LOCUS V029335 9226 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029335 VERSION V029335 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..3252 /label="Hsd17b12(NM_019657)" /note="Hsd17b12(NM_019657)" /gene="Hsd17b12" CDS 3259..3324 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3368..3579 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3592..3860 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3889..3898 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3919..4581 /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 4651..4704 /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 4705..5304 /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 5305..5893 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5967..6200 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6272..6393 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6433..6568 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6601..6617) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6625..6641 /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(6649..6679) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6694..6715 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7003..7591) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7762..8622) /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(8623..8727) /gene="bla" /label="AmpR promoter" primer_bind 9201..9217 /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 agtgcgctcc cccggcggcc 2341 ggcttcctgt actgggtggg cgcgagcacc attgcctacc tggccctgcg agcctcgtac 2401 tcgctcttca gggccttcca ggtgtggtgc gtgggcaacg aggccttggt cggtccgcga 2461 ctcggagaat gggcagttgt tacaggtggc actgatggaa ttggaaaagc atatgcagaa 2521 gagttagcaa aacgtggaat gaagattgtc ctgatcagca ggtcccaaga taaactgaat 2581 caggtttcca acaacatcaa agaaaaattc aacgttgaaa caaggaccat tgctgtcgac 2641 ttttcattgg atgatattta tgataaaatt aagacaggcc tgtctggtct tgagattggc 2701 gttttagtga acaacgtggg catgtcatac gagtatccgg aatactttct ggaaattccc 2761 gacttggaca acaccatcaa gaaactgata aatattaatg tgctttccgt ttgcaaggtg 2821 acacgcttgg tgctgcctgg catggtagaa agatctaaag gggtgattct caacatctcc 2881 tcagccagtg gcatgctccc agttccattg ttgacaatct actctgcaac caaggccttt 2941 gtagatttct tctctcagtg cctccatgag gagtataaga gcaagggcat ctttgtgcag 3001 agtgtcatgc cataccttgt agctacaaaa ctggcaaaaa tacagaagcc gactttggat 3061 aagccctctg cagagacatt tgtgaagtct gcaattaaaa cagtaggttt gcagacccga 3121 accactggat atgtgatcca ctctctcatg ggctcaataa actcaatcat gcctcgttgg 3181 atgtatttta aaataatcat gggtttcagc aagtctttgc ggaatcgcta cctgaagaaa 3241 aggaagaaga acggatccga ctacaaagac catgacggtg attataaaga tcatgacatc 3301 gactacaagg atgacgatga caagtagcgg ccgcgaagga tctgcgatcg ctccggtgcc 3361 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc 3421 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac 3481 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg 3541 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc 3601 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc 3661 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta 3721 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc 3781 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct 3841 gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagacgc caccatggag 3901 agcgacgaga gcggcctgcc cgccatggag atcgagtgcc gcatcaccgg caccctgaac 3961 ggcgtggagt tcgagctggt gggcggcgga gagggcaccc ccaagcaggg ccgcatgacc 4021 aacaagatga agagcaccaa aggcgccctg accttcagcc cctacctgct gagccacgtg 4081 atgggctacg gcttctacca cttcggcacc taccccagcg gctacgagaa ccccttcctg 4141 cacgccatca acaacggcgg ctacaccaac acccgcatcg agaagtacga ggacggcggc 4201 gtgctgcacg tgagcttcag ctaccgctac gaggccggcc gcgtgatcgg cgacttcaag 4261 gtggtgggca ccggcttccc cgaggacagc gtgatcttca ccgacaagat catccgcagc 4321 aacgccaccg tggagcacct gcaccccatg ggcgataacg tgctggtggg cagcttcgcc 4381 cgcaccttca gcctgcgcga cggcggctac tacagcttcg tggtggacag ccacatgcac 4441 ttcaagagcg ccatccaccc cagcatcctg cagaacgggg gccccatgtt cgccttccgc 4501 cgcgtggagg agctgcacag caacaccgag ctgggcatcg tggagtacca gcacgccttc 4561 aagaccccca tcgccttcgc cagatcccgc gctcagtcgt ccaattctgc cgtggacggc 4621 accgccggac ccggctccac cggatctcgc gagggcagag gaagtcttct aacatgcggt 4681 gacgtggagg agaatcccgg ccctatgacc gagtacaagc ccacggtgcg cctcgccacc 4741 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc 4801 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc 4861 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg 4921 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc 4981 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc 5041 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc 5101 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag 5161 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag 5221 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc 5281 atgacccgca agcccggtgc ctgaaatcaa cctctggatt acaaaatttg tgaaagattg 5341 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct 5401 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg 5461 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact 5521 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc 5581 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc 5641 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa 5701 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc 5761 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg 5821 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg 5881 gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg tagatcttag 5941 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagataaga 6001 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc 6061 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt 6121 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc 6181 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact 6241 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta 6301 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 6361 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc 6421 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 6481 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag 6541 aagtagtgag gaggcttttt tggaggccta gacttttgca gagaccaaat tcgtaatcat 6601 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 6661 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 6721 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 6781 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 6841 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 6901 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 6961 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 7021 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 7081 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 7141 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 7201 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 7261 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 7321 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 7381 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 7441 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 7501 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 7561 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 7621 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 7681 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 7741 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 7801 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 7861 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 7921 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 7981 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 8041 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 8101 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 8161 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 8221 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 8281 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 8341 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 8401 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 8461 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 8521 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 8581 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 8641 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 8701 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta 8761 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg 8821 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt 8881 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg 8941 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt 9001 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg 9061 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 9121 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 9181 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg //