LOCUS V022945 9230 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022945 VERSION V022945 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 2854..3906 /label="THOC3(NM_032361)" /note="THOC3(NM_032361)" /gene="THOC3" misc_feature 3933..4519 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /label="IRES2" CDS 4539..5138 /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 5152..5740 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5947..6580 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6708..6724) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6732..6748 /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(6756..6786) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6801..6822 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7110..7698) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7869..8729) /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(8730..8834) /gene="bla" /label="AmpR promoter" polyA_signal 8882..9016 /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 caccatggaa 2821 caaaaactca tctcagaaga ggatctgctc gagatggcgg tccccgctgc agccatgggg 2881 ccctcggcgt tgggccagag cggccccggc tcgatggccc cgtggtgctc agtgagcagc 2941 ggcccgtcgc gctacgtgct tgggatgcag gagctgttcc ggggccacag caagacgcgc 3001 gagttcctgg cgcacagcgc caaggtgcac tcggtggcct ggagttgcga cgggcgtcgc 3061 ctagcctcgg ggtccttcga caagacggcc agcgtcttct tgctggagaa ggaccggttg 3121 gtcaaagaaa acaattatcg gggacatggg gatagtgtgg accagctttg ttggcatcca 3181 agtaatcctg acctatttgt tacggcgtcc ggagataaaa ccattcgcat ctgggatgtg 3241 aggactacaa aatgcattgc cactgtgaac actaaagggg agaacattaa tatctgctgg 3301 agtcctgatg ggcagaccat tgctgtaggc aacaaggatg atgtggtgac ctttattgat 3361 gccaagacac accgttccaa agcagaagag cagttcaagt tcgaggtcaa cgaaatctcc 3421 tggaacaatg acaataatat gttcttcctg acaaatggca atggttgtat caacatcctc 3481 agctacccag aactgaagcc tgtgcagtcc atcaacgccc atccttccaa ctgcatctgt 3541 atcaagtttg accccatggg gaagtacttt gccacaggaa gtgcagatgc tttggtcagc 3601 ctctgggatg tggatgagtt agtgtgtgtt cggtgctttt ccaggctgga ttggcctgta 3661 agaaccctca gtttcagcca tgatgggaaa atgctggcgt cagcatcgga agatcatttt 3721 attgacattg ctgaagtgga gacaggggac aaactatggg aggtacagtg tgagtctccg 3781 accttcacag tggcgtggca ccccaaaagg cctctgctgg catttgcctg tgatgacaaa 3841 gacggcaaat atgacagcag ccgggaagcc ggaactgtga agctgtttgg gcttcctaat 3901 gattcttagt tctagagcgg ccgcggatcc cgcccctctc cctccccccc ccctaacgtt 3961 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc 4021 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc 4081 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag 4141 gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg 4201 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat 4261 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga 4321 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc 4381 cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg 4441 ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga 4501 tgataagctt gccacaaccc acaaggagac gaccttccat gaccgagtac aagcccacgg 4561 tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg caccctcgcc gccgcgttcg 4621 ccgactaccc cgccacgcgc cacaccgtcg acccggaccg ccacatcgag cgggtcaccg 4681 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg 4741 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt 4801 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac 4861 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg 4921 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag 4981 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc 5041 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac 5101 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctagac gcgtctggaa caatcaacct 5161 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 5221 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 5281 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 5341 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 5401 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 5461 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 5521 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt 5581 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 5641 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 5701 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattaa ttctgcagtc 5761 gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc aatgctgatt 5821 gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca cctcaggtac 5881 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaaga 5941 ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat ctgtggatct 6001 accacacaca aggctacttc cctgattagc agaactacac accagggcca ggggtcagat 6061 atccactgac ctttggatgg tgctacaagc tagtaccagt tgagccagat aaggtagaag 6121 aggccaataa aggagagaac accagcttgt tacaccctgt gagcctgcat gggatggatg 6181 acccggagag agaagtgtta gagtggaggt ttgacagccg cctagcattt catcacgtgg 6241 cccgagagct gcatccggag tacttcaaga actgctgata tcgagcttgc tacaagggac 6301 tttccgctgg ggactttcca gggaggcgtg gcctgggcgg gactggggag tggcgagccc 6361 tcagatcctg catataagca gctgcttttt gcctgtactg ggtctctctg gttagaccag 6421 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc 6481 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga 6541 tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt 6601 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gccttgacat 6661 tgctagcgtt taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt 6721 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag 6781 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg 6841 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg 6901 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc 6961 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc 7021 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg 7081 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 7141 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 7201 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 7261 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 7321 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 7381 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 7441 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 7501 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt 7561 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 7621 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 7681 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 7741 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 7801 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 7861 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 7921 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 7981 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 8041 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 8101 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 8161 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 8221 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 8281 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 8341 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 8401 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 8461 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 8521 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 8581 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 8641 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 8701 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 8761 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 8821 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat 8881 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 8941 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 9001 ttatcatgtc tggatcaact ggataactca agctaaccaa aatcatccca aacttcccac 9061 cccataccct attaccactg ccaattacct gtggtttcat ttactctaaa cctgtgattc 9121 ctctgaatta ttttcatttt aaagaaattg tatttgttaa atatgtacta caaacttagt 9181 agtttttaaa gaaattgtat ttgttaaata tgtactacaa acttagtagt //