LOCUS V021833 9164 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V021833 VERSION V021833 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 1804..1921 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2011..2214 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2303..3043 /label="Gzmb(NM_013542)" /note="Gzmb(NM_013542)" /gene="Gzmb" CDS 3044..3073 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /transl_table=1 /translation="EQKLISEEDL" /label="Myc" promoter 3117..3328 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3341..3609 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3638..3647 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3668..4330 /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" CDS 4400..4453 /codon_start=1 /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /product="2A peptide from Thosea asigna virus capsid protein" /protein_id="" /transl_table=1 /translation="EGRGSLLTCGDVEENPGP" /label="T2A" CDS 4460..5254 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" misc_feature 5261..5849 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5921..6154 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6226..6347 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6366..6501 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6539..6555) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6563..6579 /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(6587..6617) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6632..6653 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6941..7529) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7700..8560) /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(8561..8665) /gene="bla" /label="AmpR promoter" primer_bind 9139..9155 /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 ggtatcggtt 1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat 1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt 1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac 1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg 2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg 2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca 2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta 2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc 2281 tagagctagc gaattcgcca ccatgaagat cctcctgcta ctgctgacct tgtctctggc 2341 ctccaggaca aaggcagggg agatcatcgg gggacatgaa gtcaagcccc actctcgacc 2401 ctacatggcc ttactttcga tcaaggatca gcagcctgag gcgatatgtg ggggcttcct 2461 tattcgagag gactttgtgc tgactgctgc tcactgtgaa ggaagtataa taaatgtcac 2521 tttgggggcc cacaacatca aagaacagga gaagacccag caagtcatcc ctatggtaaa 2581 atgcattccc cacccagact ataatcctaa gacattctcc aatgacatca tgctgctaaa 2641 gctgaagagt aaggccaaga ggactagagc tgtgaggccc ctcaacctgc ccaggcgcaa 2701 tgtcaatgtg aagccaggag atgtgtgcta tgtggctggt tggggaagga tggccccaat 2761 gggcaaatac tcaaacacgc tacaagaggt tgagctgaca gtacagaagg atcgggagtg 2821 tgagtcctac tttaaaaatc gttacaacaa aaccaatcag atatgtgcgg gggacccaaa 2881 gaccaaacgt gcttcctttc ggggggattc tggaggcccg cttgtgtgta aaaaagtggc 2941 tgcaggcata gtttcctatg gatataagga tggttcacct ccacgtgctt tcaccaaagt 3001 ctcgagtttc ttatcctgga taaagaaaac aatgaaaagc agcgagcaaa agctcatttc 3061 tgaagaggac ttgtagcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc 3121 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg 3181 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct 3241 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt 3301 tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc 3361 acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc 3421 tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc 3481 gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac 3541 gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg 3601 ttacagatcc aagctgtgac cggcgcctac gctagacgcc accatggaga gcgacgagag 3661 cggcctgccc gccatggaga tcgagtgccg catcaccggc accctgaacg gcgtggagtt 3721 cgagctggtg ggcggcggag agggcacccc caagcagggc cgcatgacca acaagatgaa 3781 gagcaccaaa ggcgccctga ccttcagccc ctacctgctg agccacgtga tgggctacgg 3841 cttctaccac ttcggcacct accccagcgg ctacgagaac cccttcctgc acgccatcaa 3901 caacggcggc tacaccaaca cccgcatcga gaagtacgag gacggcggcg tgctgcacgt 3961 gagcttcagc taccgctacg aggccggccg cgtgatcggc gacttcaagg tggtgggcac 4021 cggcttcccc gaggacagcg tgatcttcac cgacaagatc atccgcagca acgccaccgt 4081 ggagcacctg caccccatgg gcgataacgt gctggtgggc agcttcgccc gcaccttcag 4141 cctgcgcgac ggcggctact acagcttcgt ggtggacagc cacatgcact tcaagagcgc 4201 catccacccc agcatcctgc agaacggggg ccccatgttc gccttccgcc gcgtggagga 4261 gctgcacagc aacaccgagc tgggcatcgt ggagtaccag cacgccttca agacccccat 4321 cgccttcgcc agatcccgcg ctcagtcgtc caattctgcc gtggacggca ccgccggacc 4381 cggctccacc ggatctcgcg agggcagagg aagtcttcta acatgcggtg acgtggagga 4441 gaatcccggc cctgccacca tggctgaaca agatggattg cacgcaggtt ctccggccgc 4501 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc 4561 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc 4621 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg 4681 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt 4741 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc 4801 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga 4861 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga 4921 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct 4981 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc 5041 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt 5101 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg 5161 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat 5221 cgccttctat cgccttcttg acgagttctt ctgagtcgac aatcaacctc tggattacaa 5281 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata 5341 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc 5401 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg 5461 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac 5521 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat 5581 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt 5641 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat 5701 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc 5761 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag 5821 tcggatctcc ctttgggccg cctccccgcg gtacctttaa gaccaatgac ttacaaggca 5881 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc 5941 caacgaaaat aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga 6001 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 6061 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 6121 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt 6181 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca 6241 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 6301 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc 6361 tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 6421 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt 6481 gaggaggctt ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt 6541 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 6601 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 6661 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 6721 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 6781 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 6841 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 6901 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 6961 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 7021 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 7081 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 7141 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 7201 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 7261 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 7321 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 7381 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 7441 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 7501 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 7561 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 7621 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 7681 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 7741 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 7801 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 7861 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 7921 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 7981 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 8041 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 8101 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 8161 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 8221 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 8281 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 8341 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 8401 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 8461 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 8521 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 8581 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 8641 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag 8701 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc 8761 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 8821 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 8881 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 8941 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 9001 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 9061 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 9121 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctg //