LOCUS V028461 8950 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V028461 VERSION V028461 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..3625 /label="Rchy1(NM_001007618)" /note="Rchy1(NM_001007618)" /gene="Rchy1" CDS 3632..3658 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3662..3688 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3692..3718 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" promoter 3738..4237 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4258..4857 /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 4871..5459 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5666..6299 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6428..6444) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6452..6468 /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(6476..6506) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6521..6542 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6830..7418) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7589..8449) /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(8450..8554) /gene="bla" /label="AmpR promoter" polyA_signal 8602..8736 /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 ccatggcggc gacggcgcag gaagatggcg tccgcagccc 2881 ggctccaggg ccgcggggtt gcgagcacta tgacagagcc tgtctcctca aggcaccttg 2941 ttgtgacaag ctctatacat gccggctatg tcacgacacc cacgaagatc atcagctgga 3001 ccgcttcaaa gtaaaggaag tgcagtgcat caactgtgaa aaactccagc atgcccagca 3061 gacttgtgag gactgcagca cactgtttgg agaatattac tgcagtattt gccacctgtt 3121 tgacaaagac aagaagcagt atcactgtga gagctgtggg atttgtagga ttggtccaaa 3181 ggaagatttt tttcattgct tgaaatgtaa cttatgccta gccatgactc ttcgaggaaa 3241 acacaagtgt attgaaaatg tttcccggca gaattgtcca atatgcttgg aggacattca 3301 tacctcccgt gtcgttgctc atgtcttgcc atgcgggcat ctcctacata gaacgtgtta 3361 tgaagaaatg ttgaaagaag gttacagatg cccattgtgt atgcattctg ccctagacat 3421 gactcggtac tggagacaat tagacattga ggtagcacag actcctatgc catccgaata 3481 ccagaacgtg accgttgata ttctctgcaa tgactgtaat ggacgatcca cagtccagtt 3541 ccatatctta ggcatgaaat gtaagctttg tgactcctac aatactgctc aggctggagg 3601 ccgcacagtg tccatggatg agcagggatc ctacccatac gacgtacctg attatgcaac 3661 ttatccttat gatgtacctg attatgctag ctatccttat gatgtcccag actacgcata 3721 gtctagataa ttctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc 3781 tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc 3841 cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc 3901 tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg 3961 acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg ctgagcaatg 4021 gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc 4081 tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc 4141 gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc 4201 cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcagcccaa gcttaccatg 4261 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc 4321 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc 4381 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc 4441 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc 4501 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc 4561 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc 4621 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc 4681 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag 4741 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac 4801 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaccg 4861 cgtctggaac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa 4921 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat 4981 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta 5041 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc 5101 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt 5161 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg 5221 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc 5281 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc 5341 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct 5401 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc 5461 tggaattaat tctgcagtcg agacctagaa aaacatggag caatcacaag tagcaataca 5521 gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt 5581 ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc 5641 cactttttaa aagaaaagag gggactggaa gggctaattc actcccaacg aagacaagat 5701 atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattagca gaactacaca 5761 ccagggccag gggtcagata tccactgacc tttggatggt gctacaagct agtaccagtt 5821 gagccagata aggtagaaga ggccaataaa ggagagaaca ccagcttgtt acaccctgtg 5881 agcctgcatg ggatggatga cccggagaga gaagtgttag agtggaggtt tgacagccgc 5941 ctagcatttc atcacgtggc ccgagagctg catccggagt acttcaagaa ctgctgatat 6001 cgagcttgct acaagggact ttccgctggg gactttccag ggaggcgtgg cctgggcggg 6061 actggggagt ggcgagccct cagatcctgc atataagcag ctgctttttg cctgtactgg 6121 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact 6181 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg 6241 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag 6301 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga 6361 gagtgagagg ccttgacatt gctagcgttt taccgtcgac ctctagctag agcttggcgt 6421 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 6481 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 6541 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 6601 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 6661 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 6721 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 6781 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 6841 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 6901 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 6961 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 7021 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 7081 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 7141 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 7201 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 7261 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 7321 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 7381 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 7441 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 7501 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 7561 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 7621 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 7681 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 7741 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 7801 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 7861 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 7921 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 7981 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 8041 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 8101 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 8161 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 8221 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 8281 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 8341 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 8401 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 8461 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 8521 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 8581 acgtcgacgg atcgggagat caacttgttt attgcagctt ataatggtta caaataaagc 8641 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 8701 tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa 8761 aatcatccca aacttcccac cccataccct attaccactg ccaattacct gtggtttcat 8821 ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa 8881 atatgtacta caaacttagt agtttttaaa gaaattgtat ttgttaaata tgtactacaa 8941 acttagtagt //