LOCUS V022891 10042 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022891 VERSION V022891 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 2843..4717 /label="RMI1(NM_001358293)" /note="RMI1(NM_001358293)" /gene="RMI1" CDS 4724..4750 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 4754..4780 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 4784..4810 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" promoter 4830..5329 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5350..5949 /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 5963..6551 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6758..7391 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7520..7536) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7544..7560 /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(7568..7598) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7613..7634 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7922..8510) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8681..9541) /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(9542..9646) /gene="bla" /label="AmpR promoter" polyA_signal 9694..9828 /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 ctcaagcttc gaattcgcca ccatgaatgt gactagtatt gcattaagag ctgaaacttg 2881 gcttttagct gcatggcatg ttaaagtacc tccgatgtgg ctggaagctt gtattaactg 2941 gattcaagaa gaaaataata atgttaactt gagtcaggcc caaatgaata aacaagtgtt 3001 tgagcagtgg ctccttactg atctgaggga tttggagcat cctcttttac ccgatggcat 3061 tttagaaatt ccaaaaggag aattaaatgg attttatgct ctgcagatta attccttggt 3121 tgatgtaagt cagcctgcat actcccagat acagaagttg agaggaaaga atacaacaaa 3181 tgatctagtt acagctgaag cacaagtaac cccaaaacct tgggaagcaa agccttcacg 3241 aatgttgatg ctgcagctaa ctgatggaat cgtacaaata cagggaatgg aatatcagcc 3301 tattccaatt cttcatagtg atcttcctcc aggtacaaaa attttgattt atggaaatat 3361 atctttccgt cttggtgttc tcttattgaa accagaaaac gtgaaagtgt taggaggtga 3421 agtagatgct cttttagaag aatatgccca agaaaaagta cttgcaagat taatagggga 3481 acctgatctt gtagtttcag tcataccaaa caattctaac gaaaacattc ccagagttac 3541 agatgttcta gatcctgcat taggtccttc tgatgaagaa ctcttggcaa gtcttgatga 3601 aaatgatgag cttacagcaa ataatgacac ttcctcagaa cgatgtttca ccacaggtag 3661 ttcctcaaat accattccca caagacagtc aagttttgag ccagaatttg ttatttctcc 3721 aagaccaaaa gaggaaccat caaacctatc tatacatgta atggatggag aattagatga 3781 cttttcactg gaggaggcct tgcttttaga agaaactgtc cagaaagaac agatggaaac 3841 taaggaattg caaccattga cttttaacag aaatgccgat cgaagtatag agagattttc 3901 acataatcct aatactacga ataacttttc tttgacttgc aaaaatggaa acaataattg 3961 gagtgaaaaa aatgtatctg aacaaatgac taatgaagac aaatcatttg gttgtccatc 4021 tgttagagac caaaacagga gtattttttc agttcattgt aatgtaccct tagcccatga 4081 ttttacaaat aaagaaaaga acttagagac agataataaa ataaaacaaa ccagcagttc 4141 agatagccat tccttaaata ataaaatatt aaatagagag gtggtcaact atgtacagaa 4201 aaggaattca caaatttcta atgaaaatga ttgtaattta cagagttgtt ctttaagatc 4261 atcagagaat agcattaatc tttctattgc catggatttg tattctccac cctttgtcta 4321 tttgtctgtt ctaatggcca gcaaaccaaa ggaagttaca acagtgaaag tgaaagcatt 4381 tattgtaacc ttaactggaa atctctcaag ttctggtggt atttggagta taactgcaaa 4441 ggtgtctgat ggtactgcat atctagatgt agactttgtg gatgaaatac ttactagctt 4501 gatagggttc tcagtaccag aaatgaaaca gtcaaaaaag gatcctcttc aataccaaaa 4561 gttcctggaa gggttgcaga aatgtcaaag agatctaata gatttgtgct gtctaatgac 4621 tatttcattt aatccttcct tgtctaaagc aatggtactg gcattacaag atgttaatat 4681 ggaacacctt gagaatctaa agaagcggtt aaataaagga tcctacccat acgacgtacc 4741 tgattatgca acttatcctt atgatgtacc tgattatgct agctatcctt atgatgtccc 4801 agactacgca tagtctagat aattctaccg ggtaggggag gcgcttttcc caaggcagtc 4861 tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg gcctctggcc 4921 tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt 4981 cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc 5041 gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac 5101 cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt 5161 cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct 5221 caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa 5281 gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga cctgcagccc 5341 aagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc 5401 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc 5461 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc 5521 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc 5581 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag 5641 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg 5701 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag 5761 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc 5821 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc 5881 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc 5941 ggtgcctgac cgcgtctgga acaatcaacc tctggattac aaaatttgtg aaagattgac 6001 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt 6061 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt 6121 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt 6181 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg 6241 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg 6301 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct 6361 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt 6421 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc 6481 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc 6541 cgcctccccg cctggaatta attctgcagt cgagacctag aaaaacatgg agcaatcaca 6601 agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca agaggaggag 6661 gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct 6721 gtagatctta gccacttttt aaaagaaaag aggggactgg aagggctaat tcactcccaa 6781 cgaagacaag atatccttga tctgtggatc taccacacac aaggctactt ccctgattag 6841 cagaactaca caccagggcc aggggtcaga tatccactga cctttggatg gtgctacaag 6901 ctagtaccag ttgagccaga taaggtagaa gaggccaata aaggagagaa caccagcttg 6961 ttacaccctg tgagcctgca tgggatggat gacccggaga gagaagtgtt agagtggagg 7021 tttgacagcc gcctagcatt tcatcacgtg gcccgagagc tgcatccgga gtacttcaag 7081 aactgctgat atcgagcttg ctacaaggga ctttccgctg gggactttcc agggaggcgt 7141 ggcctgggcg ggactgggga gtggcgagcc ctcagatcct gcatataagc agctgctttt 7201 tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact 7261 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc 7321 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa 7381 aatctctagc agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa 7441 atgaatatca gagagtgaga ggccttgaca ttgctagcgt tttaccgtcg acctctagct 7501 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 7561 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 7621 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 7681 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 7741 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 7801 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 7861 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 7921 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 7981 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 8041 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 8101 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 8161 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 8221 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 8281 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 8341 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 8401 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 8461 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 8521 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 8581 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 8641 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 8701 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 8761 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 8821 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 8881 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 8941 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 9001 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 9061 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 9121 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 9181 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 9241 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 9301 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 9361 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 9421 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 9481 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 9541 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 9601 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 9661 aagtgccacc tgacgtcgac ggatcgggag atcaacttgt ttattgcagc ttataatggt 9721 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct 9781 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcaa ctggataact 9841 caagctaacc aaaatcatcc caaacttccc accccatacc ctattaccac tgccaattac 9901 ctgtggtttc atttactcta aacctgtgat tcctctgaat tattttcatt ttaaagaaat 9961 tgtatttgtt aaatatgtac tacaaactta gtagttttta aagaaattgt atttgttaaa 10021 tatgtactac aaacttagta gt //