LOCUS V046369 9004 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V046369 VERSION V046369 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..1920 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2010..2213 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2302..3858 /label="LAP3(NM_015907)" /note="LAP3(NM_015907)" /gene="LAP3" CDS 3865..3930 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3974..4185 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4198..4466 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4493..5092 /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 5099..5687 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5761..5994 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6066..6187 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6206..6341 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6379..6395) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6403..6419 /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(6427..6457) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6472..6493 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6781..7369) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7540..8400) /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(8401..8505) /gene="bla" /label="AmpR promoter" primer_bind 8979..8995 /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 attacaaaat tcaaaatttt 1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag 2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct 2281 agagctagcg aattcgccac catgttcttg ctgcctcttc cggctgcggg gcgagtagtc 2341 gtccgacgtc tggccgtgag acgtttcggg agccggagtc tctccaccgc agacatgacg 2401 aagggccttg ttttaggaat ctattccaaa gaaaaagaag atgatgtgcc acagttcaca 2461 agtgcaggag agaattttga taaattgtta gctggaaagc tgagagagac tttgaacata 2521 tctggaccac ctctgaaggc agggaagact cgaacctttt atggtctgca tcaggacttc 2581 cccagcgtgg tgctagttgg cctcggcaaa aaggcagctg gaatcgacga acaggaaaac 2641 tggcatgaag gcaaagaaaa catcagagct gctgttgcag cggggtgcag gcagattcaa 2701 gacctggagc tctcgtctgt ggaggtggat ccctgtggag acgctcaggc tgctgcggag 2761 ggagcggtgc ttggtctcta tgaatacgat gacctaaagc aaaaaaagaa gatggctgtg 2821 tcggcaaagc tctatggaag tggggatcag gaggcctggc agaaaggagt cctgtttgct 2881 tctgggcaga acttggcacg ccaattgatg gagacgccag ccaatgagat gacgccaacc 2941 agatttgctg aaattattga gaagaatctc aaaagtgcta gtagtaaaac cgaggtccat 3001 atcagaccca agtcttggat tgaggaacag gcaatgggat cattcctcag tgtggccaaa 3061 ggatctgacg agcccccagt cttcttggaa attcactaca aaggcagccc caatgcaaac 3121 gaaccacccc tggtgtttgt tgggaaagga attacctttg acagtggtgg tatctccatc 3181 aaggcttctg caaatatgga cctcatgagg gctgacatgg gaggagctgc aactatatgc 3241 tcagccatcg tgtctgctgc aaagcttaat ttgcccatta atattatagg tctggcccct 3301 ctttgtgaaa atatgcccag cggcaaggcc aacaagccgg gggatgttgt tagagccaaa 3361 aacgggaaga ccatccaggt tgataacact gatgctgagg ggaggctcat actggctgat 3421 gcgctctgtt acgcacacac gtttaacccg aaggtcatcc tcaatgccgc caccttaaca 3481 ggtgccatgg atgtagcttt gggatcaggt gccactgggg tctttaccaa ttcatcctgg 3541 ctctggaaca aactcttcga ggccagcatt gaaacagggg accgtgtctg gaggatgcct 3601 ctcttcgaac attatacaag acaggttgta gattgccagc ttgctgatgt taacaacatt 3661 ggaaaataca gatctgcagg agcatgtaca gctgcagcat tcctgaaaga attcgtaact 3721 catcctaagt gggcacattt agacatagca ggcgtgatga ccaacaaaga tgaagttccc 3781 tatctacgga aaggcatgac tgggaggccc acaaggactc tcattgagtt cttacttcgt 3841 ttcagtcaag acaatgctgg atccgactac aaagaccatg acggtgatta taaagatcat 3901 gacatcgact acaaggatga cgatgacaag tagcggccgc gaaggatctg cgatcgctcc 3961 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg 4021 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc 4081 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc 4141 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg 4201 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg 4261 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta 4321 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga 4381 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg 4441 ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct agatgaccga 4501 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct 4561 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat 4621 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa 4681 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga 4741 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct 4801 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg 4861 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt 4921 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc 4981 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga 5041 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gagtcgacaa 5101 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc 5161 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat 5221 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg 5281 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg 5341 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat 5401 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt 5461 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc 5521 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa 5581 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg 5641 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa 5701 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac 5761 tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc 5821 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 5881 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 5941 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta 6001 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg 6061 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 6121 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 6181 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc 6241 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc 6301 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg 6361 cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 6421 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 6481 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 6541 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 6601 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 6661 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 6721 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 6781 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 6841 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 6901 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 6961 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 7021 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 7081 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 7141 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 7201 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 7261 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 7321 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 7381 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 7441 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 7501 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 7561 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 7621 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 7681 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 7741 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 7801 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 7861 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 7921 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 7981 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 8041 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 8101 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 8161 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 8221 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 8281 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 8341 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 8401 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 8461 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 8521 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 8581 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 8641 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 8701 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 8761 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 8821 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 8881 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 8941 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 9001 gctg //