LOCUS V043522 10330 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V043522 VERSION V043522 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..5005 /label="LARGE2(NM_152312)" /note="LARGE2(NM_152312)" /gene="LARGE2" CDS 5021..5086 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 5118..5617 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5638..6237 /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 6251..6839 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 7046..7679 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7808..7824) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7832..7848 /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(7856..7886) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7901..7922 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8210..8798) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8969..9829) /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(9830..9934) /gene="bla" /label="AmpR promoter" polyA_signal 9982..10116 /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 ccatgctgcc ccgagggcgc ccccgggcgc tgggggccgc 2881 cgcgctgttg ctgctgctgc tgctgctcgg attcctcctg ttcggtgggg acctggggtg 2941 tgagcgccgc gagcctggcg ggcgagcggg ggccccggga tgcttccccg gcccgctcat 3001 gccacgtgtc cccccagacg ggaggctgcg gagagccgcc gccctcgacg gagacccggg 3061 ggccggcccc ggggaccaca accgctccga ctgcggcccg cagccgccgc cgccgcccaa 3121 gtgcgagctc ttgcatgtgg ccatcgtgtg tgcggggcat aactccagcc gagacgtcat 3181 caccctggtg aagtccatgc tcttctacag gaaaaatcca ctgcacctcc acttggtgac 3241 tgacgccgtg gccagaaaca tcctggagac gctcttccac acatggatgg tgcctgctgt 3301 ccgtgtcagc ttttatcatg ccgaccagct caagccccag gtctcctgga tccccaacaa 3361 gcactactcc ggcctctatg ggctaatgaa gctggtgctg cccagtgcct tgcctgctga 3421 gctggcccgc gtcattgtcc tggacacgga tgtcaccttc gcctctgaca tctcggagct 3481 ctgggccctc tttgctcact tttctgacac gcaggcgatc ggtcttgtgg agaaccagag 3541 tgactggtac ctgggcaacc tctggaagaa ccacaggccc tggcctgcct tgggccgggg 3601 atttaacaca ggtgtgatcc tgctgcggct ggaccggctc cggcaggctg gctgggagca 3661 gatgtggagg ctgacagcca ggcgggagct ccttagcctg cctgccacct cactggctga 3721 ccaggacatc ttcaacgctg tgatcaagga gcacccgggg ctagtgcagc gtctgccttg 3781 tgtctggaat gtgcagctgt cagatcacac actggccgag cgctgctact ctgaggcgtc 3841 tgacctcaag gtgatccact ggaactcacc aaagaagctt cgggtgaaga acaagcatgt 3901 ggaattcttc cgcaatttct acctgacctt cctggagtac gatgggaacc tgctgcggag 3961 agagctcttt gtgtgcccca gccagccccc acctggtgct gagcagttgc agcaggccct 4021 ggcacaactg gacgaggaag acccctgctt tgagttccgg cagcagcagc tcactgtgca 4081 ccgtgtgcat gtcactttcc tgccccatga accgccaccc ccccggcctc acgatgtcac 4141 ccttgtggcc cagctgtcca tggaccggct gcagatgttg gaagccctgt gcaggcactg 4201 gcctggcccc atgagcctgg ccttgtacct gacagacgca gaagctcagc agttcctgca 4261 tttcgtcgag gcctcaccag tgcttgctgc ccggcaggac gtggcctacc atgtggtgta 4321 ccgtgagggg cccctatacc ccgtcaacca gcttcgcaac gtggccttgg cccaggccct 4381 cacgccttac gtcttcctca gtgacattga cttcctgcct gcctattctc tctacgacta 4441 cctcagggcc tccattgagc agctggggct gggcagccgg cgcaaggcag cactggtggt 4501 gccggcattc gagaccctgc gctaccgctt cagcttcccc cattccaagg tggagctgtt 4561 ggccttgctg gatgcgggca ctctctacac cttcaggtac cacgagtggc cccgaggcca 4621 cgcacccaca gactatgccc gctggcggga ggctcaggcc ccgtaccgtg tgcaatgggc 4681 ggccaactat gaaccctacg tggtggtgcc acgagactgt ccccgctatg atcctcgctt 4741 tgtgggcttc ggctggaaca aagtggccca cattgtggag ctggatgccc aggaatatga 4801 gctcctggtg ctgcccgagg ccttcaccat ccatctgccc cacgctccaa gcctggacat 4861 ctcccgcttc cgctccagcc ccacctatcg tgactgcctc caggccctca aggacgaatt 4921 ccaccaggac ttgtcccgcc accatggggc tgctgccctc aaatacctcc cagccctgca 4981 gcagccccag agccctgccc gaggcggtgg aggcggatca gactacaaag accatgacgg 5041 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagg gatcccgcga 5101 ctctagataa ttctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc 5161 tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc 5221 cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc 5281 tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg 5341 acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg ctgagcaatg 5401 gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc 5461 tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc 5521 gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc 5581 cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcagcccaa gcttaccatg 5641 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc 5701 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc 5761 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc 5821 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc 5881 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc 5941 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc 6001 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc 6061 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag 6121 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac 6181 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaccg 6241 cgtctggaac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa 6301 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat 6361 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta 6421 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc 6481 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt 6541 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg 6601 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc 6661 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc 6721 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct 6781 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc 6841 tggaattaat tctgcagtcg agacctagaa aaacatggag caatcacaag tagcaataca 6901 gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt 6961 ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc 7021 cactttttaa aagaaaagag gggactggaa gggctaattc actcccaacg aagacaagat 7081 atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattagca gaactacaca 7141 ccagggccag gggtcagata tccactgacc tttggatggt gctacaagct agtaccagtt 7201 gagccagata aggtagaaga ggccaataaa ggagagaaca ccagcttgtt acaccctgtg 7261 agcctgcatg ggatggatga cccggagaga gaagtgttag agtggaggtt tgacagccgc 7321 ctagcatttc atcacgtggc ccgagagctg catccggagt acttcaagaa ctgctgatat 7381 cgagcttgct acaagggact ttccgctggg gactttccag ggaggcgtgg cctgggcggg 7441 actggggagt ggcgagccct cagatcctgc atataagcag ctgctttttg cctgtactgg 7501 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact 7561 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg 7621 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag 7681 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga 7741 gagtgagagg ccttgacatt gctagcgttt taccgtcgac ctctagctag agcttggcgt 7801 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 7861 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 7921 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 7981 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 8041 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 8101 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 8161 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 8221 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 8281 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 8341 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 8401 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 8461 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 8521 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 8581 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 8641 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 8701 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 8761 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 8821 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 8881 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 8941 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 9001 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 9061 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 9121 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 9181 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 9241 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 9301 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 9361 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 9421 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 9481 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 9541 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 9601 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 9661 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 9721 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 9781 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 9841 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 9901 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 9961 acgtcgacgg atcgggagat caacttgttt attgcagctt ataatggtta caaataaagc 10021 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 10081 tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa 10141 aatcatccca aacttcccac cccataccct attaccactg ccaattacct gtggtttcat 10201 ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa 10261 atatgtacta caaacttagt agtttttaaa gaaattgtat ttgttaaata tgtactacaa 10321 acttagtagt //