LOCUS V045429 9121 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045429 VERSION V045429 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" regulatory 2809..2818 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 2818..2847 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /transl_table=1 /translation="EQKLISEEDL" /label="Myc" CDS 2848..3792 /label="Ddrgk1(NM_029832)" /note="Ddrgk1(NM_029832)" /gene="Ddrgk1" misc_feature 3824..4410 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /label="IRES2" CDS 4430..5029 /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 5043..5631 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5838..6471 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6599..6615) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6623..6639 /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(6647..6677) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6692..6713 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7001..7589) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7760..8620) /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(8621..8725) /gene="bla" /label="AmpR promoter" polyA_signal 8773..8907 /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 tctatttccg gtgaattcgc caccatggag 2821 cagaagctga tctcagagga ggacctgatg gtggggccct gggtgtatct ggtggcggca 2881 gttttgctca tcggcctgat cctcttcctg actcgcagcc ggggtcgggc ggcagcagct 2941 gacggagaac cactgcacaa tgaggaagag agggcaggag caggccaggt aggccgctct 3001 ttgccccagg agtctgaaga acagagaact ggaagcagac cccggcgtcg gagggacttg 3061 ggcagccgtc tacaggccca gcgtcgagcc cagcgagtgg cctgggaaga cggggatgag 3121 aatgtgggtc aaactgttat tccagcccag gaggaagaag gcattgagaa gccagcagaa 3181 gttcacccaa cagggaaaat tggagccaag aaactacgga agctagagga aaaacaggct 3241 cgaaaggctc agcgagaggc agaggaggct gaacgtgaag aacggaaacg cctagagtcc 3301 caacgtgagg ccgaatggaa gaaggaagag gaacggcttc gcctgaagga agaacagaag 3361 gaggaggaag agaggaaggc tcaggaggag caggcccggc gggagcacga ggagtacctg 3421 aaactgaagg aggccttcgt ggtagaagaa gaaggtgtta gcgaaaccat gactgaggag 3481 cagtctcaca gcttcctgac agaattcatc aattacatca agaagtccaa ggttgtgctt 3541 ttggaagatc tggctttcca gatgggccta aggactcagg acgccataaa ccgcatccag 3601 gacctgctga cggaggggac tctaacaggt gtgattgacg accggggcaa gtttatctac 3661 ataaccccag aggaactggc tgccgtggcc aatttcatcc gacagcgggg ccgggtgtcc 3721 atcacagagc ttgcccaggc cagcaactcc ctcatctcct ggggccagga cctccctgcc 3781 caggcttcag cctgaactag ttctagagcg gccgcggatc ccgcccctct ccctcccccc 3841 cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt 3901 tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct 3961 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga 4021 atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga 4081 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac 4141 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag 4201 ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc 4261 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg 4321 tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt 4381 gaaaaacacg atgataagct tgccacaacc cacaaggaga cgaccttcca tgaccgagta 4441 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc 4501 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga 4561 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt 4621 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc 4681 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc 4741 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt 4801 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt 4861 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc 4921 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt 4981 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctaga cgcgtctgga 5041 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg 5101 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc 5161 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt 5221 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca 5281 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc 5341 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc 5401 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc 5461 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc 5521 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc 5581 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaatta 5641 attctgcagt cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac 5701 caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac 5761 acctcaggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt 5821 aaaagaaaag aggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga 5881 tctgtggatc taccacacac aaggctactt ccctgattag cagaactaca caccagggcc 5941 aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag ttgagccaga 6001 taaggtagaa gaggccaata aaggagagaa caccagcttg ttacaccctg tgagcctgca 6061 tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc gcctagcatt 6121 tcatcacgtg gcccgagagc tgcatccgga gtacttcaag aactgctgat atcgagcttg 6181 ctacaaggga ctttccgctg gggactttcc agggaggcgt ggcctgggcg ggactgggga 6241 gtggcgagcc ctcagatcct gcatataagc agctgctttt tgcctgtact gggtctctct 6301 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc 6361 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg 6421 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt 6481 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga 6541 ggccttgaca ttgctagcgt ttaccgtcga cctctagcta gagcttggcg taatcatggt 6601 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 6661 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 6721 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 6781 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 6841 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 6901 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 6961 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 7021 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 7081 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 7141 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 7201 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 7261 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 7321 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 7381 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 7441 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 7501 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 7561 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 7621 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 7681 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 7741 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 7801 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 7861 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 7921 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 7981 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 8041 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 8101 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 8161 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 8221 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 8281 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 8341 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 8401 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 8461 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 8521 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 8581 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 8641 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 8701 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg 8761 gatcgggaga tcaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 8821 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 8881 atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca aaatcatccc 8941 aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca tttactctaa 9001 acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact 9061 acaaacttag tagtttttaa agaaattgta tttgttaaat atgtactaca aacttagtag 9121 t //