LOCUS V020423 9646 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020423 VERSION V020423 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 2820..4313 /label="TRIM22(NM_006074)" /note="TRIM22(NM_006074)" /gene="TRIM22" CDS 4314..4337 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" misc_feature 4349..4935 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /label="IRES2" CDS 4955..5554 /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 5568..6156 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6363..6996 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7124..7140) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7148..7164 /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(7172..7202) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7217..7238 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7526..8114) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8285..9145) /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(9146..9250) /gene="bla" /label="AmpR promoter" polyA_signal 9298..9432 /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 gtgaattcct cgagccacca 2821 tggatttctc agtaaaggta gacatagaga aggaggtgac ctgccccatc tgcctggagc 2881 tcctgacaga acctctgagc ctagattgtg gccacagctt ctgccaagcc tgcatcactg 2941 caaagatcaa ggagtcagtg atcatctcaa gaggggaaag cagctgtcct gtgtgtcaga 3001 ccagattcca gcctgggaac ctccgaccta atcggcatct ggccaacata gttgagagag 3061 tcaaagaggt caagatgagc ccacaggagg ggcagaagag agatgtctgt gagcaccatg 3121 gaaaaaaact ccagatcttc tgtaaggagg atggaaaagt catttgctgg gtttgtgaac 3181 tgtctcagga acaccaaggt caccaaacat tccgcataaa cgaggtggtc aaggaatgtc 3241 aggaaaagct gcaggtagcc ctgcagaggc tgataaagga ggatcaagag gctgagaagc 3301 tggaagatga catcagacaa gagagaaccg cctggaagaa ttatatccag atcgagagac 3361 agaagattct gaaagggttc aatgaaatga gagtcatctt ggacaatgag gagcagagag 3421 agctgcaaaa gctggaggaa ggtgaggtga atgtgctgga taacctggca gcagctacag 3481 accagctggt ccagcagagg caggatgcca gcacgctcat ctcagatctc cagcggaggt 3541 tgaggggatc gtcagtagag atgctgcagg atgtgattga cgtcatgaaa aggagtgaaa 3601 gctggacatt gaagaagcca aaatctgttt ccaagaaact aaagagtgta ttccgagtac 3661 cagatctgag tgggatgctg caagttctta aagagctgac agatgtccag tactactggg 3721 tggacgtgat gctgaatcca ggcagtgcca cttcgaatgt tgctatttct gtggatcaga 3781 gacaagtgaa aactgtacgc acctgcacat ttaagaattc aaatccatgt gatttttctg 3841 cttttggtgt cttcggctgc caatatttct cttcggggaa atattactgg gaagtagatg 3901 tgtctggaaa gattgcctgg atcctgggcg tacacagtaa aataagtagt ctgaataaaa 3961 ggaagagctc tgggtttgct tttgatccaa gtgtaaatta ttcaaaagtt tactccagat 4021 atagacctca atatggctac tgggttatag gattacagaa tacatgtgaa tataatgctt 4081 ttgaggactc ctcctcttct gatcccaagg ttttgactct ctttatggct gtgcctccct 4141 gtcgtattgg ggttttccta gactatgagg caggcattgt ctcatttttc aatgtcacaa 4201 accacggagc actcatctac aagttctctg gatgtcgctt ttctcgacct gcttatccgt 4261 atttcaatcc ttggaactgc ctagtcccca tgactgtgtg cccaccgagc tccgattaca 4321 aggacgacga tgacaagtga ggatcccgcc cctctccctc ccccccccct aacgttactg 4381 gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat 4441 tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc 4501 ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag 4561 cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc 4621 ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac 4681 ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca 4741 aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt 4801 gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa 4861 aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat 4921 aagcttgcca caacccacaa ggagacgacc ttccatgacc gagtacaagc ccacggtgcg 4981 cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga 5041 ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct 5101 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga 5161 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc 5221 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat 5281 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg 5341 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga 5401 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc 5461 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg 5521 cacctggtgc atgacccgca agcccggtgc ctagacgcgt ctggaacaat caacctctgg 5581 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat 5641 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt 5701 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca 5761 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg 5821 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg 5881 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca 5941 attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca 6001 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc 6061 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc 6121 agacgagtcg gatctccctt tgggccgcct ccccgcctgg aattaattct gcagtcgaga 6181 cctagaaaaa catggagcaa tcacaagtag caatacagca gctaccaatg ctgattgtgc 6241 ctggctagaa gcacaagagg aggaggaggt gggttttcca gtcacacctc aggtaccttt 6301 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagagggg 6361 actggaaggg ctaattcact cccaacgaag acaagatatc cttgatctgt ggatctacca 6421 cacacaaggc tacttccctg attagcagaa ctacacacca gggccagggg tcagatatcc 6481 actgaccttt ggatggtgct acaagctagt accagttgag ccagataagg tagaagaggc 6541 caataaagga gagaacacca gcttgttaca ccctgtgagc ctgcatggga tggatgaccc 6601 ggagagagaa gtgttagagt ggaggtttga cagccgccta gcatttcatc acgtggcccg 6661 agagctgcat ccggagtact tcaagaactg ctgatatcga gcttgctaca agggactttc 6721 cgctggggac tttccaggga ggcgtggcct gggcgggact ggggagtggc gagccctcag 6781 atcctgcata taagcagctg ctttttgcct gtactgggtc tctctggtta gaccagatct 6841 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc 6901 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc 6961 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta 7021 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggcct tgacattgct 7081 agcgtttacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg 7141 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta 7201 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg 7261 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga 7321 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 7381 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 7441 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 7501 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 7561 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 7621 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 7681 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 7741 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 7801 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 7861 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 7921 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 7981 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 8041 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 8101 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 8161 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 8221 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 8281 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 8341 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 8401 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 8461 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 8521 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 8581 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 8641 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 8701 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 8761 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 8821 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 8881 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 8941 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 9001 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 9061 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 9121 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 9181 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 9241 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatcaac 9301 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat 9361 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat 9421 catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact tcccacccca 9481 taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg tgattcctct 9541 gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa cttagtagtt 9601 tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagt //