LOCUS Exported 8316 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION 3rd generation Lentiviral vector with bi-cistronic expression of dtomato and the gene of interest.. ACCESSION . VERSION . KEYWORDS pUltra-Chili SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8316) TITLE pUltra series JOURNAL Unpublished REFERENCE 2 (bases 1 to 8316) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8316 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 236..413 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 457..582 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1075..1308 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1493..1537 /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 1835..1952 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2017..3228 /label=UbC promoter /note="human ubiquitin C promoter" primer_bind 3065..3084 /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" CDS 3271..3972 /codon_start=1 /product="dimeric variant of DsRed fluorescent protein (Shaner et al., 2004)" /label=dTomato /note="mammalian codon-optimized" /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH HLFLYGMDELYK" primer_bind complement(3404..3422) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 3672..3691 /label=mCherry-F /note="mCherry, forward primer" primer_bind 3866..3889 /label=DsRed1-C /note="DsRed1, forward primer" CDS 3982..4038 /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 4060..4113 /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" misc_feature 4157..4745 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4210..4230) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4628..4639) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 4836..5069 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 5141..5262 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5178..5197) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5232..5251 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 5302..5437 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5364..5383 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(5457..5476) /label=T7 /note="T7 promoter, forward primer" promoter complement(5458..5476) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5486..5503) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(5486..5502) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5495..5517) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 5647..6102 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5734..5753) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5944..5965 /label=F1ori-F /note="F1 origin, forward primer" promoter 6131..6235 /gene="bla" /label=AmpR promoter CDS 6236..7096 /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(6454..6473) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7267..7855 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7756..7775 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 8009..8026 /label=L4440 /note="L4440 vector, forward primer" protein_bind 8143..8164 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 8179..8209 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8217..8233 /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 8222..8244 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 8241..8257 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 8241..8257 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 8276..8296 /label=T3 /note="T3 promoter, forward primer" promoter 8278..8296 /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 ctcgatacaa taaacgggtc 241 tctctggtta gaccagatct gagcctggct ctctggctaa ctagggaacc cactgcttaa 301 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc 361 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc 421 ccgaacaggg acctgaaagc gaaagggaaa ccagagctct ctcgacgcag gactcggctt 481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg 541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga 601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta 661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta 721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga 781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg 841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt 901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga 961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc 1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc 1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct 1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag 1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca 1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg 1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa 1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa 1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga 1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa 1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat 1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt 1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg 1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac tgcgtgcgcc 1801 aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg 1861 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat 1921 tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc 1981 cagtttggtt aattaacccg tgtcggctcc agatctggcc tccgcgccgg gttttggcgc 2041 ctcccgcggg cgcccccctc ctcacggcga gcgctgccac gtcagacgaa gggcgcagcg 2101 agcgtcctga tccttccgcc cggacgctca ggacagcggc ccgctgctca taagactcgg 2161 ccttagaacc ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg 2221 cactggtttt ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt 2281 ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg 2341 tggcacagct agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct 2401 gtgatcgtca cttggtgagt agcgggctgc tgggctggcc ggggctttcg tggccgccgg 2461 gccgctcggt gggacggaag cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg 2521 agcaaggttg ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga 2581 gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc 2641 atggtgggcg gcaagaaccc aaggtcttga ggccttcgct aatgcgggaa agctcttatt 2701 cgggtgagat gggctggggc accatctggg gaccctgacg tgaagtttgt cactgactgg 2761 agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc 2821 acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct 2881 tataatgcag ggtggggcca cctgccggta ggtgtgcggt aggcttttct ccgtcgcagg 2941 acgcagggtt cgggcctagg gtaggctctc ctgaatcgac aggcgccgga cctctggtga 3001 ggggagggat aagtgaggcg tcagtttctt tggtcggttt tatgtaccta tcttcttaag 3061 tagctgaagc tccggttttg aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt 3121 ttttaggcac cttttgaaat gtaatcattt gggtcaatat gtaattttca gtgttagact 3181 agtaaattgt ccgctaaatt ctggccgttt ttggcttttt tgttagacga agcttgggct 3241 gcaggtccga tccaccggtg gatcgatccg atggtgagca agggcgagga ggtcatcaaa 3301 gagttcatgc gcttcaaggt gcgcatggag ggctccatga acggccacga gttcgagatc 3361 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc 3421 aagggcggcc ccctgccctt cgcctgggac atcctgtccc cccagttcat gtacggctcc 3481 aaggcgtacg tgaagcaccc cgccgacatc cccgattaca agaagctgtc cttccccgag 3541 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gtctggtgac cgtgacccag 3601 gactcctccc tgcaggacgg cacgctgatc tacaaggtga agatgcgcgg caccaacttc 3661 ccccccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc caccgagcgc 3721 ctgtaccccc gcgacggcgt gctgaagggc gagatccacc aggccctgaa gctgaaggac 3781 ggcggccact acctggtgga gttcaagacc atctacatgg ccaagaagcc cgtgcaactg 3841 cccggctact actacgtgga caccaagctg gacatcacct cccacaacga ggactacacc 3901 atcgtggaac agtacgagcg ctccgagggc cgccaccacc tgttcctgta cggcatggac 3961 gagctgtaca agggatctgg agcaacaaac ttctcactac tcaaacaagc aggtgacgtg 4021 gaggagaatc ccgggccttc tagaggatcc ggctccggcg agggcagggg aagtctacta 4081 acatgcgggg acgtggagga aaatcccggc ccagctagcg tgtgatcaga attcgttccg 4141 gagtcgtcga ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat 4201 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca 4261 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc 4321 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc 4381 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt 4441 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg 4501 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc 4561 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta 4621 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg 4681 gcctcttccg catcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc 4741 cccgcctgga attaattcga gctcggtacc tttaagacca atgacttaca aggcagctgt 4801 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctacgta actcccaacg 4861 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg 4921 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt 4981 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc 5041 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta 5101 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta 5161 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 5221 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct 5281 atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg 5341 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag 5401 ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct 5461 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa 5521 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta 5581 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat 5641 ggcgcgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg 5701 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc 5761 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc 5821 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta 5881 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta 5941 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg 6001 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa 6061 aatttaacgc gaattttaac aaaatattaa cgtttacaat ttcccaggtg gcacttttcg 6121 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc 6181 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag 6241 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt 6301 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt 6361 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga 6421 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat 6481 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga 6541 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag 6601 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg 6661 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg 6721 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt 6781 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg 6841 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc 6901 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg 6961 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac 7021 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact 7081 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa 7141 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa 7201 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg 7261 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc 7321 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac 7381 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca 7441 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt 7501 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc 7561 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg 7621 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc 7681 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac 7741 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct 7801 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc 7861 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt 7921 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac 7981 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg 8041 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga 8101 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac 8161 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt 8221 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat 8281 taaccctcac taaagggaac aaaagctgga gctgca //