LOCUS V043720 9484 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V043720 VERSION V043720 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..4338 /label="HSPA9(NM_004134)" /note="HSPA9(NM_004134)" /gene="HSPA9" CDS 4345..4410 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4454..4665 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4678..4946 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4973..5572 /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 5579..6167 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6241..6474 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6546..6667 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6686..6821 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6859..6875) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6883..6899 /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(6907..6937) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6952..6973 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7261..7849) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8020..8880) /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(8881..8985) /gene="bla" /label="AmpR promoter" primer_bind 9459..9475 /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 catgataagt gccagccgag ctgcagcagc ccgtctcgtg 2341 ggcgccgcag cctcccgggg ccctacggcc gcccgccacc aggatagctg gaatggcctt 2401 agtcatgagg cttttagact tgtttcaagg cgggattatg catcagaagc aatcaaggga 2461 gcagttgttg gtattgattt gggtactacc aactcctgcg tggcagttat ggaaggtaaa 2521 caagcaaagg tgctggagaa tgccgaaggt gccagaacca ccccttcagt tgtggccttt 2581 acagcagatg gtgagcgact tgttggaatg ccggccaagc gacaggctgt caccaaccca 2641 aacaatacat tttatgctac caagcgtctc attggccggc gatatgatga tcctgaagta 2701 cagaaagaca ttaaaaatgt tccctttaaa attgtccgtg cctccaatgg tgatgcctgg 2761 gttgaggctc atgggaaatt gtattctccg agtcagattg gagcatttgt gttgatgaag 2821 atgaaagaga ctgcagaaaa ttacttgggg cacacagcaa aaaatgctgt gatcacagtc 2881 ccagcttatt tcaatgactc gcagagacag gccactaaag atgctggcca gatatctgga 2941 ctgaatgtgc ttcgggtgat taatgagccc acagctgctg ctcttgccta tggtctagac 3001 aaatcagaag acaaagtcat tgctgtatat gatttaggtg gtggaacttt tgatatttct 3061 atcctggaaa ttcagaaagg agtatttgag gtgaaatcca caaatgggga taccttctta 3121 ggtggggaag actttgacca ggccttgcta cggcacattg tgaaggagtt caagagagag 3181 acaggggttg atttgactaa agacaacatg gcacttcaga gggtacggga agctgctgaa 3241 aaggctaaat gtgaactctc ctcatctgtg cagactgaca tcaatttgcc ctatcttaca 3301 atggattctt ctggacccaa gcatttgaat atgaagttga cccgtgctca atttgaaggg 3361 attgtcactg atctaatcag aaggactatc gctccatgcc aaaaagctat gcaagatgca 3421 gaagtcagca agagtgacat aggagaagtg attcttgtgg gtggcatgac taggatgccc 3481 aaggttcagc agactgtaca ggatcttttt ggcagagccc caagtaaagc tgtcaatcct 3541 gatgaggctg tggccattgg agctgccatt cagggaggtg tgttggccgg cgatgtcacg 3601 gatgtgctgc tccttgatgt cactcccctg tctctgggta ttgaaactct aggaggtgtc 3661 tttaccaaac ttattaatag gaataccact attccaacca agaagagcca ggtattctct 3721 actgccgctg atggtcaaac gcaagtggaa attaaagtgt gtcagggtga aagagagatg 3781 gctggagaca acaaactcct tggacagttt actttgattg gaattccacc agcccctcgt 3841 ggagttcctc agattgaagt tacatttgac attgatgcca atgggatagt acatgtttct 3901 gctaaagata aaggcacagg acgtgagcag cagattgtaa tccagtcttc tggtggatta 3961 agcaaagatg atattgaaaa tatggttaaa aatgcagaga aatatgctga agaagaccgg 4021 cgaaagaagg aacgagttga agcagttaat atggctgaag gaatcattca cgacacagaa 4081 accaagatgg aagaattcaa ggaccaatta cctgctgatg agtgcaacaa gctgaaagaa 4141 gagatttcca aaatgaggga gctcctggct agaaaagaca gcgaaacagg agaaaatatt 4201 agacaggcag catcctctct tcagcaggca tcactgaagc tgttcgaaat ggcatacaaa 4261 aagatggcat ctgagcgaga aggctctgga agttctggca ctggggaaca aaaggaagat 4321 caaaaggagg aaaaacaggg atccgactac aaagaccatg acggtgatta taaagatcat 4381 gacatcgact acaaggatga cgatgacaag tagcggccgc gaaggatctg cgatcgctcc 4441 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg 4501 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc 4561 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc 4621 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg 4681 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg 4741 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta 4801 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga 4861 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg 4921 ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct agatgaccga 4981 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct 5041 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat 5101 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa 5161 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga 5221 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct 5281 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg 5341 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt 5401 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc 5461 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga 5521 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gagtcgacaa 5581 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc 5641 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat 5701 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg 5761 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg 5821 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat 5881 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt 5941 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc 6001 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa 6061 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg 6121 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa 6181 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac 6241 tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc 6301 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 6361 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 6421 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta 6481 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg 6541 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 6601 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 6661 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc 6721 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc 6781 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg 6841 cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 6901 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 6961 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 7021 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 7081 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 7141 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 7201 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 7261 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 7321 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 7381 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 7441 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 7501 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 7561 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 7621 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 7681 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 7741 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 7801 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 7861 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 7921 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 7981 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 8041 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 8101 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 8161 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 8221 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 8281 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 8341 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 8401 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 8461 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 8521 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 8581 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 8641 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 8701 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 8761 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 8821 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 8881 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 8941 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 9001 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 9061 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 9121 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 9181 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 9241 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 9301 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 9361 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 9421 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 9481 gctg //