LOCUS V037255 10006 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V037255 VERSION V037255 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..4032 /label="Me1(NM_008615)" /note="Me1(NM_008615)" /gene="Me1" CDS 4039..4104 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4148..4359 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4372..4640 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4669..4678 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4699..5361 /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 5431..5484 /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 5485..6084 /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 6085..6673 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6747..6980 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7052..7173 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7213..7348 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7381..7397) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7405..7421 /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(7429..7459) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7474..7495 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7783..8371) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8542..9402) /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(9403..9507) /gene="bla" /label="AmpR promoter" primer_bind 9981..9997 /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 agccccgagc cccccgccgc 2341 cgacacaccc accagcgcgg ctacctgctg acgcgggacc cgcatctcaa caaggacttg 2401 gcttttactc tggaagagag acagcagttg aacattcatg gattgttgcc gccctgcatc 2461 atcagccagg agctccaggt ccttagaata attaagaatt tcgaacgact gaactctgac 2521 ttcgacaggt atctcctgtt aatggacctg caagacagaa atgagaagct cttctacagc 2581 gtgctcatgt ctgatgttga aaagttcatg cctattgttt acacccccac cgtgggcctc 2641 gcatgccagc agtacagttt ggcattccgg aagccaagag gcctctttat tagtatccat 2701 gacaaagggc acattgcttc agttcttaat gcatggccag aggatgtcgt caaggctatt 2761 gtggtaactg atggagagcg catccttggc ttgggagacc ttggctgtaa tgggatgggc 2821 atccctgtgg gtaaactggc cctgtacacg gcatgtggag gggtgaaccc acaacagtgt 2881 ctacccatca ctttggatgt gggaacagaa aatgaggagt tacttaagga tccactgtac 2941 atcgggctgc ggcaccggcg agtcagaggc cctgagtatg acgccttcct ggatgagttc 3001 atggaggcag cgtcttccaa atatggcatg aattgcctta ttcagtttga agattttgcc 3061 aatcggaatg catttcgtct cctgaacaag tatcgaaaca agtattgcac atttaacgat 3121 gatattcaag gaacagcgtc tgttgcggtt gcgggtctcc ttgcagctct tcgaataacc 3181 aagaacaagc tctctgatca gacagtgctg ttccagggag ctggagaggc tgccttgggg 3241 attgctcact tggttgttat ggccatggag aaagaaggtt tatcaaagga gaatgctaga 3301 aagaagatat ggttggttga ctcaaaagga ctaatagtta agggtcgtgc atctctcaca 3361 gaagagaaag aggtgtttgc ccatgaacat gaagaaatga agaatctgga agccattgtt 3421 caaaagataa aaccaactgc cctcatagga gttgctgcaa ttggtggtgc tttcactgaa 3481 caaattctca aggatatggc tgccttcaac gagcggccca tcatctttgc tttgagtaat 3541 ccgaccagca aagcggagtg ctctgcagag cagtgctaca aggtgaccaa gggacgtgca 3601 atctttgcca gcggcagtcc ttttgatcca gtcactctcc cagatggacg gactctgttt 3661 cctggccaag gcaacaattc ctacgtgttc cctggagttg ctcttggggt ggtggcctgc 3721 ggactgagac acatcgatga taaggtcttc ctcaccactg ctgaggtcat atctcagcaa 3781 gtgtcagata aacacctgca agaaggccgg ctctatcctc ctttgaatac cattcgaggc 3841 gtttcgttga aaattgcagt aaagattgtg caagatgcat acaaagaaaa gatggccact 3901 gtttatcctg aaccccaaaa caaagaagaa tttgtctcct cccagatgta cagcactaat 3961 tatgaccaga tcctacctga ttgttatccg tggcctgcag aagtccagaa aatacagacc 4021 aaagtcaacc agggatccga ctacaaagac catgacggtg attataaaga tcatgacatc 4081 gactacaagg atgacgatga caagtagcgg ccgcgaagga tctgcgatcg ctccggtgcc 4141 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc 4201 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac 4261 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg 4321 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc 4381 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc 4441 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta 4501 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc 4561 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct 4621 gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagacgc caccatggag 4681 agcgacgaga gcggcctgcc cgccatggag atcgagtgcc gcatcaccgg caccctgaac 4741 ggcgtggagt tcgagctggt gggcggcgga gagggcaccc ccaagcaggg ccgcatgacc 4801 aacaagatga agagcaccaa aggcgccctg accttcagcc cctacctgct gagccacgtg 4861 atgggctacg gcttctacca cttcggcacc taccccagcg gctacgagaa ccccttcctg 4921 cacgccatca acaacggcgg ctacaccaac acccgcatcg agaagtacga ggacggcggc 4981 gtgctgcacg tgagcttcag ctaccgctac gaggccggcc gcgtgatcgg cgacttcaag 5041 gtggtgggca ccggcttccc cgaggacagc gtgatcttca ccgacaagat catccgcagc 5101 aacgccaccg tggagcacct gcaccccatg ggcgataacg tgctggtggg cagcttcgcc 5161 cgcaccttca gcctgcgcga cggcggctac tacagcttcg tggtggacag ccacatgcac 5221 ttcaagagcg ccatccaccc cagcatcctg cagaacgggg gccccatgtt cgccttccgc 5281 cgcgtggagg agctgcacag caacaccgag ctgggcatcg tggagtacca gcacgccttc 5341 aagaccccca tcgccttcgc cagatcccgc gctcagtcgt ccaattctgc cgtggacggc 5401 accgccggac ccggctccac cggatctcgc gagggcagag gaagtcttct aacatgcggt 5461 gacgtggagg agaatcccgg ccctatgacc gagtacaagc ccacggtgcg cctcgccacc 5521 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc 5581 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc 5641 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg 5701 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc 5761 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc 5821 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc 5881 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag 5941 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag 6001 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc 6061 atgacccgca agcccggtgc ctgaaatcaa cctctggatt acaaaatttg tgaaagattg 6121 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct 6181 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg 6241 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact 6301 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc 6361 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc 6421 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa 6481 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc 6541 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg 6601 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg 6661 gccgcctccc cgcctggtac ctttaagacc aatgacttac aaggcagctg tagatcttag 6721 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagataaga 6781 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc 6841 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt 6901 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc 6961 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact 7021 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta 7081 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 7141 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc 7201 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 7261 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag 7321 aagtagtgag gaggcttttt tggaggccta gacttttgca gagaccaaat tcgtaatcat 7381 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 7441 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 7501 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 7561 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 7621 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 7681 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 7741 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 7801 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 7861 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 7921 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 7981 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 8041 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 8101 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 8161 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 8221 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 8281 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 8341 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 8401 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 8461 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 8521 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 8581 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 8641 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 8701 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 8761 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 8821 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 8881 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 8941 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 9001 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 9061 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 9121 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 9181 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 9241 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 9301 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 9361 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 9421 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 9481 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta 9541 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg 9601 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt 9661 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg 9721 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt 9781 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg 9841 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 9901 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 9961 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg //