LOCUS Exported 9159 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION MCP-3XBFPnls. ACCESSION . VERSION . KEYWORDS pHAGE-TO-MCP-3XBFPnls SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9159) AUTHORS Ma H, Tu LC, Naseri A, Huisman M, Zhang S, Grunwald D, Pederson T TITLE Multiplexed labeling of genomic loci with dCas9 and engineered sgRNAs using CRISPRainbow. JOURNAL Nat Biotechnol. 2016 Apr 18. doi: 10.1038/nbt.3526. PUBMED 27088723 REFERENCE 2 (bases 1 to 9159) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9159 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 1..634 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 681..806 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1303..1536 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1721..1765 /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 2058..2175 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" enhancer 2299..2602 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2603..2806 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2757..2777 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 2808..2826 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2829..2847 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 2857..2881 /label=LNCX /note="Human CMV promoter, forward primer" regulatory 2962..2971 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3343..4053 /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" CDS 4072..4782 /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" CDS 4801..5511 /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" CDS 5518..5538 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" misc_feature 5549..6137 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5602..5622) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6020..6031) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 6212..6445 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" primer_bind complement(6564..6582) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 6650..6754 /gene="bla" /label=AmpR promoter CDS 6755..7615 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(6973..6992) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7786..8374 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8275..8294 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 8528..8545 /label=L4440 /note="L4440 vector, forward primer" primer_bind 8637..8656 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" protein_bind 8785..8806 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 8821..8851 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8859..8875 /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)." primer_bind 8864..8886 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 8883..8899 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 8883..8899 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 8949..8968 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttgacaaatg 2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa 2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca 2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagcgat 2221 gctagctgat gcgggcccta gttattaata gtaatcaatt acggggtcat tagttcatag 2281 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc 2341 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg 2401 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca 2461 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc 2521 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt 2581 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata 2641 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt 2701 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca 2761 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctcc ctatcagtga 2821 tagagatctc cctatcagtg atagagatcg tcgacgagct cgtttagtga accgtcagat 2881 cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg accgatccag 2941 cctaccggtg gaagtgggcg cgccaccatg gcttctaact ttactcagtt cgttctcgtc 3001 gacaatggcg gaactggcga cgtgactgtc gccccaagca acttcgctaa cgggatcgct 3061 gaatggatca gctctaactc gcgttcacag gcttacaaag taacctgtag cgttcgtcag 3121 agctctgcgc agaatcgcaa atacaccatc aaagtcgagg tgcctaaagg cgcctggcgt 3181 tcgtacttaa atatggaact aaccattcca attttcgcca cgaattccga ctgcgagctt 3241 attgttaagg caatgcaagg tctcctaaaa gatggaaacc cgattccctc agcaatcgca 3301 gcaaactccg gcatctacac tagtggctct ggatccggaa gtatggtgtc taagggcgaa 3361 gagctgatta aggagaacat gcacatgaag ctgtacatgg agggcaccgt ggacaaccat 3421 cacttcaagt gcacatccga gggcgaaggc aagccctacg agggcaccca gaccatgaga 3481 atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg acatcctggc tactagcttc 3541 ctctacggca gcaagacctt catcaaccac acccagggca tccccgactt cttcaagcag 3601 tccttccctg agggcttcac atgggagaga gtcaccacat acgaagacgg gggcgtgctg 3661 accgctaccc aggacaccag cctccaggac ggctgcctca tctacaacgt caagatcaga 3721 ggggtgaact tcacatccaa cggccctgtg atgcagaaga aaacactcgg ctgggaggcc 3781 ttcaccgaga cgctgtaccc cgctgacggc ggcctggaag gcagaaacga catggccctg 3841 aagctcgtgg gcgggagcca tctgatcgca aacgccaaga ccacatatag atccaagaaa 3901 cccgctaaga acctcaagat gcctggcgtc tactatgtgg actacagact ggaaagaatc 3961 aaggaggcca acaacgaaac ctacgtcgag cagcacgagg tggcagtggc cagatactgc 4021 gacctcccta gcaaactggg gcacaagctt aattctggcg gccgctctgg catggtgtct 4081 aagggcgaag agctgattaa ggagaacatg cacatgaagc tgtacatgga gggcaccgtg 4141 gacaaccatc acttcaagtg cacatccgag ggcgaaggca agccctacga gggcacccag 4201 accatgagaa tcaaggtggt cgagggcggc cctctcccct tcgccttcga catcctggct 4261 actagcttcc tctacggcag caagaccttc atcaaccaca cccagggcat ccccgacttc 4321 ttcaagcagt ccttccctga gggcttcaca tgggagagag tcaccacata cgaagacggg 4381 ggcgtgctga ccgctaccca ggacaccagc ctccaggacg gctgcctcat ctacaacgtc 4441 aagatcagag gggtgaactt cacatccaac ggccctgtga tgcagaagaa aacactcggc 4501 tgggaggcct tcaccgagac gctgtacccc gctgacggcg gcctggaagg cagaaacgac 4561 atggccctga agctcgtggg cgggagccat ctgatcgcaa acgccaagac cacatataga 4621 tccaagaaac ccgctaagaa cctcaagatg cctggcgtct actatgtgga ctacagactg 4681 gaaagaatca aggaggccaa caacgaaacc tacgtcgagc agcacgaggt ggcagtggcc 4741 agatactgcg acctccctag caaactgggg cacaagctta atggtagcat cgatggtagc 4801 atggtgtcta agggcgaaga gctgattaag gagaacatgc acatgaagct gtacatggag 4861 ggcaccgtgg acaaccatca cttcaagtgc acatccgagg gcgaaggcaa gccctacgag 4921 ggcacccaga ccatgagaat caaggtggtc gagggcggcc ctctcccctt cgccttcgac 4981 atcctggcta ctagcttcct ctacggcagc aagaccttca tcaaccacac ccagggcatc 5041 cccgacttct tcaagcagtc cttccctgag ggcttcacat gggagagagt caccacatac 5101 gaagacgggg gcgtgctgac cgctacccag gacaccagcc tccaggacgg ctgcctcatc 5161 tacaacgtca agatcagagg ggtgaacttc acatccaacg gccctgtgat gcagaagaaa 5221 acactcggct gggaggcctt caccgagacg ctgtaccccg ctgacggcgg cctggaaggc 5281 agaaacgaca tggccctgaa gctcgtgggc gggagccatc tgatcgcaaa cgccaagacc 5341 acatatagat ccaagaaacc cgctaagaac ctcaagatgc ctggcgtcta ctatgtggac 5401 tacagactgg aaagaatcaa ggaggccaac aacgaaacct acgtcgagca gcacgaggtg 5461 gcagtggcca gatactgcga cctccctagc aaactggggc acaagcttaa tctcgagcca 5521 aagaaaaagc ggaaagtgta atctagataa tcaacctctg gattacaaaa tttgtgaaag 5581 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat 5641 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc 5701 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg 5761 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct 5821 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct 5881 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg 5941 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac 6001 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct 6061 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct 6121 ttgggccgcc tccccgcctg agatccttta agaccaatga cttacaaggc agctgtagat 6181 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga 6241 caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag 6301 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt 6361 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt 6421 tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta 6481 taacttgcaa agaaatgaat atcagagagt gagaggcccg ggttaattaa ggaaagggct 6541 agatcattct tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat 6601 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc 6661 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg 6721 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc 6781 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt 6841 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct 6901 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac 6961 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact 7021 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa 7081 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga 7141 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt 7201 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga 7261 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg 7321 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat 7381 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat 7441 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc 7501 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga 7561 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc 7621 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag 7681 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc 7741 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt 7801 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt 7861 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat 7921 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc 7981 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa 8041 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg 8101 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag 8161 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag 8221 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa 8281 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt 8341 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg 8401 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc 8461 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac 8521 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct 8581 ccccgcgcgt tggccgattc attaatgcag caagctcatg gctgactaat tttttttatt 8641 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt 8701 tttggaggcc taggcttttg caaaaagctc cccgtggcac gacaggtttc ccgactggaa 8761 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc 8821 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca 8881 cacaggaaac agctatgaca tgattacgaa tttcacaaat aaagcatttt tttcactgca 8941 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaactggat 9001 aactcaagct aaccaaaatc atcccaaact tcccacccca taccctatta ccactgccaa 9061 ttacctgtgg tttcatttac tctaaacctg tgattcctct gaattatttt cattttaaag 9121 aaattgtatt tgttaaatat gtactacaaa cttagtagt //