LOCUS Exported 8327 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION 3rd generation Lentiviral vector for bi-cistronic expression of EGFP and the gene of interest. ACCESSION . VERSION . KEYWORDS pUltra SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8327) AUTHORS Lou E, Fujisawa S, Morozov A, Barlas A, Romin Y, Dogan Y, Gholami S, Moreira AL, Manova-Todorova K, Moore MA TITLE Tunneling nanotubes provide a unique conduit for intercellular transfer of cellular contents in human malignant pleural mesothelioma. JOURNAL PLoS One. 2012;7(3):e33093. doi: 10.1371/journal.pone.0033093. Epub 2012 Mar 9. PUBMED 22427958 REFERENCE 2 (bases 1 to 8327) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8327 /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" regulatory 3261..3270 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3267..3983 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 3267..3983 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3312..3333) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3573..3592) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3920..3941 /label=EGFP-C /note="EGFP, forward primer" CDS 3993..4049 /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 4071..4124 /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 4168..4756 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4221..4241) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4639..4650) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 4847..5080 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 5152..5273 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5189..5208) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5243..5262 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 5313..5448 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5375..5394 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(5468..5487) /label=T7 /note="T7 promoter, forward primer" promoter complement(5469..5487) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5497..5514) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(5497..5513) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5506..5528) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 5658..6113 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5745..5764) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5955..5976 /label=F1ori-F /note="F1 origin, forward primer" promoter 6142..6246 /gene="bla" /label=AmpR promoter CDS 6247..7107 /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(6465..6484) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7278..7866 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7767..7786 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 8020..8037 /label=L4440 /note="L4440 vector, forward primer" protein_bind 8154..8175 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 8190..8220 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8228..8244 /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 8233..8255 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 8252..8268 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 8252..8268 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 8287..8307 /label=T3 /note="T3 promoter, forward primer" promoter 8289..8307 /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 tccaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg 3301 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg 3361 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg 3421 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct 3481 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag 3541 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg 3601 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca 3661 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct 3721 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca 3781 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg 3841 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc 3901 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc 3961 tcggcatgga cgagctgtac aagggatctg gagcaacaaa cttctcacta ctcaaacaag 4021 caggtgacgt ggaggagaat cccgggcctt ctagaggatc cggctccggc gagggcaggg 4081 gaagtctact aacatgcggg gacgtggagg aaaatcccgg cccagctagc gtgtgatcag 4141 aattcgttcc ggagtcgtcg actcgacaat caacctctgg attacaaaat ttgtgaaaga 4201 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg 4261 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc 4321 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc 4381 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt 4441 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt 4501 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg 4561 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg 4621 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg 4681 ccggctctgc ggcctcttcc gcatcttcgc cttcgccctc agacgagtcg gatctccctt 4741 tgggccgcct ccccgcctgg aattaattcg agctcggtac ctttaagacc aatgacttac 4801 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctacgt 4861 aactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag 4921 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc 4981 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga 5041 tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt 5101 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt 5161 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca 5221 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc 5281 tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg 5341 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct 5401 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gggacgtacc 5461 caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg 5521 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc 5581 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct 5641 gaatggcgaa tggcgcgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt 5701 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt 5761 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc 5821 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga 5881 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc 5941 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt 6001 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct 6061 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa tttcccaggt 6121 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca 6181 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg 6241 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc 6301 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg 6361 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt 6421 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta 6481 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat 6541 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga 6601 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca 6661 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact 6721 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc 6781 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact 6841 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt 6901 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt 6961 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt 7021 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata 7081 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag 7141 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat 7201 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa 7261 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca 7321 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt 7381 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg 7441 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc 7501 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga 7561 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc 7621 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc 7681 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca 7741 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg 7801 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta 7861 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct 7921 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag 7981 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa 8041 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc 8101 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg 8161 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg 8221 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc 8281 aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctgca //