LOCUS V022116 9310 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022116 VERSION V022116 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 2826..4049 /label="Map2k4(NM_001316367)" /note="Map2k4(NM_001316367)" /gene="Map2k4" CDS 4050..4067 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" promoter 4098..4597 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4618..5217 /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 5231..5819 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6026..6659 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6788..6804) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6812..6828 /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(6836..6866) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6881..6902 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7190..7778) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7949..8809) /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(8810..8914) /gene="bla" /label="AmpR promoter" polyA_signal 8962..9096 /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 ccaccatggc ggctccgagc ccgagcggcg gcggcggctc cgggggcggt ggcggcaccc 2881 cggggcccat cggacctcca gcttctggcc acccggcggt cagcagcatg cagggctttc 2941 agataaactt ctgtgaaaag gcacaaagta agcgcaaagc actgaagttg aattttgcaa 3001 atccacctgt caaatcgaca gcacggttta ctctgaatcc taatactaca ggagtccaga 3061 acccacacat agagagactg agaacacaca gcattgagtc atcaggaaaa ctgaagatct 3121 cccctgaaca acactgggat ttcactgcag aggacttgaa agaccttgga gaaattggac 3181 gaggagctta tggttctgtc aacaaaatgg tccacaaacc aagtgggcag ataatggcag 3241 ttaaaagaat tcggtcaact gtggatgaaa aagaacaaaa acaacttctc atggatttgg 3301 atgtagtaat gcggagtagt gattgcccat acattgttca gttctatggt gcactcttca 3361 gagagggcga ctgttggatc tgtatggagc tcatgtctac ctcgttcgat aagttttaca 3421 aatatgtata tagtgtgtta gatgacgtta ttccggaaga gatcttaggc aaaatcactt 3481 tagcaactgt gaaagcacta aaccacttaa aagaaaactt gaaaattatt cacagagaca 3541 tcaaaccttc caatattctt ctggacagaa gtggaaatat aaagctctgt gatttcggca 3601 tcagtggaca gcttgtggac tctattgcca agacaagaga tgctgggtgt aggccgtata 3661 tggcacctga aagaatagac ccaagtgcat caagacaagg gtatgatgtc cgctctgatg 3721 tctggagttt ggggatcaca ttgtacgagt tggccacagg ccgatttcct tatccaaagt 3781 ggaatagtgt atttgatcag ctaacacaag tggtgaaagg agaccctccg cagctgagta 3841 attctgaaga aagggagttc tcccccagtt tcatcaactt tgtcaacttg tgccttacga 3901 aggatgaatc caaaaggcca aagtataaag agcttctgaa acatcccttt attttgatgt 3961 atgaagcacg tactgtagag gtcgcatgct atgtttgtaa aatcctggat cagatgccag 4021 ccactcccag ctcgcccatg tatgtcgacc accatcacca tcaccattag gatcccgcga 4081 ctctagataa ttctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc 4141 tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc 4201 cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc 4261 tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg 4321 acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg ctgagcaatg 4381 gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc 4441 tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc 4501 gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc 4561 cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcagcccaa gcttaccatg 4621 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc 4681 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc 4741 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc 4801 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc 4861 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc 4921 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc 4981 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc 5041 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag 5101 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac 5161 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaccg 5221 cgtctggaac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa 5281 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat 5341 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta 5401 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc 5461 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt 5521 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg 5581 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc 5641 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc 5701 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct 5761 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc 5821 tggaattaat tctgcagtcg agacctagaa aaacatggag caatcacaag tagcaataca 5881 gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt 5941 ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc 6001 cactttttaa aagaaaagag gggactggaa gggctaattc actcccaacg aagacaagat 6061 atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattagca gaactacaca 6121 ccagggccag gggtcagata tccactgacc tttggatggt gctacaagct agtaccagtt 6181 gagccagata aggtagaaga ggccaataaa ggagagaaca ccagcttgtt acaccctgtg 6241 agcctgcatg ggatggatga cccggagaga gaagtgttag agtggaggtt tgacagccgc 6301 ctagcatttc atcacgtggc ccgagagctg catccggagt acttcaagaa ctgctgatat 6361 cgagcttgct acaagggact ttccgctggg gactttccag ggaggcgtgg cctgggcggg 6421 actggggagt ggcgagccct cagatcctgc atataagcag ctgctttttg cctgtactgg 6481 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact 6541 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg 6601 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag 6661 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga 6721 gagtgagagg ccttgacatt gctagcgttt taccgtcgac ctctagctag agcttggcgt 6781 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 6841 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 6901 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 6961 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 7021 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 7081 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 7141 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 7201 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 7261 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 7321 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 7381 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 7441 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 7501 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 7561 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 7621 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 7681 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 7741 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 7801 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 7861 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 7921 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 7981 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 8041 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 8101 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 8161 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 8221 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 8281 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 8341 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 8401 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 8461 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 8521 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 8581 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 8641 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 8701 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 8761 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 8821 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 8881 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 8941 acgtcgacgg atcgggagat caacttgttt attgcagctt ataatggtta caaataaagc 9001 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 9061 tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa 9121 aatcatccca aacttcccac cccataccct attaccactg ccaattacct gtggtttcat 9181 ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa 9241 atatgtacta caaacttagt agtttttaaa gaaattgtat ttgttaaata tgtactacaa 9301 acttagtagt //