LOCUS V021149 9822 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V021149 VERSION V021149 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 2820..2829 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 2829..2894 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 2895..4547 /label="CBS(NM_000071)" /note="CBS(NM_000071)" /gene="CBS" promoter 4610..5109 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5130..5729 /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 5743..6331 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6538..7171 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7300..7316) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7324..7340 /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(7348..7378) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7393..7414 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7702..8290) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8461..9321) /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(9322..9426) /gene="bla" /label="AmpR promoter" polyA_signal 9474..9608 /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 ccaccatgga ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg 2881 atgacgatga caagatgcct tctgagaccc cccaggcaga agtggggccc acaggctgcc 2941 cccaccgctc agggccacac tcggcgaagg ggagcctgga gaaggggtcc ccagaggata 3001 aggaagccaa ggagcccctg tggatccggc ccgatgctcc gagcaggtgc acctggcagc 3061 tgggccggcc tgcctccgag tccccacatc accacactgc cccggcaaaa tctccaaaaa 3121 tcttgccaga tattctgaag aaaatcgggg acacccctat ggtcagaatc aacaagattg 3181 ggaagaagtt cggcctgaag tgtgagctct tggccaagtg tgagttcttc aacgcgggcg 3241 ggagcgtgaa ggaccgcatc agcctgcgga tgattgagga tgctgagcgc gacgggacgc 3301 tgaagcccgg ggacacgatt atcgagccga catccgggaa caccgggatc gggctggccc 3361 tggctgcggc agtgaggggc tatcgctgca tcatcgtgat gccagagaag atgagctccg 3421 agaaggtgga cgtgctgcgg gcactggggg ctgagattgt gaggacgccc accaatgcca 3481 ggttcgactc cccggagtca cacgtggggg tggcctggcg gctgaagaac gaaatcccca 3541 attctcacat cctagaccag taccgcaacg ccagcaaccc cctggctcac tacgacacca 3601 ccgctgatga gatcctgcag cagtgtgatg ggaagctgga catgctggtg gcttcagtgg 3661 gcacgggcgg caccatcacg ggcattgcca ggaagctgaa ggagaagtgt cctggatgca 3721 ggatcattgg ggtggatccc gaagggtcca tcctcgcaga gccggaggag ctgaaccaga 3781 cggagcagac aacctacgag gtggaaggga tcggctacga cttcatcccc acggtgctgg 3841 acaggacggt ggtggacaag tggttcaaga gcaacgatga ggaggcgttc acctttgccc 3901 gcatgctgat cgcgcaagag gggctgctgt gcggtggcag tgctggcagc acggtggcgg 3961 tggccgtgaa ggctgcgcag gagctgcagg agggccagcg ctgcgtggtc attctgcccg 4021 actcagtgcg gaactacatg accaagttcc tgagcgacag gtggatgctg cagaagggct 4081 ttctgaagga ggaggacctc acggagaaga agccctggtg gtggcacctc cgtgttcagg 4141 agctgggcct gtcagccccg ctgaccgtgc tcccgaccat cacctgtggg cacaccatcg 4201 agatcctccg ggagaagggc ttcgaccagg cgcccgtggt ggatgaggcg ggggtaatcc 4261 tgggaatggt gacgcttggg aacatgctct cgtccctgct tgccgggaag gtgcagccgt 4321 cagaccaagt tggcaaagtc atctacaagc agttcaaaca gatccgcctc acggacacgc 4381 tgggcaggct ctcgcacatc ctggagatgg accacttcgc cctggtggtg cacgagcaga 4441 tccagtacca cagcaccggg aagtccagtc agcggcagat ggtgttcggg gtggtcaccg 4501 ccattgactt gctgaacttc gtggccgccc aggagcggga ccagaagtga gaattctgca 4561 gtcgacggta ccgcgggccc gggatcccgc gactctagat aattctaccg ggtaggggag 4621 gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc 4681 tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct 4741 ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc 4801 cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt 4861 ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc 4921 caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg 4981 ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg 5041 gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg 5101 ggcctttcga cctgcagccc aagcttacca tgaccgagta caagcccacg gtgcgcctcg 5161 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc 5221 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag 5281 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg 5341 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga 5401 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag 5461 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct 5521 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg 5581 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct 5641 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct 5701 ggtgcatgac ccgcaagccc ggtgcctgac cgcgtctgga acaatcaacc tctggattac 5761 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 5821 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 5881 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 5941 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 6001 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 6061 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 6121 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg 6181 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 6241 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg 6301 agtcggatct ccctttgggc cgcctccccg cctggaatta attctgcagt cgagacctag 6361 aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc 6421 tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac 6481 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag aggggactgg 6541 aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac 6601 aaggctactt ccctgattag cagaactaca caccagggcc aggggtcaga tatccactga 6661 cctttggatg gtgctacaag ctagtaccag ttgagccaga taaggtagaa gaggccaata 6721 aaggagagaa caccagcttg ttacaccctg tgagcctgca tgggatggat gacccggaga 6781 gagaagtgtt agagtggagg tttgacagcc gcctagcatt tcatcacgtg gcccgagagc 6841 tgcatccgga gtacttcaag aactgctgat atcgagcttg ctacaaggga ctttccgctg 6901 gggactttcc agggaggcgt ggcctgggcg ggactgggga gtggcgagcc ctcagatcct 6961 gcatataagc agctgctttt tgcctgtact gggtctctct ggttagacca gatctgagcc 7021 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga 7081 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga 7141 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag 7201 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggccttgaca ttgctagcgt 7261 tttaccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg 7321 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 7381 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 7441 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 7501 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 7561 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 7621 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 7681 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 7741 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 7801 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 7861 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 7921 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 7981 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 8041 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 8101 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg 8161 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 8221 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 8281 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 8341 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 8401 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 8461 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 8521 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 8581 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 8641 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 8701 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 8761 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 8821 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 8881 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 8941 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 9001 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 9061 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 9121 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 9181 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 9241 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 9301 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 9361 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 9421 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atcaacttgt 9481 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag 9541 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg 9601 tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc accccatacc 9661 ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat tcctctgaat 9721 tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagttttta 9781 aagaaattgt atttgttaaa tatgtactac aaacttagta gt //