LOCUS V033429 9610 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V033429 VERSION V033429 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..4306 /label="PDIA6(NM_001282705)" /note="PDIA6(NM_001282705)" /gene="PDIA6" CDS 4313..4378 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4398..4897 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4918..5517 /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 5531..6119 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6326..6959 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7088..7104) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7112..7128 /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(7136..7166) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7181..7202 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7490..8078) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8249..9109) /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(9110..9214) /gene="bla" /label="AmpR promoter" polyA_signal 9262..9396 /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 ccatgaggat aatcacagcc cctgcctcca aagtgtcccg 2881 aggatcaaat gagttgatga tacttgcacg gcgctcggat cgtggctcac caacttctcc 2941 tgcccactcc ctatcaagga aaagtcccat aatgtatcca tcaaccacca tggctaatgc 3001 acccggtctg gtgagctgta ccttctttct ggcagtgaat ggtctgtatt cctctagtga 3061 tgatgtgatc gaattaactc catcgaattt caaccgagaa gttattcaga gtgatagttt 3121 gtggcttgta gaattctatg ctccatggtg tggtcactgt caaagattaa caccagaatg 3181 gaagaaagca gcaactgcat taaaagatgt tgtcaaagtt ggtgcagttg atgcagataa 3241 gcatcattcc ctaggaggtc agtatggtgt tcagggattt cctaccatta agatttttgg 3301 atccaacaaa aacagaccag aagattacca aggtggcaga actggtgaag ccattgtaga 3361 tgctgcgctg agtgctctgc gccagctcgt gaaggatcgc ctcgggggac ggagcggagg 3421 atacagttct ggaaaacaag gcagaagtga tagttcaagt aagaaggatg tgattgagct 3481 gacagacgac agctttgata agaatgttct ggacagtgaa gatgtttgga tggttgagtt 3541 ctatgctcct tggtgtggac actgcaaaaa cctagagcca gagtgggctg ccgcagcttc 3601 agaagtaaaa gagcagacga aaggaaaagt gaaactggca gctgtggatg ctacagtcaa 3661 tcaggttctg gcctcccgat acgggattag aggatttcct acaatcaaga tatttcagaa 3721 aggcgagtct cctgtggatt atgacggtgg gcggacaaga tccgacatcg tgtcccgggc 3781 ccttgatttg ttttctgata acgccccacc tcctgagctg cttgagatta tcaacgagga 3841 cattgccaag aggacgtgtg aggagcacca gctctgtgtt gtggctgtgc tgccccatat 3901 ccttgatact ggagctgcag gcagaaattc ttatctggaa gttcttctga agttggcaga 3961 caaatacaaa aagaaaatgt gggggtggct gtggacagaa gctggagccc agtctgaact 4021 tgagaccgcg ttggggattg gagggtttgg gtaccccgcc atggccgcca tcaatgcacg 4081 caagatgaaa tttgctctgc taaaaggctc cttcagtgag caaggcatca acgagtttct 4141 cagggagctc tcttttgggc gtggctccac ggcacctgta ggaggcgggg ctttccctac 4201 catcgttgag agagagcctt gggacggcag ggatggcgag cttcccgtgg aggatgacat 4261 tgacctcagt gatgtggagc ttgatgactt agggaaagat gagttgggat ccgactacaa 4321 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta 4381 gtctagataa ttctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc 4441 tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc 4501 cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc 4561 tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg 4621 acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg ctgagcaatg 4681 gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc 4741 tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc 4801 gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc 4861 cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcagcccaa gcttaccatg 4921 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc 4981 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc 5041 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc 5101 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc 5161 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc 5221 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc 5281 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc 5341 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag 5401 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac 5461 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaccg 5521 cgtctggaac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa 5581 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat 5641 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta 5701 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc 5761 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt 5821 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg 5881 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc 5941 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc 6001 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct 6061 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc 6121 tggaattaat tctgcagtcg agacctagaa aaacatggag caatcacaag tagcaataca 6181 gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt 6241 ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc 6301 cactttttaa aagaaaagag gggactggaa gggctaattc actcccaacg aagacaagat 6361 atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattagca gaactacaca 6421 ccagggccag gggtcagata tccactgacc tttggatggt gctacaagct agtaccagtt 6481 gagccagata aggtagaaga ggccaataaa ggagagaaca ccagcttgtt acaccctgtg 6541 agcctgcatg ggatggatga cccggagaga gaagtgttag agtggaggtt tgacagccgc 6601 ctagcatttc atcacgtggc ccgagagctg catccggagt acttcaagaa ctgctgatat 6661 cgagcttgct acaagggact ttccgctggg gactttccag ggaggcgtgg cctgggcggg 6721 actggggagt ggcgagccct cagatcctgc atataagcag ctgctttttg cctgtactgg 6781 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact 6841 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg 6901 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag 6961 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga 7021 gagtgagagg ccttgacatt gctagcgttt taccgtcgac ctctagctag agcttggcgt 7081 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 7141 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 7201 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 7261 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 7321 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 7381 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 7441 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 7501 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 7561 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 7621 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 7681 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 7741 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 7801 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 7861 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 7921 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 7981 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 8041 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 8101 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 8161 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 8221 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 8281 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 8341 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 8401 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 8461 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 8521 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 8581 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 8641 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 8701 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 8761 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 8821 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 8881 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 8941 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 9001 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 9061 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 9121 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 9181 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 9241 acgtcgacgg atcgggagat caacttgttt attgcagctt ataatggtta caaataaagc 9301 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 9361 tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa 9421 aatcatccca aacttcccac cccataccct attaccactg ccaattacct gtggtttcat 9481 ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa 9541 atatgtacta caaacttagt agtttttaaa gaaattgtat ttgttaaata tgtactacaa 9601 acttagtagt //