LOCUS V044659 9425 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044659 VERSION V044659 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 2317..3510 /label="IL1R2(NM_004633)" /note="IL1R2(NM_004633)" /gene="IL1R2" CDS 3517..3540 /codon_start=1 /product="8xHis affinity tag" /transl_table=1 /translation="HHHHHHHH" /label="8xHis" promoter 3584..3795 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3808..4076 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4105..4114 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4135..4797 /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 4867..4920 /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 4921..5520 /codon_start=1 /gene="pac from Streptomyces alboniger" /note="confers resistance to puromycin" /product="puromycin N-acetyltransferase" /transl_table=1 /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*" /label="PuroR" misc_feature 5521..6108 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6182..6415 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6487..6608 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6627..6762 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6800..6816) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6824..6840 /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(6848..6878) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6893..6914 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7202..7790) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7961..8821) /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(8822..8926) /gene="bla" /label="AmpR promoter" primer_bind 9400..9416 /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 ctgatacgaa ctcggaattc gccaccatgt tgcgcttgta cgtgttggta 2341 atgggagttt ctgccttcac ccttcagcct gcggcacaca caggggctgc cagaagctgc 2401 cggtttcgtg ggaggcatta caagcgggag ttcaggctgg aaggggagcc tgtagccctg 2461 aggtgccccc aggtgcccta ctggttgtgg gcctctgtca gcccccgcat caacctgaca 2521 tggcataaaa atgactctgc taggacggtc ccaggagaag aagagacacg gatgtgggcc 2581 caggacggtg ctctgtggct tctgccagcc ttgcaggagg actctggcac ctacgtctgc 2641 actactagaa atgcttctta ctgtgacaaa atgtccattg agctcagagt ttttgagaat 2701 acagatgctt tcctgccgtt catctcatac ccgcaaattt taaccttgtc aacctctggg 2761 gtattagtat gccctgacct gagtgaattc acccgtgaca aaactgacgt gaagattcaa 2821 tggtacaagg attctcttct tttggataaa gacaatgaga aatttctaag tgtgaggggg 2881 accactcact tactcgtaca cgatgtggcc ctggaagatg ctggctatta ccgctgtgtc 2941 ctgacatttg cccatgaagg ccagcaatac aacatcacta ggagtattga gctacgcatc 3001 aagaaaaaaa aagaagagac cattcctgtg atcatttccc ccctcaagac catatcagct 3061 tctctggggt caagactgac aatcccgtgt aaggtgtttc tgggaaccgg cacaccctta 3121 accaccatgc tgtggtggac ggccaatgac acccacatag agagcgccta cccgggaggc 3181 cgcgtgaccg aggggccacg ccaggaatat tcagaaaata atgagaacta cattgaagtg 3241 ccattgattt ttgatcctgt cacaagagag gatttgcaca tggattttaa atgtgttgtc 3301 cataataccc tgagttttca gacactacgc accacagtca aggaagcctc ctccacgttc 3361 tcctggggca ttgtgctggc cccactttca ctggccttct tggttttggg gggaatatgg 3421 atgcacagac ggtgcaaaca cagaactgga aaagcagatg gtctgactgt gctatggcct 3481 catcatcaag actttcaatc ctatcccaag ggatcccacc atcaccatca ccatcaccat 3541 tagcggccgc gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg 3601 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg 3661 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt 3721 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt 3781 gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc 3841 cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt 3901 gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt 3961 tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc 4021 ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag 4081 ctgtgaccgg cgcctacgct agacgccacc atggagagcg acgagagcgg cctgcccgcc 4141 atggagatcg agtgccgcat caccggcacc ctgaacggcg tggagttcga gctggtgggc 4201 ggcggagagg gcacccccaa gcagggccgc atgaccaaca agatgaagag caccaaaggc 4261 gccctgacct tcagccccta cctgctgagc cacgtgatgg gctacggctt ctaccacttc 4321 ggcacctacc ccagcggcta cgagaacccc ttcctgcacg ccatcaacaa cggcggctac 4381 accaacaccc gcatcgagaa gtacgaggac ggcggcgtgc tgcacgtgag cttcagctac 4441 cgctacgagg ccggccgcgt gatcggcgac ttcaaggtgg tgggcaccgg cttccccgag 4501 gacagcgtga tcttcaccga caagatcatc cgcagcaacg ccaccgtgga gcacctgcac 4561 cccatgggcg ataacgtgct ggtgggcagc ttcgcccgca ccttcagcct gcgcgacggc 4621 ggctactaca gcttcgtggt ggacagccac atgcacttca agagcgccat ccaccccagc 4681 atcctgcaga acgggggccc catgttcgcc ttccgccgcg tggaggagct gcacagcaac 4741 accgagctgg gcatcgtgga gtaccagcac gccttcaaga cccccatcgc cttcgccaga 4801 tcccgcgctc agtcgtccaa ttctgccgtg gacggcaccg ccggacccgg ctccaccgga 4861 tctcgcgagg gcagaggaag tcttctaaca tgcggtgacg tggaggagaa tcccggccct 4921 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta 4981 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac 5041 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac 5101 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag 5161 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt 5221 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag 5281 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc 5341 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg 5401 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc 5461 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga 5521 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct 5581 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt 5641 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg 5701 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact 5761 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct 5821 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg 5881 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc 5941 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc 6001 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctc 6061 cgccttcgcc ctcagacgag tcggatctcc ctttggccgc ctccccgcct ggtaccttta 6121 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga 6181 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct 6241 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 6301 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 6361 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt 6421 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt 6481 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 6541 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 6601 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt 6661 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct 6721 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg 6781 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 6841 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 6901 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 6961 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 7021 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 7081 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 7141 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 7201 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 7261 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 7321 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 7381 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 7441 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 7501 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 7561 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 7621 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 7681 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 7741 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 7801 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 7861 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 7921 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 7981 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 8041 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 8101 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 8161 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 8221 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 8281 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 8341 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 8401 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 8461 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 8521 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 8581 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 8641 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 8701 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 8761 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 8821 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 8881 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 8941 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 9001 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 9061 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 9121 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 9181 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga 9241 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat 9301 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat 9361 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca 9421 agctg //