LOCUS Exported 8605 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses the fluorescent protein TagBFP in a lentiviral backbone. ACCESSION . VERSION . KEYWORDS pLenti6.2_TagBFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8605) TITLE Fluorescent protein expressing plasmids suitable for lentiviral approaches JOURNAL Unpublished REFERENCE 2 (bases 1 to 8605) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8605 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 4..108 /gene="bla" /label=AmpR promoter CDS 109..969 /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 complement(327..346) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1140..1728 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1629..1648 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 1882..1899 /label=L4440 /note="L4440 vector, forward primer" protein_bind 2016..2037 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 2052..2082 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2090..2106 /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 2095..2117 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 2114..2130 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 2114..2130 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 2149..2169 /label=T3 /note="T3 promoter, forward primer" promoter 2151..2169 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 2197..2423 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 2424..2604 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 2651..2776 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 3269..3502 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 3687..3731 /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 3998..4114 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" enhancer 4137..4440 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4441..4644 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4595..4615 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 4641..4665 /label=LNCX /note="Human CMV promoter, forward primer" CDS 4744..5445 /codon_start=1 /product="monomeric blue fluorescent protein" /label=TagBFP /note="mammalian codon-optimized" /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC DLPSKLGHKLN" misc_feature 5461..6049 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5514..5534) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5932..5943) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" promoter 6063..6562 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(6098..6121) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 6495..6514 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 6583..7182 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(6583..6602) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 7079..7099 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 7320..7553 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 7625..7746 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(7662..7681) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 7716..7735 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 7786..7921 /label=SV40 ori /note="SV40 origin of replication" primer_bind 7848..7867 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(7941..7960) /label=T7 /note="T7 promoter, forward primer" promoter complement(7942..7960) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(7970..7987) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(7970..7986) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7979..8001) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 8128..8583 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(8215..8234) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 8425..8446 /label=F1ori-F /note="F1 origin, forward primer" ORIGIN 1 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg 61 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa 121 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac 181 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac 241 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt 301 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc 361 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca 421 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc 481 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag 541 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa 601 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg 661 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa 721 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg 781 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt 841 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt 901 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag 961 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat 1021 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct 1081 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct 1141 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca 1201 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc 1261 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc 1321 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct 1381 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag 1441 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc 1501 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg 1561 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag 1621 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt 1681 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac 1741 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg 1801 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc 1861 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata 1921 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt 1981 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag 2041 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga 2101 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct 2161 cactaaaggg aacaaaagct ggagctgcaa gcttaatgta gtcttatgca atactcttgt 2221 agtcttgcaa catggtaacg atgagttagc aacatgcctt acaaggagag aaaaagcacc 2281 gtgcatgccg attggtggaa gtaaggtggt acgatcgtgc cttattagga aggcaacaga 2341 cgggtctgac atggattgga cgaaccactg aattgccgca ttgcagagat attgtattta 2401 agtgcctagc tcgatacata aacgggtctc tctggttaga ccagatctga gcctgggagc 2461 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc 2521 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt 2581 agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa 2641 accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga 2701 ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga 2761 tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg 2821 gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga 2881 gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat 2941 actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa 3001 tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc 3061 tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc 3121 tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat 3181 ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg 3241 tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag 3301 caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt 3361 ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt 3421 tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat 3481 acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca 3541 ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca 3601 cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa 3661 ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat 3721 gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata aaattattca 3781 taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga 3841 atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg 3901 gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca 3961 ttcgattagt gaacggatct cgacggtatc ggttaacttt taaaagaaaa ggggggattg 4021 gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag 4081 aattacaaaa acaaattaca aaattcaaaa ttttatcgat aagcttggga gttccgcgtt 4141 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg 4201 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg 4261 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt 4321 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg 4381 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg 4441 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt 4501 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac 4561 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg 4621 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat 4681 ccacgctgtt ttgacctcca tagaagacac cgactctaga gatatcttcg aaggcgcgcc 4741 accatgagcg agctgattaa ggagaacatg cacatgaagc tgtacatgga gggcaccgtg 4801 gacaaccatc acttcaagtg cacatccgag ggcgaaggca agccctacga gggcacccag 4861 accatgagaa tcaaggtggt cgagggcggc cctctcccct tcgccttcga catcctggct 4921 actagcttcc tctacggcag caagaccttc atcaaccaca cccagggcat ccccgacttc 4981 ttcaagcagt ccttccctga gggcttcaca tgggagagag tcaccacata cgaagacggg 5041 ggcgtgctga ccgctaccca ggacaccagc ctccaggacg gctgcctcat ctacaacgtc 5101 aagatcagag gggtgaactt cacatccaac ggccctgtga tgcagaagaa aacactcggc 5161 tgggaggcct tcaccgagac gctgtacccc gctgacggcg gcctggaagg cagaaacgac 5221 atggccctga agctcgtggg cgggagccat ctgatcgcaa acatcaagac cacatataga 5281 tccaagaaac ccgctaagaa cctcaagatg cctggcgtct actatgtgga ctacagactg 5341 gaaagaatca aggaggccaa caacgagacc tacgtcgagc agcacgaggt ggcagtggcc 5401 agatactgcg acctccctag caaactgggg cacaagctta attaaagcgg ccgcgtcgac 5461 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct 5521 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt 5581 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg 5641 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact 5701 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct 5761 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg 5821 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc 5881 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc 5941 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt 6001 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaattcta 6061 ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct 6121 gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag 6181 gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt 6241 caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc 6301 acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct 6361 ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag 6421 gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc 6481 ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc 6541 ttcctcatct ccgggccttt cgacctgcag cccaagctta ccatgaccga gtacaagccc 6601 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg 6661 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc 6721 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc 6781 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg 6841 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag 6901 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc 6961 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc 7021 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc 7081 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa 7141 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacgcccgcc ccacgacccg 7201 cagcgcccga ccgaaaggag cgcacgaccc catgcatctc gagggcccgg tacctttaag 7261 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact 7321 ggaagggcta gctcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct 7381 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa 7441 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc 7501 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag 7561 ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga 7621 gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 7681 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 7741 atcttatcat gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact 7801 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 7861 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 7921 ctagggacgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg 7981 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac 8041 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac 8101 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg 8161 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt 8221 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc 8281 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg 8341 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga 8401 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc 8461 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa 8521 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa 8581 tttaggtggc acttttcggg gaaat //