LOCUS V038114 8746 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V038114 VERSION V038114 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..2772 /label="Srgn(NM_011157)" /note="Srgn(NM_011157)" /gene="Srgn" CDS 2779..2844 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 2888..3099 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3112..3380 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3409..3418 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3439..4101 /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 4171..4224 /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 4225..4824 /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 4825..5413 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5487..5720 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5792..5913 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5953..6088 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6121..6137) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6145..6161 /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(6169..6199) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6214..6235 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6523..7111) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7282..8142) /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(8143..8247) /gene="bla" /label="AmpR promoter" primer_bind 8721..8737 /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 gccaccatgc aggttcccgt cggcagcagg 2341 cttgtcctgg ctctcgcctt cgtcctggtt tggggatctt cagtgcaagg ttatcctgct 2401 cggagagcca ggtaccagtg ggtccgctgc aaaccgaatg gcttttttgc gaactgcatc 2461 gaggagaagg gaccacagtt tgacctaata gatgaatcca ataacatcgg ccctcccatg 2521 aataatcctg ttttgatgga aggaccctca aaagatttca tctccaatta tgatgactat 2581 gggtcaggtt cgggctccgg ctctggctcc ggctctggct cgggttccgg ctccggaagt 2641 ggcttcctag gtgacatgga atgggaatac cagccaacag atgaaagcaa tattgtctat 2701 ttcaactata agccttttga caggattctc actgagcaaa accaagacca accagaagac 2761 gattttatta taggatccga ctacaaagac catgacggtg attataaaga tcatgacatc 2821 gactacaagg atgacgatga caagtagcgg ccgcgaagga tctgcgatcg ctccggtgcc 2881 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc 2941 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac 3001 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg 3061 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc 3121 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc 3181 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta 3241 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc 3301 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct 3361 gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagacgc caccatggag 3421 agcgacgaga gcggcctgcc cgccatggag atcgagtgcc gcatcaccgg caccctgaac 3481 ggcgtggagt tcgagctggt gggcggcgga gagggcaccc ccaagcaggg ccgcatgacc 3541 aacaagatga agagcaccaa aggcgccctg accttcagcc cctacctgct gagccacgtg 3601 atgggctacg gcttctacca cttcggcacc taccccagcg gctacgagaa ccccttcctg 3661 cacgccatca acaacggcgg ctacaccaac acccgcatcg agaagtacga ggacggcggc 3721 gtgctgcacg tgagcttcag ctaccgctac gaggccggcc gcgtgatcgg cgacttcaag 3781 gtggtgggca ccggcttccc cgaggacagc gtgatcttca ccgacaagat catccgcagc 3841 aacgccaccg tggagcacct gcaccccatg ggcgataacg tgctggtggg cagcttcgcc 3901 cgcaccttca gcctgcgcga cggcggctac tacagcttcg tggtggacag ccacatgcac 3961 ttcaagagcg ccatccaccc cagcatcctg cagaacgggg gccccatgtt cgccttccgc 4021 cgcgtggagg agctgcacag caacaccgag ctgggcatcg tggagtacca gcacgccttc 4081 aagaccccca tcgccttcgc cagatcccgc gctcagtcgt ccaattctgc cgtggacggc 4141 accgccggac ccggctccac cggatctcgc gagggcagag gaagtcttct aacatgcggt 4201 gacgtggagg agaatcccgg ccctatgacc gagtacaagc ccacggtgcg cctcgccacc 4261 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc 4321 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc 4381 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg 4441 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc 4501 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc 4561 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc 4621 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag 4681 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag 4741 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc 4801 atgacccgca agcccggtgc ctgaaatcaa cctctggatt acaaaatttg tgaaagattg 4861 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct 4921 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg 4981 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact 5041 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc 5101 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc 5161 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa 5221 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc 5281 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg 5341 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg 5401 gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg tagatcttag 5461 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagataaga 5521 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc 5581 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt 5641 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc 5701 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact 5761 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta 5821 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 5881 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc 5941 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 6001 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag 6061 aagtagtgag gaggcttttt tggaggccta gacttttgca gagaccaaat tcgtaatcat 6121 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 6181 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 6241 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 6301 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 6361 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 6421 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 6481 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 6541 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 6601 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 6661 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 6721 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 6781 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 6841 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 6901 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 6961 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 7021 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 7081 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 7141 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 7201 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 7261 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 7321 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 7381 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 7441 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 7501 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 7561 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 7621 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 7681 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 7741 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 7801 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 7861 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 7921 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 7981 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 8041 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 8101 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 8161 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 8221 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta 8281 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg 8341 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt 8401 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg 8461 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt 8521 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg 8581 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 8641 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 8701 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg //