LOCUS Exported 7686 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION self-inactivating lentiviral plasmid for co-expression of your gene of interest and EGFP. ACCESSION . VERSION . KEYWORDS pHIV-EGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7686) AUTHORS Welm BE, Dijkgraaf GJ, Bledau AS, Welm AL, Werb Z TITLE Lentiviral transduction of mammary stem cells for analysis of gene function during development and cancer. JOURNAL Cell Stem Cell. 2008 Jan 10. 2(1):90-102. PUBMED 18371425 REFERENCE 2 (bases 1 to 7686) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7686 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(47..66) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 238..617 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 619..817 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 772..792 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 835..1015 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1062..1187 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1684..1917 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2102..2146 /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 2444..2561 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2624..3802 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 2855..3793 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 3750..3770 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" misc_feature 3848..4434 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4015..4032) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 4242..4261 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 4435..5154 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 4435..5151 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(4480..4501) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4741..4760) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 5088..5109 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(5166..5182) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5167..5183) /label=pBluescriptKS /note="For pBluescript vector" LTR 5692..5872 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" rep_origin complement(5934..6522) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6014..6033) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6693..7553) /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 7316..7335 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7554..7658) /gene="bla" /label=AmpR promoter ORIGIN 1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg 61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt 121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc 181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac 241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat 301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg 361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt 421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag 481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc 541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag 601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt 661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc 721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg 781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct 841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc 1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc 1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa 1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg 1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata 1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc 1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga 1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc 1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca 1501 aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga 1561 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt 1621 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg 1681 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc 1741 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa 1801 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa 1861 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg 1921 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg 1981 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga 2041 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga 2101 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa 2161 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg 2221 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc 2281 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga 2341 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact 2401 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg 2461 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa 2521 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca 2581 gcagagatcc agtttggtta gtaccgggcc cgctctagcg tgaggctccg gtgcccgtca 2641 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg 2701 aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct 2761 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt 2821 tctttttcgc aacgggtttg ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg 2881 gcctggcctc tttacgggtt atggcccttg cgtgccttga attacttcca cctggctgca 2941 gtacgtgatt cttgatcccg agcttcgggt tggaagtggg tgggagagtt cgaggccttg 3001 cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc gctggggccg 3061 ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct gctttcgata agtctctagc 3121 catttaaaat ttttgatgac ctgctgcgac gctttttttc tggcaagata gtcttgtaaa 3181 tgcgggccaa gatctgcaca ctggtatttc ggtttttggg gccgcgggcg gcgacggggc 3241 ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg cgagcgcggc caccgagaat 3301 cggacggggg tagtctcaag ctggccggcc tgctctggtg cctggcctcg cgccgccgtg 3361 tatcgccccg ccctgggcgg caaggctggc ccggtcggca ccagttgcgt gagcggaaag 3421 atggccgctt cccggccctg ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga 3481 gcgggcgggt gagtcaccca cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc 3541 atgtgactcc acggagtacc gggcgccgtc caggcacctc gattagttct cgagcttttg 3601 gagtacgtcg tctttaggtt ggggggaggg gttttatgcg atggagtttc cccacactga 3661 gtgggtggag actgaagtta ggccagcttg gcacttgatg taattctcct tggaatttgc 3721 cctttttgag tttggatctt ggttcattct caagcctcag acagtggttc aaagtttttt 3781 tcttccattt caggtgtcgt gagcggccgc tgagttaact attctagagt acccgggcta 3841 ggatccgccc ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag 3901 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga 3961 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg 4021 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt 4081 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca 4141 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc 4201 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat 4261 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc 4321 ctcggtacac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga 4381 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatggtg 4441 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac 4501 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag 4561 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg 4621 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac 4681 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag 4741 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac 4801 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg 4861 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc 4921 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac 4981 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg 5041 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg 5101 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc 5161 cgcatcgata ccgtcgacct cgatcgagac ctagaaaaac atggagcaat cacaagtagc 5221 aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg 5281 ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat 5341 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga 5401 caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga ttggcagaac 5461 tacacaccag ggccagggat cagatatcca ctgacctttg gatggtgcta caagctagta 5521 ccagttgagc aagagaaggt agaagaagcc aatgaaggag agaacacccg cttgttacac 5581 cctgtgagcc tgcatgggat ggatgacccg gagagagaag tattagagtg gaggtttgac 5641 agccgcctag catttcatca catggcccga gagctgcatc cggactgtac tgggtctctc 5701 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag 5761 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct 5821 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagcatgtga 5881 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 5941 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 6001 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 6061 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 6121 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 6181 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 6241 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 6301 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 6361 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 6421 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 6481 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 6541 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 6601 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 6661 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 6721 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 6781 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 6841 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 6901 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 6961 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 7021 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 7081 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 7141 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 7201 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 7261 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 7321 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 7381 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 7441 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 7501 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 7561 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 7621 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 7681 cctgac //