LOCUS V032996 9541 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032996 VERSION V032996 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 1..634 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" misc_feature 681..806 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1303..1536 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1721..1765 /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 2028..2143 /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" /label="cPPT/CTS" enhancer 2201..2504 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 2505..2708 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2829..2855 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 2862..3704 /label="Mapre3(NM_133350)" /note="Mapre3(NM_133350)" /gene="Mapre3" CDS 3705..3758 /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 4257..4298 /codon_start=1 /product="epitope tag from simian virus 5" /transl_table=1 /translation="GKPIPNPLLGLDST" /label="V5 tag" promoter 4329..4828 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4849..5448 /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 5462..6050 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6257..6890 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7019..7035) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7043..7059 /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(7067..7097) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7112..7133 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7421..8009) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8180..9040) /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(9041..9145) /gene="bla" /label="AmpR promoter" polyA_signal 9193..9327 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag 2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa 2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg 2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt 2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg 2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc 2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc 2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca 2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta 2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa 2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc 2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag 2821 ccaccatgta cccatacgat gttccagatt acgctgaatt catggctgtc aatgtgtact 2881 ccacttctgt gaccagtgag aatctgagtc gccatgatat gcttgcatgg gtcaatgact 2941 ctctgcacct caattatacc aagattgaac agctctgttc aggggcagcc tactgccagt 3001 tcatggacat gctcttccct ggctgtgttc acttgaggaa ggtcaagttc caggccaaac 3061 tagaacacga gtacatccac aacttcaagg tgctgcaagc agctttcaag aagatgggtg 3121 ttgacaaaat cattcccgta gagaagttag tgaaaggaaa attccaagat aattttgagt 3181 ttatacagtg gtttaagaaa ttctttgacg caaactatga tggaaaggat tacaaccctc 3241 tgctggcgcg gcagggccag gacgtagcac cacctcctaa cccaggtgat cagatcttca 3301 acaaatccaa gaaactcatt ggcacagcag ttccgcagag gacgtccccc acaggcccca 3361 agaacatgca gacctctgga cgactcagca acgtggctcc gccctgcatc ctccggaaga 3421 atcccccctc agcccgaaac ggtggccatg aggctgacgc ccagatcctc gagcttaacc 3481 agcagctgct ggacttgaag ctgaccgtag acgggcttga gaaagaacga gatttctatt 3541 tcagcaaatt gcgagacatc gagctgatct gccaggaaca tgagagcgag aacagccccg 3601 tcatctcggg catcattggc attctctatg ccacggagga gggatttgca ccccctgagg 3661 atgatgagat tgaagaacac caacaggaag accaggacga gtacgagggc agaggaagtc 3721 ttctaacatg cggtgacgtg gaggagaatc ccggccctat ggcctctggt gtggctgtct 3781 ctgatggtgt catcaaggtg ttcaatgaca tgaaggttcg caagtcttca acaccagaag 3841 aagtgaagaa acgcaagaag gcggtgctct tttgcctgag tgaggacaag aagaacatca 3901 tcctggagga gggcaaggag atcctggtag gagatgtggg gcagactgtg gacgacccct 3961 acaccacttt tgtcaagatg ctgccagaca aggactgccg ctatgcactc tatgatgcaa 4021 cctatgagac caaggagagc aagaaggagg acctggtgtt catcttctgg gcccccgaga 4081 atgcacccct caagagcaaa atgatctatg ccagctccaa ggatgccatc aagaagaagc 4141 tgacaggaat caagcatgaa ttacaagcta actgctacga ggaggtcaag gaccgctgca 4201 ccctggcaga gaaactaggt ggcagcgccg tcatttccct ggagggcaag cctttgggta 4261 agcctatccc taaccctctc ctcggtctcg attctacgta ggatcccgcg actctagata 4321 attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg ctttagcagc 4381 cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt ccacatccac 4441 cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt ctactcctcc 4501 cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga 4561 agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt 4621 aggcctttgg ggcagcggcc aatagcagct ttgctccttc gctttctggg ctcagaggct 4681 gggaaggggt gggtccgggg gcgggctcag gggcgggctc aggggcgggg cgggcgcccg 4741 aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt 4801 ctcctcttcc tcatctccgg gcctttcgac ctgcagccca agcttaccat gaccgagtac 4861 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc 4921 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag 4981 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg 5041 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg 5101 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc 5161 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc 5221 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg 5281 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg 5341 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg 5401 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacc gcgtctggaa 5461 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 5521 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 5581 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 5641 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 5701 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 5761 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 5821 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct 5881 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 5941 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 6001 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattaa 6061 ttctgcagtc gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc 6121 aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca 6181 cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta 6241 aaagaaaaga ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat 6301 ctgtggatct accacacaca aggctacttc cctgattagc agaactacac accagggcca 6361 ggggtcagat atccactgac ctttggatgg tgctacaagc tagtaccagt tgagccagat 6421 aaggtagaag aggccaataa aggagagaac accagcttgt tacaccctgt gagcctgcat 6481 gggatggatg acccggagag agaagtgtta gagtggaggt ttgacagccg cctagcattt 6541 catcacgtgg cccgagagct gcatccggag tacttcaaga actgctgata tcgagcttgc 6601 tacaagggac tttccgctgg ggactttcca gggaggcgtg gcctgggcgg gactggggag 6661 tggcgagccc tcagatcctg catataagca gctgcttttt gcctgtactg ggtctctctg 6721 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc 6781 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg 6841 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc 6901 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag 6961 gccttgacat tgctagcgtt ttaccgtcga cctctagcta gagcttggcg taatcatggt 7021 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 7081 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 7141 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 7201 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 7261 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 7321 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 7381 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 7441 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 7501 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 7561 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 7621 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 7681 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 7741 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 7801 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 7861 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 7921 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 7981 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 8041 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 8101 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 8161 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 8221 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 8281 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 8341 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 8401 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 8461 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 8521 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 8581 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 8641 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 8701 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 8761 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 8821 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 8881 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 8941 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 9001 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 9061 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 9121 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg 9181 gatcgggaga tcaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 9241 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 9301 atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca aaatcatccc 9361 aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca tttactctaa 9421 acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact 9481 acaaacttag tagtttttaa agaaattgta tttgttaaat atgtactaca aacttagtag 9541 t //