LOCUS V047371 8710 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V047371 VERSION V047371 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..2736 /label="ATG12(NM_004707)" /note="ATG12(NM_004707)" /gene="ATG12" CDS 2743..2808 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 2852..3063 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3076..3344 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3373..3382 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3403..4065 /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 4135..4188 /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 4189..4788 /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 4789..5377 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5451..5684 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5756..5877 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5917..6052 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6085..6101) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6109..6125 /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(6133..6163) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6178..6199 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6487..7075) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7246..8106) /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(8107..8211) /gene="bla" /label="AmpR promoter" primer_bind 8685..8701 /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 gccaccatgg cggaggagcc gcagtctgtg 2341 ttgcagcttc ctacttcaat tgctgctgga ggggaaggac ttacggatgt ctccccagaa 2401 acaaccaccc cggagccccc gtcttccgct gcagtttccc cgggaacaga ggaacctgct 2461 ggcgacacca agaaaaaaat tgacattttg ctaaaggctg tgggagacac tcctattatg 2521 aaaacaaaga agtgggcagt agagcgaaca cgaaccatcc aaggactcat tgacttcatc 2581 aaaaagtttc ttaaacttgt ggcctcagaa cagttgttta tttatgtgaa tcagtccttt 2641 gctccttccc cagaccaaga agttggaact ctctatgagt gttttggcag tgatggtaaa 2701 ctggttttac attactgcaa gtctcaggcg tggggaggat ccgactacaa agaccatgac 2761 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta gcggccgcga 2821 aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc 2881 gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta 2941 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg 3001 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca 3061 cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc 3121 cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact 3181 gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc 3241 ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa 3301 ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg 3361 cctacgctag acgccaccat ggagagcgac gagagcggcc tgcccgccat ggagatcgag 3421 tgccgcatca ccggcaccct gaacggcgtg gagttcgagc tggtgggcgg cggagagggc 3481 acccccaagc agggccgcat gaccaacaag atgaagagca ccaaaggcgc cctgaccttc 3541 agcccctacc tgctgagcca cgtgatgggc tacggcttct accacttcgg cacctacccc 3601 agcggctacg agaacccctt cctgcacgcc atcaacaacg gcggctacac caacacccgc 3661 atcgagaagt acgaggacgg cggcgtgctg cacgtgagct tcagctaccg ctacgaggcc 3721 ggccgcgtga tcggcgactt caaggtggtg ggcaccggct tccccgagga cagcgtgatc 3781 ttcaccgaca agatcatccg cagcaacgcc accgtggagc acctgcaccc catgggcgat 3841 aacgtgctgg tgggcagctt cgcccgcacc ttcagcctgc gcgacggcgg ctactacagc 3901 ttcgtggtgg acagccacat gcacttcaag agcgccatcc accccagcat cctgcagaac 3961 gggggcccca tgttcgcctt ccgccgcgtg gaggagctgc acagcaacac cgagctgggc 4021 atcgtggagt accagcacgc cttcaagacc cccatcgcct tcgccagatc ccgcgctcag 4081 tcgtccaatt ctgccgtgga cggcaccgcc ggacccggct ccaccggatc tcgcgagggc 4141 agaggaagtc ttctaacatg cggtgacgtg gaggagaatc ccggccctat gaccgagtac 4201 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc 4261 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag 4321 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg 4381 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg 4441 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc 4501 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc 4561 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg 4621 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg 4681 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg 4741 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaaa tcaacctctg 4801 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta 4861 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt 4921 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc 4981 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt 5041 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg 5101 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac 5161 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc 5221 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac 5281 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct 5341 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac 5401 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct 5461 aattcactcc caacgaagat aagatctgct ttttgcttgt actgggtctc tctggttaga 5521 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata 5581 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta 5641 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca 5701 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt 5761 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 5821 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 5881 tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt 5941 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc 6001 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt 6061 tgcagagacc aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct 6121 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 6181 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 6241 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 6301 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 6361 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 6421 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 6481 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 6541 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 6601 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 6661 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 6721 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 6781 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 6841 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 6901 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 6961 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 7021 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 7081 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 7141 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 7201 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 7261 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 7321 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 7381 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 7441 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 7501 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 7561 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 7621 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 7681 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 7741 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 7801 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 7861 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 7921 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 7981 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 8041 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 8101 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 8161 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 8221 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 8281 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 8341 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 8401 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 8461 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt 8521 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg 8581 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 8641 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 8701 tgccaagctg //