LOCUS V026914 9546 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V026914 VERSION V026914 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 2821..2830 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 2830..2853 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" CDS 2854..4302 /label="DDX6(NM_004397)" /note="DDX6(NM_004397)" /gene="DDX6" promoter 4334..4833 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4854..5453 /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 5467..6055 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6262..6895 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7024..7040) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7048..7064 /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(7072..7102) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7117..7138 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7426..8014) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8185..9045) /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(9046..9150) /gene="bla" /label="AmpR promoter" polyA_signal 9198..9332 /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 gccgccatgg actacaagga cgacgatgac aagatgagca cggccagaac agagaaccct 2881 gttataatgg gtctgtccag tcaaaatggt cagctgagag gccctgtgaa acccactggt 2941 ggccctggag gagggggcac acagacacag caacagatga accagctgaa aaacaccaac 3001 acaatcaata atggcactca gcagcaagca cagagtatga ccaccactat taaacctggt 3061 gatgactgga aaaagacttt aaaactccct ccaaaggatc taagaatcaa aacttcggat 3121 gtgacctcca caaaaggaaa tgagtttgaa gattactgtt tgaaacggga gttactgatg 3181 ggaatttttg aaatgggctg ggaaaagcca tctcctattc aggaggagag cattcccatt 3241 gctttatctg gtagggatat cttagctaga gcaaaaaatg gaacaggcaa gagcggtgcc 3301 tacctcattc ccttacttga acggctagac ctgaagaagg acaatataca agcaatggtg 3361 attgttccca ctagagaact tgctctacag gtcagtcaaa tttgcatcca ggtcagcaaa 3421 cacatgggag gggccaaagt gatggcaacc acaggaggaa ccaatttacg agatgacata 3481 atgaggcttg atgatacagt gcacgtggtg attgctaccc ctgggagaat cctggatctt 3541 attaagaaag gagtagcaaa ggttgatcat gtccagatga tagtattgga tgaggcagat 3601 aagttgctgt cacaggattt tgtgcagata atggaggata ttattctcac gctacctaaa 3661 aacaggcaga ttttactata ttccgctact ttccctctta gtgtacagaa gttcatgaat 3721 tcccatttgc agaaacccta tgagattaac ctgatggagg aactaactct gaagggagta 3781 acccagtact acgcatatgt aactgagcgc caaaaagtac actgcctcaa cacacttttc 3841 tccaggcttc agataaacca gtcgatcatt ttctgtaact cctctcagcg agttgaattg 3901 ctagccaaga agatttctca actgggttat tcttgcttct atattcatgc taaaatgagg 3961 caggaacatc gaaatcgtgt atttcatgat ttccgaaatg gcttatgccg caatcttgtt 4021 tgcactgatc tgtttacccg aggtattgat atacaagctg tgaatgtggt aataaacttt 4081 gatttcccaa agctggcaga gacctatctc catcgtattg gaagatcagg tcgctttggt 4141 catcttggct tagccatcaa cttgatcaca tatgatgatc gcttcaacct gaaaagtatt 4201 gaggagcagc tgggaacaga aattaaacct attccgagca acattgataa gagcctgtat 4261 gtggcagaat accacagcga gcctgtagaa gatgagaaac cttaaggatc ccgcgactct 4321 agataattct accgggtagg ggaggcgctt ttcccaaggc agtctggagc atgcgcttta 4381 gcagccccgc tgggcacttg gcgctacaca agtggcctct ggcctcgcac acattccaca 4441 tccaccggta ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc accttctact 4501 cctcccctag tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa 4561 atggaagtag cacgtctcac tagtctcgtg cagatggaca gcaccgctga gcaatggaag 4621 cgggtaggcc tttggggcag cggccaatag cagctttgct ccttcgcttt ctgggctcag 4681 aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg cggggcgggc 4741 gcccgaaggt cctccggagg cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg 4801 ctgttctcct cttcctcatc tccgggcctt tcgacctgca gcccaagctt accatgaccg 4861 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc 4921 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca 4981 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca 5041 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg 5101 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc 5161 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt 5221 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg 5281 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct 5341 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg 5401 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgaccgcgtc 5461 tggaacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat 5521 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct 5581 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag 5641 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc 5701 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc 5761 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct 5821 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg 5881 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg 5941 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg 6001 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga 6061 attaattctg cagtcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag 6121 ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag 6181 tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact 6241 ttttaaaaga aaagagggga ctggaagggc taattcactc ccaacgaaga caagatatcc 6301 ttgatctgtg gatctaccac acacaaggct acttccctga ttagcagaac tacacaccag 6361 ggccaggggt cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc 6421 cagataaggt agaagaggcc aataaaggag agaacaccag cttgttacac cctgtgagcc 6481 tgcatgggat ggatgacccg gagagagaag tgttagagtg gaggtttgac agccgcctag 6541 catttcatca cgtggcccga gagctgcatc cggagtactt caagaactgc tgatatcgag 6601 cttgctacaa gggactttcc gctggggact ttccagggag gcgtggcctg ggcgggactg 6661 gggagtggcg agccctcaga tcctgcatat aagcagctgc tttttgcctg tactgggtct 6721 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 6781 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 6841 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt 6901 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt 6961 gagaggcctt gacattgcta gcgttttacc gtcgacctct agctagagct tggcgtaatc 7021 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 7081 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 7141 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 7201 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 7261 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 7321 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 7381 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 7441 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 7501 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 7561 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 7621 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 7681 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 7741 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 7801 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 7861 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 7921 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 7981 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 8041 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 8101 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 8161 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 8221 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 8281 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 8341 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 8401 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 8461 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 8521 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 8581 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 8641 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 8701 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 8761 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 8821 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 8881 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 8941 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 9001 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 9061 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 9121 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 9181 cgacggatcg ggagatcaac ttgtttattg cagcttataa tggttacaaa taaagcaata 9241 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 9301 aactcatcaa tgtatcttat catgtctgga tcaactggat aactcaagct aaccaaaatc 9361 atcccaaact tcccacccca taccctatta ccactgccaa ttacctgtgg tttcatttac 9421 tctaaacctg tgattcctct gaattatttt cattttaaag aaattgtatt tgttaaatat 9481 gtactacaaa cttagtagtt tttaaagaaa ttgtatttgt taaatatgta ctacaaactt 9541 agtagt //