LOCUS Exported 8368 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION 3rd generation Lentiviral vector with bi-cistronic expression of the blue fluorescent protein AmCyan and the gene of interest.. ACCESSION . VERSION . KEYWORDS pUltra-Smurf SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8368) TITLE pUltra series JOURNAL Unpublished REFERENCE 2 (bases 1 to 8368) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8368 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 8..234 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 235..415 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 462..587 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1080..1313 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1498..1542 /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 1840..1957 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" protein_bind complement(2004..2037) /label=lox5171 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GTACACAT). lox5171 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." promoter 2069..3280 /label=UbC promoter /note="human ubiquitin C promoter" primer_bind 3117..3136 /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" CDS 3312..3998 /codon_start=1 /product="Anemonia majano cyan fluorescent protein" /label=AmCyan1 /note="mammalian codon-optimized" /translation="MALSNKFIGDDMKMTYHMDGCVNGHYFTVKGEGSGKPYEGTQTST FKVTMANGGPLAFSFDILSTVFMYGNRCFTAYPTSMPDYFKQAFPDGMSYERTFTYEDG GVATASWEISLKGNCFEHKSTFHGVNFPADGPVMAKKTTGWDPSFEKMTVCDGILKGDV TAFLMLQGGGNYRCQFHTSYKTKKPVTMPPNHVVEHRIARTDLDKGGNSVQLTEHAVAH ITSVVPF" CDS 4017..4073 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 4095..4148 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" protein_bind complement(4170..4203) /label=lox5171 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GTACACAT). lox5171 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." misc_feature 4215..4803 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4268..4288) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4686..4697) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 4894..5127 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 5199..5320 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5236..5255) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5290..5309 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 5360..5495 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5422..5441 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(5515..5534) /label=T7 /note="T7 promoter, forward primer" promoter complement(5516..5534) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5544..5561) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(5544..5560) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5553..5575) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 5702..6157 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5789..5808) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5999..6020 /label=F1ori-F /note="F1 origin, forward primer" promoter 6183..6287 /gene="bla" /label=AmpR promoter CDS 6288..7148 /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(6506..6525) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7319..7907 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7808..7827 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 8061..8078 /label=L4440 /note="L4440 vector, forward primer" protein_bind 8195..8216 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 8231..8261 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8269..8285 /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 8274..8296 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 8293..8309 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 8293..8309 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 8328..8348 /label=T3 /note="T3 promoter, forward primer" promoter 8330..8348 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" ORIGIN 1 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag 61 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg 121 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact 181 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct 241 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 301 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 361 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc 421 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc 481 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa 541 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg 601 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata 661 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc 721 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga 781 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc 841 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca 901 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg 961 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga 1021 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata 1081 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg 1141 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg 1201 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag 1261 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt 1321 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt 1381 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt 1441 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag 1501 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata 1561 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta 1621 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta 1681 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa 1741 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt 1801 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat 1861 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa 1921 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag 1981 agatccagtt tggttaatta acgataactt cgtataatgt gtactatacg aagttatctc 2041 gagattaacc cgtgtcggct ccagatctgg cctccgcgcc gggttttggc gcctcccgcg 2101 ggcgcccccc tcctcacggc gagcgctgcc acgtcagacg aagggcgcag cgagcgtcct 2161 gatccttccg cccggacgct caggacagcg gcccgctgct cataagactc ggccttagaa 2221 ccccagtatc agcagaagga cattttagga cgggacttgg gtgactctag ggcactggtt 2281 ttctttccag agagcggaac aggcgaggaa aagtagtccc ttctcggcga ttctgcggag 2341 ggatctccgt ggggcggtga acgccgatga ttatataagg acgcgccggg tgtggcacag 2401 ctagttccgt cgcagccggg atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt 2461 cacttggtga gtagcgggct gctgggctgg ccggggcttt cgtggccgcc gggccgctcg 2521 gtgggacgga agcgtgtgga gagaccgcca agggctgtag tctgggtccg cgagcaaggt 2581 tgccctgaac tgggggttgg ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa 2641 tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg 2701 cggcaagaac ccaaggtctt gaggccttcg ctaatgcggg aaagctctta ttcgggtgag 2761 atgggctggg gcaccatctg gggaccctga cgtgaagttt gtcactgact ggagaactcg 2821 gtttgtcgtc tgttgcgggg gcggcagtta tggcggtgcc gttgggcagt gcacccgtac 2881 ctttgggagc gcgcgccctc gtcgtgtcgt gacgtcaccc gttctgttgg cttataatgc 2941 agggtggggc cacctgccgg taggtgtgcg gtaggctttt ctccgtcgca ggacgcaggg 3001 ttcgggccta gggtaggctc tcctgaatcg acaggcgccg gacctctggt gaggggaggg 3061 ataagtgagg cgtcagtttc tttggtcggt tttatgtacc tatcttctta agtagctgaa 3121 gctccggttt tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc 3181 accttttgaa atgtaatcat ttgggtcaat atgtaatttt cagtgttaga ctagtaaatt 3241 gtccgctaaa ttctggccgt ttttggcttt tttgttagac gaagcttggg ctgcaggtcc 3301 gatccaccgg tatggccctg tccaacaagt tcatcggcga cgacatgaag atgacctacc 3361 acatggacgg ctgcgtgaac ggccactact tcaccgtgaa gggcgagggc agcggcaagc 3421 cctacgaggg cacccagacc tccaccttca aggtgaccat ggccaacggc ggccccctgg 3481 ccttctcctt cgacatcctg tccaccgtgt tcatgtacgg caaccgctgc ttcaccgcct 3541 accccaccag catgcccgac tacttcaagc aggccttccc cgacggcatg tcctacgaga 3601 gaaccttcac ctacgaggac ggcggcgtgg ccaccgccag ctgggagatc agcctgaagg 3661 gcaactgctt cgagcacaag tccaccttcc acggcgtgaa cttccccgcc gacggccccg 3721 tgatggccaa gaagaccacc ggctgggacc cctccttcga gaagatgacc gtgtgcgacg 3781 gcatcttgaa gggcgacgtg accgccttcc tgatgctgca gggcggcggc aactacagat 3841 gccagttcca cacctcctac aagaccaaga agcccgtgac catgcccccc aaccacgtgg 3901 tggagcaccg catcgccaga accgacctgg acaagggcgg caacagcgtg cagctgaccg 3961 agcacgccgt ggcccacatc acctccgtgg tgcccttcgt gtacaaggga tctggagcaa 4021 caaacttctc actactcaaa caagcaggtg acgtggagga gaatcccggg ccttctagag 4081 gatccggctc cggcgagggc aggggaagtc tactaacatg cggggacgtg gaggaaaatc 4141 ccggcccagc tagcgtgtga tcagaattca taacttcgta taatgtgtac tatacgaagt 4201 tatgtcgact cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc 4261 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg 4321 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc 4381 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg 4441 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg 4501 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga 4561 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct 4621 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg 4681 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc 4741 ctcttccgca tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc 4801 cgcctggaat taattcgagc tcggtacctt taagaccaat gacttacaag gcagctgtag 4861 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctacgtaac tcccaacgaa 4921 gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg 4981 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc 5041 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct 5101 tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt 5161 tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata 5221 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc 5281 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat 5341 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc 5401 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct 5461 attccagaag tagtgaggag gcttttttgg aggcctaggg acgtacccaa ttcgccctat 5521 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac 5581 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat 5641 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg 5701 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc 5761 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc 5821 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt 5881 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg 5941 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt 6001 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta 6061 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt 6121 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg 6181 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga 6241 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac 6301 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc 6361 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca 6421 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc 6481 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg 6541 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac 6601 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca 6661 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg 6721 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac 6781 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg 6841 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat 6901 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg 6961 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg 7021 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc 7081 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc 7141 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt 7201 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt 7261 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt 7321 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag 7381 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca 7441 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca 7501 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg 7561 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg 7621 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct 7681 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga 7741 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc 7801 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctcacttg 7861 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg 7921 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt 7981 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc 8041 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac 8101 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc 8161 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg 8221 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat 8281 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc 8341 actaaaggga acaaaagctg gagctgca //