LOCUS V037583 9010 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V037583 VERSION V037583 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..3036 /label="TNFSF14(NM_003807)" /note="TNFSF14(NM_003807)" /gene="TNFSF14" CDS 3043..3108 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3152..3363 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3376..3644 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3673..3682 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3703..4365 /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 4435..4488 /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 4489..5088 /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 5089..5677 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5751..5984 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6056..6177 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6217..6352 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6385..6401) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6409..6425 /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(6433..6463) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6478..6499 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6787..7375) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7546..8406) /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(8407..8511) /gene="bla" /label="AmpR promoter" primer_bind 8985..9001 /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 aggagagtgt cgtacggccc 2341 tcagtgtttg tggtggatgg acagaccgac atcccattca cgaggctggg acgaagccac 2401 cggagacagt cgtgcagtgt ggcccgggtg ggtctgggtc tcttgctgtt gctgatgggg 2461 gccgggctgg ccgtccaagg ctggttcctc ctgcagctgc actggcgtct aggagagatg 2521 gtcacccgcc tgcctgacgg acctgcaggc tcctgggagc agctgataca agagcgaagg 2581 tctcacgagg tcaacccagc agcgcatctc acaggggcca actccagctt gaccggcagc 2641 ggggggccgc tgttatggga gactcagctg ggcctggcct tcctgagggg cctcagctac 2701 cacgatgggg cccttgtggt caccaaagct ggctactact acatctactc caaggtgcag 2761 ctgggcggtg tgggctgccc gctgggcctg gccagcacca tcacccacgg cctctacaag 2821 cgcacacccc gctaccccga ggagctggag ctgttggtca gccagcagtc accctgcgga 2881 cgggccacca gcagctcccg ggtctggtgg gacagcagct tcctgggtgg tgtggtacac 2941 ctggaggctg gggagaaggt ggtcgtccgt gtgctggatg aacgcctggt tcgactgcgt 3001 gatggtaccc ggtcttactt cggggctttc atggtgggat ccgactacaa agaccatgac 3061 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta gcggccgcga 3121 aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc 3181 gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta 3241 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg 3301 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca 3361 cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc 3421 cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact 3481 gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc 3541 ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa 3601 ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg 3661 cctacgctag acgccaccat ggagagcgac gagagcggcc tgcccgccat ggagatcgag 3721 tgccgcatca ccggcaccct gaacggcgtg gagttcgagc tggtgggcgg cggagagggc 3781 acccccaagc agggccgcat gaccaacaag atgaagagca ccaaaggcgc cctgaccttc 3841 agcccctacc tgctgagcca cgtgatgggc tacggcttct accacttcgg cacctacccc 3901 agcggctacg agaacccctt cctgcacgcc atcaacaacg gcggctacac caacacccgc 3961 atcgagaagt acgaggacgg cggcgtgctg cacgtgagct tcagctaccg ctacgaggcc 4021 ggccgcgtga tcggcgactt caaggtggtg ggcaccggct tccccgagga cagcgtgatc 4081 ttcaccgaca agatcatccg cagcaacgcc accgtggagc acctgcaccc catgggcgat 4141 aacgtgctgg tgggcagctt cgcccgcacc ttcagcctgc gcgacggcgg ctactacagc 4201 ttcgtggtgg acagccacat gcacttcaag agcgccatcc accccagcat cctgcagaac 4261 gggggcccca tgttcgcctt ccgccgcgtg gaggagctgc acagcaacac cgagctgggc 4321 atcgtggagt accagcacgc cttcaagacc cccatcgcct tcgccagatc ccgcgctcag 4381 tcgtccaatt ctgccgtgga cggcaccgcc ggacccggct ccaccggatc tcgcgagggc 4441 agaggaagtc ttctaacatg cggtgacgtg gaggagaatc ccggccctat gaccgagtac 4501 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc 4561 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag 4621 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg 4681 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg 4741 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc 4801 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc 4861 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg 4921 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg 4981 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg 5041 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaaa tcaacctctg 5101 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta 5161 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt 5221 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc 5281 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt 5341 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg 5401 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac 5461 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc 5521 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac 5581 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct 5641 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac 5701 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct 5761 aattcactcc caacgaagat aagatctgct ttttgcttgt actgggtctc tctggttaga 5821 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata 5881 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta 5941 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca 6001 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt 6061 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 6121 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 6181 tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt 6241 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc 6301 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt 6361 tgcagagacc aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct 6421 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 6481 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 6541 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 6601 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 6661 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 6721 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 6781 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 6841 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 6901 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 6961 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 7021 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 7081 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 7141 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 7201 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 7261 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 7321 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 7381 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 7441 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 7501 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 7561 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 7621 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 7681 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 7741 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 7801 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 7861 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 7921 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 7981 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 8041 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 8101 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 8161 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 8221 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 8281 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 8341 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 8401 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 8461 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 8521 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 8581 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 8641 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 8701 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 8761 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt 8821 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg 8881 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 8941 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 9001 tgccaagctg //