LOCUS Exported 8326 bp ds-DNA circular SYN 16-AUG-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 8326) 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 8326) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 16, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8326 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 94..111 /label=L4440 /note="L4440 vector, forward primer" protein_bind 228..249 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 264..294 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 302..318 /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 307..329 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 326..342 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 326..342 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 361..381 /label=T3 /note="T3 promoter, forward primer" promoter 363..381 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 409..635 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 636..816 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 863..988 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1481..1714 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1899..1943 /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 2241..2358 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2423..3634 /label=UbC promoter /note="human ubiquitin C promoter" primer_bind 3471..3490 /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" regulatory 3667..3676 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3673..4389 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 3673..4389 /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(3718..3739) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3979..3998) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4326..4347 /label=EGFP-C /note="EGFP, forward primer" CDS 4399..4455 /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 4477..4530 /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 4574..5162 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4627..4647) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5045..5056) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 5253..5486 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 5558..5679 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5595..5614) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5649..5668 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 5719..5854 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5781..5800 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(5874..5893) /label=T7 /note="T7 promoter, forward primer" promoter complement(5875..5893) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5903..5920) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(5903..5919) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5912..5934) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 6061..6516 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6148..6167) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6358..6379 /label=F1ori-F /note="F1 origin, forward primer" promoter 6542..6646 /gene="bla" /label=AmpR promoter CDS 6647..7507 /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(6865..6884) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7678..8266 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8167..8186 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct 61 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa 121 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg 181 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag 241 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga 301 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc 361 gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg tagtcttatg 421 caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc ttacaaggag 481 agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt gccttattag 541 gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg cattgcagag 601 atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta gaccagatct 661 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc 721 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc 781 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa 841 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac 901 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta 961 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg 1021 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg 1081 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg 1141 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag 1201 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga 1261 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca 1321 gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg 1381 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa 1441 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt 1501 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca 1561 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc 1621 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg 1681 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac 1741 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga 1801 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa 1861 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg 1921 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata 1981 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac 2041 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc 2101 caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca 2161 gagacagatc cattcgatta gtgaacggat cggcactgcg tgcgccaatt ctgcagacaa 2221 atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg 2281 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt 2341 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt ttggttaatt 2401 aacccgtgtc ggctccagat ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc 2461 cccctcctca cggcgagcgc tgccacgtca gacgaagggc gcagcgagcg tcctgatcct 2521 tccgcccgga cgctcaggac agcggcccgc tgctcataag actcggcctt agaaccccag 2581 tatcagcaga aggacatttt aggacgggac ttgggtgact ctagggcact ggttttcttt 2641 ccagagagcg gaacaggcga ggaaaagtag tcccttctcg gcgattctgc ggagggatct 2701 ccgtggggcg gtgaacgccg atgattatat aaggacgcgc cgggtgtggc acagctagtt 2761 ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg 2821 gtgagtagcg ggctgctggg ctggccgggg ctttcgtggc cgccgggccg ctcggtggga 2881 cggaagcgtg tggagagacc gccaagggct gtagtctggg tccgcgagca aggttgccct 2941 gaactggggg ttggggggag cgcagcaaaa tggcggctgt tcccgagtct tgaatggaag 3001 acgcttgtga ggcgggctgt gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa 3061 gaacccaagg tcttgaggcc ttcgctaatg cgggaaagct cttattcggg tgagatgggc 3121 tggggcacca tctggggacc ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt 3181 cgtctgttgc gggggcggca gttatggcgg tgccgttggg cagtgcaccc gtacctttgg 3241 gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata atgcagggtg 3301 gggccacctg ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc agggttcggg 3361 cctagggtag gctctcctga atcgacaggc gccggacctc tggtgagggg agggataagt 3421 gaggcgtcag tttctttggt cggttttatg tacctatctt cttaagtagc tgaagctccg 3481 gttttgaact atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt aggcaccttt 3541 tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt tagactagta aattgtccgc 3601 taaattctgg ccgtttttgg cttttttgtt agacgaagct tgggctgcag gtccgatcca 3661 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg 3721 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc 3781 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg 3841 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc 3901 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag 3961 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag 4021 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac 4081 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac 4141 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc 4201 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg 4261 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc 4321 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag 4381 ctgtacaagg gatctggagc aacaaacttc tcactactca aacaagcagg tgacgtggag 4441 gagaatcccg ggccttctag aggatccggc tccggcgagg gcaggggaag tctactaaca 4501 tgcggggacg tggaggaaaa tcccggccca gctagcgtgt gatcagaatt cgttccggag 4561 tcgtcgactc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct 4621 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc 4681 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct 4741 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga 4801 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc 4861 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac 4921 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt 4981 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt 5041 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc 5101 tcttccgcat cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc 5161 gcctggaatt aattcgagct cggtaccttt aagaccaatg acttacaagg cagctgtaga 5221 tcttagccac tttttaaaag aaaagggggg actggaaggg ctacgtaact cccaacgaag 5281 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga 5341 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct 5401 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt 5461 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt 5521 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa 5581 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca 5641 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc 5701 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc 5761 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta 5821 ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata 5881 gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc 5941 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata 6001 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg 6061 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg 6121 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca 6181 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta 6241 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc 6301 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg 6361 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat 6421 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta 6481 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt 6541 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag 6601 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca 6661 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc 6721 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat 6781 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc 6841 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg 6901 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc 6961 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat 7021 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga 7081 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc 7141 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc 7201 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt 7261 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc 7321 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc 7381 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca 7441 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca 7501 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt 7561 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta 7621 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg 7681 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc 7741 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag 7801 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa 7861 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc 7921 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc 7981 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta 8041 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag 8101 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct 8161 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctcacttga 8221 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc 8281 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatg //