LOCUS V035882 8524 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035882 VERSION V035882 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..3378 /label="ST8SIA4(NM_005668)" /note="ST8SIA4(NM_005668)" /gene="ST8SIA4" CDS 3385..3450 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3494..3705 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3718..3986 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4013..4612 /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 4619..5207 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5281..5514 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5586..5707 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5726..5861 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5899..5915) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5923..5939 /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(5947..5977) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5992..6013 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6301..6889) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7060..7920) /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(7921..8025) /gene="bla" /label="AmpR promoter" primer_bind 8499..8515 /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 catgcgctcc attaggaaga ggtggacgat ctgcacaata 2341 agtctgctcc tgatctttta taagacaaaa gaaatagcaa gaactgagga gcaccaggag 2401 acgcaactca tcggagatgg tgaattgtct ttgagtcggt cacttgtcaa tagctctgat 2461 aaaatcattc gaaaggctgg ctcttcaatc ttccagcaca atgtagaagg ttggaaaatc 2521 aattcctctt tggtcctaga gataaggaag aacatacttc gtttcttaga tgcagaacga 2581 gatgtgtcag tggtcaagag cagttttaag cctggtgatg tcatacacta tgtgcttgac 2641 aggcgccgga cactaaacat ttctcatgat ctacatagcc tcctacctga agtttcacca 2701 atgaagaatc gcaggtttaa gacctgtgca gttgttggaa attctggcat tctgttagac 2761 agtgaatgtg gaaaggagat tgacagtcac aattttgtaa taaggtgtaa tctagctcct 2821 gtggtggagt ttgctgcaga tgtgggaact aaatcagatt ttattaccat gaatccatca 2881 gttgtacaaa gagcatttgg aggctttcga aatgagagtg acagagaaaa atttgtgcat 2941 agactttcca tgctgaatga cagtgtcctt tggattcctg ctttcatggt caaaggagga 3001 gagaagcacg tggagtgggt taatgcatta atccttaaga ataaactgaa agtgcgaact 3061 gcctatccgt cattgagact tattcatgct gtcagaggtt actggctgac caacaaagtt 3121 cctatcaaaa gacccagcac aggtcttctc atgtatacac ttgccacaag attctgtgat 3181 gaaattcacc tgtatggatt ctggcccttc cctaaggatt taaatggaaa agcggtcaaa 3241 tatcattatt atgatgactt aaaatatagg tacttttcca atgcaagccc tcacagaatg 3301 ccattagaat tcaaaacatt aaatgtgcta cataatagag gagctctaaa actgacaaca 3361 ggaaagtgtg taaagcaagg atccgactac aaagaccatg acggtgatta taaagatcat 3421 gacatcgact acaaggatga cgatgacaag tagcggccgc gaaggatctg cgatcgctcc 3481 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg 3541 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc 3601 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc 3661 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg 3721 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg 3781 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta 3841 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga 3901 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg 3961 ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct agatgaccga 4021 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct 4081 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat 4141 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa 4201 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga 4261 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct 4321 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg 4381 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt 4441 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc 4501 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga 4561 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gagtcgacaa 4621 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc 4681 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat 4741 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg 4801 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg 4861 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat 4921 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt 4981 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc 5041 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa 5101 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg 5161 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa 5221 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac 5281 tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc 5341 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 5401 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 5461 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta 5521 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg 5581 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 5641 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 5701 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc 5761 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc 5821 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg 5881 cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 5941 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 6001 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 6061 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 6121 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 6181 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 6241 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 6301 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 6361 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 6421 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 6481 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 6541 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 6601 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 6661 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 6721 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 6781 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 6841 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 6901 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 6961 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 7021 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 7081 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 7141 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 7201 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 7261 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 7321 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 7381 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 7441 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 7501 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 7561 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 7621 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 7681 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 7741 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 7801 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 7861 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 7921 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 7981 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 8041 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 8101 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 8161 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 8221 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 8281 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 8341 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 8401 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 8461 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 8521 gctg //