forward primer" promoter complement(3790..3808) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(3791..3808) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(3828..3845) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3834..4058 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 4104..4532 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4191..4210) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4401..4422 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4541..4561) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4546..4876 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4727..4862 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4789..4808 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4943..5737 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4997..5016) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5607..5626 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5911..6032 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5948..5967) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6002..6021 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(6081..6097) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6081..6097) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6094..6116) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6105..6121 /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(6129..6159) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6174..6195 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6312..6329) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6483..7071) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6563..6582) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7242..8102) /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 7865..7884 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8103..8207) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc 901 gagctcggat ccatggcctg cacaggccca tcacttccta gcgccttcga cattctaggt 961 gcagcaggcc aggacaagct cttgtatctg aagcacaaac tgaagacccc acgcccaggc 1021 tgccaggggc aggacctcct gcatgccatg gttctcctga agctgggcca ggaaactgag 1081 gccaggatct ctctagaggc attgaaggcc gatgcggtgg cccggctggt ggcccgccag 1141 tgggctggcg tggacagcac cgaggaccca gaggagcccc cagatgtgtc ctgggctgtg 1201 gcccgcttgt accacctgct ggctgaggag aagctgtgcc ccgcctcgct gcgggacgtg 1261 gcctaccagg aagccgtccg caccctcagc tccagggacg accaccggct gggggaactt 1321 caggatgagg cccgaaaccg gtgtgggtgg gacattgctg gggatccagg gagcatccgg 1381 acgctccagt ccaatctggg ctgcctccca ccatcctcgg ctttgccctc tgggaccagg 1441 agcctcccac gccccattga cggtgtttcg gactggagcc aagggtgctc cctgcgatcc 1501 actggcagcc ctgcctccct ggccagcaac ttggaaatca gccagtcccc taccatgccc 1561 ttcctcagcc tgcaccgcag cccacatggg cccagcaagc tctgtgacga cccccaggcc 1621 agcttggtgc ccgagcctgt ccccggtggc tgccaggagc ctgaggagat gagctggccg 1681 ccatcggggg agattgccag cccaccagag ctgccaagca gcccacctcc tgggcttccc 1741 gaagtggccc cagatgcaac ctccactggc ctccctgata cccccgcagc tccagaaacc 1801 agcaccaact acccagtgga gtgcaccgag gggtctgcag gcccccagtc tctccccttg 1861 cctattctgg agccggtcaa aaacccctgc tctgtcaaag accagacgcc actccaactt 1921 tctgtagaag ataccacctc tccaaatacc aagccgtgcc cacctactcc caccacccca 1981 gaaacatccc ctcctcctcc tcctcctcct ccttcatcta ctccttgttc agctcacctg 2041 accccctcct ccctgttccc ttcctccctg gaatcatcat cggaacagaa attctataac 2101 tttgtgatcc tccacgccag ggcagacgaa cacatcgccc tgcgggttcg ggagaagctg 2161 gaggcccttg gcgtgcccga cggggccacc ttctgtgagg atttccaggt gccggggcgc 2221 ggggagctga gctgcctgca ggacgccata gaccactcag ctttcatcat cctacttctc 2281 acctccaact tcgactgtcg cctgagcctg caccaggtga accaagccat gatgagcaac 2341 ctcacgcgac aggggtcgcc agactgtgtc atccccttcc tgcccctgga gagctccccg 2401 gcccagctca gctccgacac ggccagcctg ctctccgggc tggtgcggct ggacgaacac 2461 tcccagatct tcgccaggaa ggtggccaac accttcaagc cccacaggct tcaggcccga 2521 aaggccatgt ggaggaagga acaggacacc cgagccctgc gggaacagag ccaacacctg 2581 gacggtgagc ggatgcaggc ggcggcactg aacgcagcct actcagccta cctccagagc 2641 tacttgtcct accaggcaca gatggagcag ctccaggtgg cttttgggag ccacatgtca 2701 tttgggactg gggcgcccta tggggctcga atgccctttg ggggccaggt gcccctggga 2761 gccccgccac cctttcccac ttggccgggg tgcccgcagc cgccacccct gcacgcatgg 2821 caggctggca cccccccacc gccctcccca cagccagcag cctttccaca gtcactgccc 2881 ttcccgcagt ccccagcctt ccctacggcc tcacccgcac cccctcagag cccagggctg 2941 caacccctca ttatccacca cgcacagatg gtacagctgg ggctgaacaa ccacatgtgg 3001 aaccagagag ggtcccaggc gcccgaggac aagacgcagg aggcagaact cgagatggtg 3061 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac 3121 gtaaacggcc acaggttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag 3181 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg 3241 accaccctga cctggggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac 3301 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgtaccat cttcttcaag 3361 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac 3421 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg 3481 gagtacaact acatcagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc 3541 aaggcccact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac 3601 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg 3661 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg 3721 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaatctaga 3781 gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct cgactgtgcc 3841 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg 3901 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag 3961 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga 4021 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag 4081 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt 4141 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc 4201 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg 4261 catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta 4321 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt 4381 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat 4441 ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa 4501 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg 4561 tgtggaaagt ccccaggctc cccaggcagg cagaagtatg caaagcatgc atctcaatta 4621 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat 4681 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac 4741 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga 4801 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg 4861 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga 4921 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc 4981 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga 5041 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct 5101 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac 5161 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct 5221 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt 5281 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt 5341 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt 5401 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag 5461 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt 5521 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg 5581 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg 5641 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg 5701 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg 5761 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat 5821 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg 5881 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac 5941 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt 6001 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc 6061 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca 6121 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg 6181 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg 6241 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc 6301 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 6361 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 6421 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 6481 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 6541 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 6601 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 6661 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 6721 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 6781 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 6841 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 6901 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt 6961 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 7021 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 7081 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 7141 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 7201 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 7261 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 7321 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 7381 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 7441 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 7501 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 7561 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 7621 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 7681 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 7741 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 7801 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 7861 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 7921 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 7981 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 8041 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 8101 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 8161 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 8221 aaagtgccac ctgacgtc //