LOCUS Exported 9318 bp ds-DNA circular SYN 27-JUL-2023 DEFINITION Expresses chicken cytoplasmic ovalbumin in lentiviral vector with puromycin resistance. ACCESSION . VERSION . KEYWORDS pLVX-puro-cOVA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9318) TITLE pLVX-OVA JOURNAL Unpublished REFERENCE 2 (bases 1 to 9318) AUTHORS . TITLE Direct Submission JOURNAL Exported Jul 27, 2023 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9318 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 1..634 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 681..806 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1303..1536 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1721..1765 /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 2028..2143 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" enhancer 2201..2504 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2505..2716 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2667..2687 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 2713..2737 /label=LNCX /note="Human CMV promoter, forward primer" intron 2877..3009 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 2992..3011 /label=pMT2-F /note="Synthetic intron, forward primer" promoter 3054..3072 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 3488..3511 /codon_start=1 /product="antigenic CD8+ T cell epitope from chicken ovalbumin" /label=OVA peptide /translation="SIINFEKL" promoter complement(3929..3946) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" polyA_signal 3964..4085 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" promoter 4153..4652 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(4188..4211) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 4585..4604 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 4673..5272 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(4673..4692) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 5169..5189 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 5286..5874 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5339..5359) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5757..5768) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 6081..6714 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" primer_bind complement(6843..6859) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6843..6859) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6856..6878) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6867..6883 /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)." promoter complement(6891..6921) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6936..6957 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7074..7091) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7245..7833) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7325..7344) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8004..8864) /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 8627..8646 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8865..8969) /gene="bla" /label=AmpR promoter polyA_signal 9017..9151 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag 2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa 2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg 2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt 2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtccgccc cctattgacg 2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta cgggactttc 2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc 2521 agtacaccaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca 2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta 2641 acaactgcga tcgcccgccc cgttgacgca aatgggcggt aggcgtgtac ggtgggaggt 2701 ctatataagc agagctcgtt tagtgaaccg tcagatcact agaagcttta ttgcggtagt 2761 ttatcacagt taaattgcta acgcagtcag tgcttctgac acaacagtct cgaacttaag 2821 ctgcagtgac tctcttaagg tagccttgca gaagttggtc gtgaggcact gggcaggtaa 2881 gtatcaaggt tacaagacag gtttaaggag accaatagaa actgggcttg tcgagacaga 2941 gaagactctt gcgtttctga taggcaccta ttggtcttac tgacatccac tttgcctttc 3001 tctccacagg tgtccactcc cagttcaatt acagctctta aggctagagt acttaatacg 3061 actcactata ggctagcctc gagaattcgt cgacatgggc tccatcggcg cagcaagcat 3121 ggaattttgt tttgatgtat tcaaggagct catcaattcc tgggtagaaa gtcagacaaa 3181 tggaattatc agaaatgtcc ttcagccaag ctccgtggat tctcaaactg caatggttct 3241 ggttaatgcc attgtcttca aaggactgtg ggagaaaaca tttaaggatg aagacacaca 3301 agcaatgcct ttcagagtga ctgagcaaga aagcaaacct gtgcagatga tgtaccagat 3361 tggtttattt agagtggcat caatggcttc tgagaaaatg aagatcctgg agcttccatt 3421 tgccagtggg acaatgagca tgttggtgct gttgcctgat gaagtctcag gccttgagca 3481 gcttgagagt ataatcaact ttgaaaaact gactgaatgg accagttcta atgttatgga 3541 agagaggaag atcaaagtgt acttacctcg catgaagatg gaggaaaaat acaacctcac 3601 atctgtctta atggctatgg gcattactga cgtgtttagc tcttcagcca atctgtctgg 3661 catctcctca gcagagagcc tgaagatatc tcaagctgtc catgcagcac atgcagaaat 3721 caatgaagca ggcagagagg tggtagggtc agcagaggct ggagtggatg ctgcaagcgt 3781 ctctgaagaa tttagggctg accatccatt cctcttctgt atcaagcaca tcgcaaccaa 3841 cgccgttctc ttctttggca gatgtgtttc cccttaagtc gacgaattca cgcgtggtac 3901 ctctagagtc gacccgggcg gccgcttccc tttagtgagg gttaatgctt cgagcagaca 3961 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct 4021 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac 4081 aagttaacaa caacaattct gcagtcgacg gtaccgcggg cccgggatcc cgcgactcta 4141 gataattcta ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag 4201 cagccccgct gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat 4261 ccaccggtag gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc 4321 ctcccctagt caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa 4381 tggaagtagc acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc 4441 gggtaggcct ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga 4501 ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg 4561 cccgaaggtc ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc 4621 tgttctcctc ttcctcatct ccgggccttt cgacctgcag cccaagctta ccatgaccga 4681 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct 4741 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat 4801 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa 4861 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga 4921 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct 4981 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg 5041 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt 5101 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc 5161 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga 5221 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaccgcgtct 5281 ggaacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg 5341 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt 5401 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg 5461 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc 5521 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc 5581 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc 5641 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc 5701 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg 5761 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc 5821 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa 5881 ttaattctgc agtcgagacc tagaaaaaca tggagcaatc acaagtagca atacagcagc 5941 taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt 6001 cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt 6061 tttaaaagaa aagaggggac tggaagggct aattcactcc caacgaagac aagatatcct 6121 tgatctgtgg atctaccaca cacaaggcta cttccctgat tagcagaact acacaccagg 6181 gccaggggtc agatatccac tgacctttgg atggtgctac aagctagtac cagttgagcc 6241 agataaggta gaagaggcca ataaaggaga gaacaccagc ttgttacacc ctgtgagcct 6301 gcatgggatg gatgacccgg agagagaagt gttagagtgg aggtttgaca gccgcctagc 6361 atttcatcac gtggcccgag agctgcatcc ggagtacttc aagaactgct gatatcgagc 6421 ttgctacaag ggactttccg ctggggactt tccagggagg cgtggcctgg gcgggactgg 6481 ggagtggcga gccctcagat cctgcatata agcagctgct ttttgcctgt actgggtctc 6541 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 6601 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 6661 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta 6721 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg 6781 agaggccttg acattgctag cgttttaccg tcgacctcta gctagagctt ggcgtaatca 6841 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga 6901 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt 6961 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga 7021 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc 7081 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg 7141 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc 7201 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 7261 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 7321 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 7381 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat 7441 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 7501 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 7561 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 7621 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 7681 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 7741 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag 7801 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg 7861 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa 7921 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata 7981 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg 8041 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata 8101 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg 8161 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct 8221 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt 8281 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc 8341 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga 8401 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt 8461 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc 8521 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa 8581 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca 8641 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca 8701 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct 8761 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc 8821 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa 8881 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt 8941 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc 9001 gacggatcgg gagatcaact tgtttattgc agcttataat ggttacaaat aaagcaatag 9061 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 9121 actcatcaat gtatcttatc atgtctggat caactggata actcaagcta accaaaatca 9181 tcccaaactt cccaccccat accctattac cactgccaat tacctgtggt ttcatttact 9241 ctaaacctgt gattcctctg aattattttc attttaaaga aattgtattt gttaaatatg 9301 tactacaaac ttagtagt //