LOCUS V042513 9605 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V042513 VERSION V042513 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 234..414 /note="truncated 5' long terminal repeat (LTR) from HIV-1" /label="5' LTR (truncated)" misc_feature 458..583 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1076..1309 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1493..1537 /codon_start=1 /note="recognized by the 2H10 single-chain llama nanobody" /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /transl_table=1 /translation="KNEQELLELDKWASL" /label="gp41 peptide" misc_feature 1798..1915 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2005..2208 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2297..4213 /label="HSPA2(NM_001387931)" /note="HSPA2(NM_001387931)" /gene="HSPA2" CDS 4238..4282 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /transl_table=1 /translation="KETAAAKFERQHMDS" /label="S-Tag" CDS 4316..4339 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" promoter 4408..4619 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4632..4900 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4929..4938 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4959..5621 /codon_start=1 /product="green fluorescent protein 2 from Pontellina plumata, also known as ppluGFP2 (Shagin et al., 2004)" /transl_table=1 /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA" /label="CopGFP" misc_feature 5700..6288 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6362..6595 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6667..6788 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6807..6942 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6980..6996) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7004..7020 /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-β-D-thiogalactopyranoside (IPTG)." /label="lac operator" promoter complement(7028..7058) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7073..7094 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7382..7970) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8141..9001) /codon_start=1 /gene="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /product="β-lactamase" /transl_table=1 /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA SLIKHW*" /label="AmpR" promoter complement(9002..9106) /gene="bla" /label="AmpR promoter" primer_bind 9580..9596 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" ORIGIN 1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca 61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta 121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga 181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc 241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg 421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct 481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt 541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag 601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt 661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt 721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg 781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag 841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag 901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg 961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag 1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag 1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc 1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga 1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc 1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg 1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata 1381 aatctctgga acagattgga 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 ggatctcgac ggttaacttt 1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc 1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga 1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac 1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga 2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg 2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg 2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac 2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc 2281 tagcgaattc gccaccatgt ctgcccgtgg cccggctatc ggcatcgacc tgggcaccac 2341 ctattcgtgc gtcggggtct tccaacatgg caaggtggag atcatcgcca acgaccaggg 2401 caatcgcacc acccccagct acgtggcctt cacggacacc gagcgcctca tcggcgacgc 2461 cgccaagaac caggtggcca tgaaccccac caacaccatc ttcgacgcca agaggctgat 2521 tggacggaaa ttcgaggatg ccacagtgca gtcggatatg aaacactggc cgttccgggt 2581 ggtgagcgag ggaggcaagc ccaaagtgca agtagagtac aagggggaga ccaagacctt 2641 cttcccagag gagatatcct ccatggtcct cacgaagatg aaggagatcg cggaagccta 2701 cctggggggc aaggtgcaca gcgcggtcat aacggtcccg gcctatttca acgactcgca 2761 gcgccaggcc accaaggacg caggcaccat cacggggctc aatgtgctgc gcatcatcaa 2821 cgagcccacg gcggcggcca tcgcctacgg cctggacaag aagggctgcg cgggcggcga 2881 gaagaacgtg ctcatctttg acctgggcgg tggcactttc gacgtgtcca tcctgaccat 2941 cgaggatggc atcttcgagg tgaagtccac ggccggcgac acccacctgg gcggtgagga 3001 cttcgacaac cgcatggtga gccacctggc ggaggagttc aagcgcaagc acaagaagga 3061 cattgggccc aacaagcgcg ccgtgaggcg gctgcgcacc gcttgcgagc gcgccaagcg 3121 caccctgagc tcgtccacgc aggcgagcat cgagatcgac tcgctctacg agggcgtgga 3181 cttctatacg tccatcacgc gcgcccgctt cgaggagctc aatgccgacc tctttcgcgg 3241 gaccctggag ccggtggaga aggcgctgcg cgacgccaag ctggacaagg gccagatcca 3301 ggagatcgtg ctggtgggcg gctccactcg tatccccaag atccagaagc tgctgcagga 3361 tttcttcaac ggcaaggagc tgaacaagag catcaacccc gacgaggcgg tggcctatgg 3421 cgccgcggtg caggcggcca tcctcatcgg cgacaaatca gagaatgtgc aggacctgct 3481 gctactcgac gtgaccccgt tgtcgctggg catcgagaca gctggcggtg tcatgacccc 3541 actcatcaag aggaacacca cgatccccac caagcagacg cagaccttca ccacctactc 3601 ggacaaccag agcagcgtac tggtgcaggt atacgagggc gaacgggcca tgaccaagga 3661 caataacctg ctgggcaagt tcgacctgac cgggattccc cctgcgcctc gcggggtccc 3721 ccaaatcgag gttaccttcg acattgacgc caatggcatc cttaacgtta ccgccgccga 3781 caagagcacc ggtaaggaaa acaaaatcac catcaccaat gacaaaggtc gtctgagcaa 3841 ggacgacatt gaccggatgg tgcaggaggc ggagcggtac aaatcggaag atgaggcgaa 3901 tcgcgaccga gtcgcggcca aaaacgccct ggagtcctat acctacaaca tcaagcagac 3961 ggtggaagac gagaaactga ggggcaagat tagcgagcag gacaaaaaca agatcctcga 4021 caagtgtcag gaggtgatca actggctcga ccgaaaccag atggcagaga aagatgagta 4081 tgaacacaag cagaaagagc tcgaaagagt ttgcaacccc atcatcagca aactttacca 4141 aggtggtcct ggcggcggca gcggcggcgg cggttcagga gcctccgggg gacccaccat 4201 cgaagaagtg gacggctcga gctcaagctt cgaattcaaa gaaaccgctg ctgctaaatt 4261 cgaacgccag cacatggaca gcggagccgg cgcaggagcc ggcgctgacg cgcctgacta 4321 caaagacgat gacgacaagg gagattacaa ggatgactag ggatccgcgg ccgcgaagga 4381 tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga 4441 agttgggggg aggggtcggc aattgaacgg gtgcctagag aaggtggcgc ggggtaaact 4501 gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata 4561 taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc 4621 tgaagcttcg aggggctcgc atctctcctt cacgcgcccg ccgccctacc tgaggccgcc 4681 atccacgccg gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt 4741 ccgccgtcta ggtaagttta aagctcaggt cgagaccggg cctttgtccg gcgctccctt 4801 ggagcctacc tagactcagc cggctctcca cgctttgcct gaccctgctt gctcaactct 4861 acgtctttgt ttcgttttct gttctgcgcc gttacagatc caagctgtga ccggcgccta 4921 cgctagacgc caccatggag agcgacgaga gcggcctgcc cgccatggag atcgagtgcc 4981 gcatcaccgg caccctgaac ggcgtggagt tcgagctggt gggcggcgga gagggcaccc 5041 ccaagcaggg ccgcatgacc aacaagatga agagcaccaa aggcgccctg accttcagcc 5101 cctacctgct gagccacgtg atgggctacg gcttctacca cttcggcacc taccccagcg 5161 gctacgagaa ccccttcctg cacgccatca acaacggcgg ctacaccaac acccgcatcg 5221 agaagtacga ggacggcggc gtgctgcacg tgagcttcag ctaccgctac gaggccggcc 5281 gcgtgatcgg cgacttcaag gtggtgggca ccggcttccc cgaggacagc gtgatcttca 5341 ccgacaagat catccgcagc aacgccaccg tggagcacct gcaccccatg ggcgataacg 5401 tgctggtggg cagcttcgcc cgcaccttca gcctgcgcga cggcggctac tacagcttcg 5461 tggtggacag ccacatgcac ttcaagagcg ccatccaccc cagcatcctg cagaacgggg 5521 gccccatgtt cgccttccgc cgcgtggagg agctgcacag caacaccgag ctgggcatcg 5581 tggagtacca gcacgccttc aagaccccca tcgccttcgc cagatcccgc gctcagtcgt 5641 ccaattctgc cgtggacggc accgccggac ccggctccac cggatctcgc taagtcgaca 5701 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc 5761 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta 5821 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt 5881 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg 5941 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta 6001 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt 6061 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg 6121 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca 6181 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc 6241 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggtaccttta 6301 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga 6361 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct 6421 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 6481 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 6541 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt 6601 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt 6661 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 6721 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 6781 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt 6841 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct 6901 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg 6961 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 7021 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 7081 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 7141 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 7201 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 7261 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 7321 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 7381 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 7441 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 7501 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 7561 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 7621 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 7681 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 7741 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 7801 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 7861 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 7921 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 7981 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 8041 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 8101 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 8161 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 8221 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 8281 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 8341 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 8401 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 8461 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 8521 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 8581 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 8641 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 8701 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 8761 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 8821 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 8881 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 8941 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 9001 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 9061 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 9121 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 9181 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 9241 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 9301 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 9361 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga 9421 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat 9481 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat 9541 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca 9601 agctg //