LOCUS Exported 8195 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Constitutive expression of mTagBFP2-Dectin1A-N-10 using PGK promoter in a lentiviral backbone.. ACCESSION . VERSION . KEYWORDS pRRlsinPGK_dectintagBFP2_WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8195) TITLE Torok-Storb lab lentiviral fluorescent protein JOURNAL Unpublished REFERENCE 2 (bases 1 to 8195) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8195 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(29..46) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(200..788) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(280..299) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(959..1819) /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 1582..1601 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1820..1924) /gene="bla" /label=AmpR promoter rep_origin complement(1950..2405) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2087..2108) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2299..2318 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2532..2554 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2546..2563 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2547..2563 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 2573..2592 /label=T7 /note="T7 promoter, forward primer" promoter 2573..2591 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" rep_origin 2612..2747 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(2666..2685) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 2787..2908 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2798..2817) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2852..2871 /label=SV40pA-R /note="SV40 polyA, reverse primer" LTR 2980..3213 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" misc_feature 3300..3888 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" CDS 3406..3417 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind 3815..3835 /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3950..4660) /codon_start=1 /product="enhanced monomeric blue fluorescent protein (Subach et al., 2011)" /label=mTagBFP2 /note="mammalian codon-optimized" /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV ARYCDLPSKLGHKLN" regulatory 4657..4666 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 5428..5437 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" promoter complement(5452..5962) /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" primer_bind complement(5515..5533) /label=hPGK-F /note="Human PGK promoter, forward primer" misc_feature 6008..6125 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" CDS complement(6392..6436) /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 6621..6854 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." misc_feature 7347..7472 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" LTR 7519..7699 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" promoter complement(7700..7926) /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" primer_bind complement(7954..7974) /label=T3 /note="T3 promoter, forward primer" promoter complement(7954..7972) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(7993..8009) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7993..8009) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(8006..8028) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 8017..8033 /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(8041..8071) /label=lac promoter /note="promoter for the E. coli lac operon" ORIGIN 1 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc 61 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag 121 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa 181 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc 241 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc 301 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg 361 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt 421 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc 481 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc 541 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag 601 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg 661 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa 721 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 781 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 841 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 901 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 961 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 1021 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 1081 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 1141 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 1201 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 1261 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 1321 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 1381 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 1441 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 1501 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 1561 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 1621 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 1681 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 1741 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 1801 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 1861 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 1921 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt 1981 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat 2041 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa 2101 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg 2161 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa 2221 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc 2281 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag 2341 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg 2401 cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc 2461 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac 2521 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga 2581 ctcactatag ggcgaattgg gtacgtccct aggcctccaa aaaagcctcc tcactacttc 2641 tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag 2701 ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg 2761 ggcgggatag ctagagccag acatgataag atacattgat gagtttggac aaaccacaac 2821 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt 2881 aaccattata agctgcaata aacaagttcc tctcactctc tgatattcat ttctttgcaa 2941 gttataaata ctgaataata agatgacatg aactactact gctagagatt ttccacactg 3001 actaaaaggg tctgagggat ctctagttac cagagtcaca caacagacgg gcacacacta 3061 cttgaagcac tcaaggcaag ctttattgag gcttaagcag tgggttccct agttagccag 3121 agagctccca ggctcagatc tggtctaacc agagagaccc agtacaagca aaaagcagat 3181 cttgtcttcg ttgggagtga attagccctt ccagtccccc cttttctttt aaaaagtggc 3241 taagatctac agctgccttg taagtcattg gtcttaaagg taccgagctc gaattccagg 3301 cggggaggcg gcccaaaggg agatccgact cgtctgaggg cgaaggcgaa gacgcggaag 3361 aggccgcaga gccggcagca ggccgcggga aggaaggtcc gctggattga gggccgaagg 3421 gacgtagcag aaggacgtcc cgcgcagaat ccaggtggca acacaggcga gcagccatgg 3481 aaaggacgtc agcttccccg acaacaccac ggaattgtca gtgcccaaca gccgagcccc 3541 tgtccagcag cgggcaaggc aggcggcgat gagttccgcc gtggcaatag ggagggggaa 3601 agcgaaagtc ccggaaagga gctgacaggt ggtggcaatg ccccaaccag tgggggttgc 3661 gtcagcaaac acagtgcaca ccacgccacg ttgcctgaca acgggccaca actcctcata 3721 aagagacagc aaccaggatt tatacaagga ggagaaaatg aaagccatac gggaagcaat 3781 agcatgatac aaaggcatta aagcagcgta tccacatagc gtaaaaggag caacatagtt 3841 aagaatacca gtcaatcttt cacaaatttt gtaatccaga ggttgattgt cgactgcagt 3901 tcgaacccgg gcatatgtct agaaccggtg gatcagtcgc ggccgcttaa ttaagcttgt 3961 gccccagttt gctagggagg tcgcagtatc tggccactgc cacctcgtgc tgctcgacgt 4021 aggtctcgtt gttggcctcc ttgattcttt ccagtctgta gtccacatag tagacgccag 4081 gcatcttgag gttcttagcg ggtttcttgg atctatatgt ggtcttggcg tttgcgatca 4141 gatggctccc gcccacgagc ttcagggcca tgtcgtttct gccttccagg ccgccgtcag 4201 cggggtacag cgtctcggtg aaggcctccc agccgagtgt tttcttctgc atcacagggc 4261 cgttggatgt gaagttcacc cctctgatct tgacgttgta gatgaggcag ccgtcctgga 4321 ggctggtgtc ctgggtagcg gtcagcacgc ccccgtcttc gtatgtggtg actctctccc 4381 atgtgaagcc ctcagggaag gactgcttga agaagtcggg gatgccctgg gtgtggttga 4441 tgaaggtctt gctgccgtag aggaagctag tagccaggat gtcgaaggcg aaggggagag 4501 ggccgccctc gaccaccttg attctcatgg tctgggtgcc ctcgtagggc ttgccttcgc 4561 cctcggatgt gcacttgaag tgatggttgt ccacggtgcc ctccatgtac agcttcatgt 4621 gcatgttctc cttaatcagc tcttcgccct tagacaccat ggtggcgacc ggtggatccc 4681 ctccggagct cattgaaaac ttcttctcac aaatactata tgagggcaca ctacacagtt 4741 ggtcataaat gactgacacg tgaatccata cacaatttgg agatgggttt tcttgggtag 4801 ctgtggttct gatctgaaat aagttagaag agaatgttga tccatcctcc cagagccatg 4861 gtacttcagt ctggggccga gaaaggccta tccaaaatga attatcaggt tgggaagaca 4921 cttgttttac tataaatccc aattcatttg agctgtctat ctttaggaga ttagagccca 4981 gttgccagca ttgtctttta cttccatccc aggaatttag tgacatgctg aatagataac 5041 agctcttctc atatataatc caattaggag gacaagggct ggaaagaacc cctgtggttt 5101 tgacagcttt ggtaggagtc acactgtctt ctaaagatga ttgtgtgggt tgactgtggt 5161 tctctttatt tcttgataga aagtagccat tctccaatgt gttgcttcct gaattggatc 5221 tccaaatagc catggtaccc aggaccacag ctatcaccag tattaccaag cataggattc 5281 ccaaaattac agcaatgagg cgccaaggag gagatgcagc acacgatcct ttctctgaaa 5341 caacagctat cctggtattg ctttgagagt cgaagtgtaa ttgagtatat ccatcttcat 5401 ccaaattttc taaatcagga tgatattcca tggtggcgct agcagatccc cctggggaga 5461 gaggtcggtg attcggtcaa cgagggagcc gactgccgac gtgcgctccg gaggcttgca 5521 gaatgcggaa caccgcgcgg gcaggaacag ggcccacact accgccccac accccgcctc 5581 ccgcaccgcc ccttcccggc cgctgctctc ggcgcgccct gctgagcagc cgctattggc 5641 cacagcccat cgcggtcggc gcgctgccat tgctccctgg cgctgtccgt ctgcgagggt 5701 actagtgaga cgtgcggctt ccgtttgtca cgtccggcac gccgcgaacc gcaaggaacc 5761 ttcccgactt aggggcggag caggaagcgt cgccgggggg cccacaaggg tagcggcgaa 5821 gatccgggtg acgctgcgaa cggacgtgaa gaatgtgcga gacccagggt cggcgccgct 5881 gcgtttcccg gaaccacgcc cagagcagcc gcgtccctgc gcaaacccag ggctgccttg 5941 gaaaaggcgc aaccccaacc ccgtggaatt cgatatcaag cttctcgagg ctagtctcgt 6001 gatcgataaa attttgaatt tttgtaattt gtttttgtaa ttctttagtt tgtatgtctg 6061 ttgctattat gtctactatt ctttcccctg cactgtaccc cccaatcccc ccttttcttt 6121 taaaagttaa ccgataccgt cgagatccgt tcactaatcg aatggatctg tctctgtctc 6181 tctctccacc ttcttcttct attccttcgg gcctgtcggg tcccctcggg gttgggaggt 6241 gggtctgaaa cgataatggt gaatatccct gcctaactct attcactata gaaagtacag 6301 caaaaactat tcttaaacct accaagcctc ctactatcat tatgaataat tttatatacc 6361 acagccaatt tgttatgtta aaccaattcc acaaacttgc ccatttatct aattccaata 6421 attcttgttc attcttttct tgctggtttt gcgattcttc aattaaggag tgtattaagc 6481 ttgtgtaatt gttaatttct ctgtcccact ccatccaggt cgtgtgattc caaatctgtt 6541 ccagagattt attactccaa ctagcattcc aaggcacagc agtggtgcaa atgagttttc 6601 cagagcaacc ccaaatcccc aggagctgtt gatcctttag gtatctttcc acagccagga 6661 ttcttgcctg gagctgcttg atgccccaga ctgtgagttg caacagatgc tgttgcgcct 6721 caatagccct cagcaaattg ttctgctgct gcactatacc agacaataat tgtctggcct 6781 gtaccgtcag cgtcattgac gctgcgccca tagtgcttcc tgctgctccc aagaacccaa 6841 ggaacaaagc tcctattccc actgctcttt tttctctctg caccactctt ctctttgcct 6901 tggtgggtgc tactcctaat ggttcaattt ttactacttt atatttatat aattcacttc 6961 tccaattgtc cctcatatct cctcctccag gtctgaagat cagcggccgc ttgctgtgcg 7021 gtggtcttac ttttgttttg ctcttcctct atcttgtcta aagcttcctt ggtgtctttt 7081 atctctatcc tttgatgcac acaatagagg gttgctactg tattatataa tgatctaagt 7141 tcttctgatc ctgtctgaag ggatggttgt agctgtccca gtatttgtct acagccttct 7201 gatgtttcta acaggccagg attaactgcg aatcgttcta gctccctgct tgcccatact 7261 atatgtttta atttatattt tttctttccc cctggcctta accgaatttt ttcccatcgc 7321 gatctaattc tcccccgctt aatactgacg ctctcgcacc catctctctc cttctagcct 7381 ccgctagtca aaatttttgg cgtactcacc agtcgccgcc cctcgcctct tgccgtgcgc 7441 gcttcagcaa gccgagtcct gcgtcgagag agctcctctg gtttcccttt cgctttcaag 7501 tccctgttcg ggcgccactg ctagagattt tccacactga ctaaaagggt ctgagggatc 7561 tctagttacc agagtcacac aacagacggg cacacactac ttgaagcact caaggcaagc 7621 tttattgagg cttaagcagt gggttcccta gttagccaga gagctcccag gctcagatct 7681 ggtctaacca gagagacccg tttatgtatc gagctaggca cttaaataca atatctctgc 7741 aatgcggcaa ttcagtggtt cgtccaatcc atgtcagacc cgtctgttgc cttcctaata 7801 aggcacgatc gtaccacctt acttccacca atcggcatgc acggtgcttt ttctctcctt 7861 gtaaggcatg ttgctaactc atcgttacca tgttgcaaga ctacaagagt attgcataag 7921 actacattaa gcttgcagct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt 7981 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 8041 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 8101 caccttaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 8161 gcattaatga atcggccaac gcgcggggag aggcg //