LOCUS V036484 8969 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V036484 VERSION V036484 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..3695 /label="CD274(NM_014143)" /note="CD274(NM_014143)" /gene="CD274" CDS 3702..3725 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" promoter 3757..4256 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4277..4876 /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 4890..5478 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5685..6318 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6447..6463) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6471..6487 /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(6495..6525) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6540..6561 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6849..7437) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7608..8468) /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(8469..8573) /gene="bla" /label="AmpR promoter" polyA_signal 8621..8755 /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 ccaccatgag gatatttgct gtctttatat tcatgaccta ctggcatttg ctgaacgcat 2881 ttactgtcac ggttcccaag gacctatatg tggtagagta tggtagcaat atgacaattg 2941 aatgcaaatt cccagtagaa aaacaattag acctggctgc actaattgtc tattgggaaa 3001 tggaggataa gaacattatt caatttgtgc atggagagga agacctgaag gttcagcata 3061 gtagctacag acagagggcc cggctgttga aggaccagct ctccctggga aatgctgcac 3121 ttcagatcac agatgtgaaa ttgcaggatg caggggtgta ccgctgcatg atcagctatg 3181 gtggtgccga ctacaagcga attactgtga aagtcaatgc cccatacaac aaaatcaacc 3241 aaagaatttt ggttgtggat ccagtcacct ctgaacatga actgacatgt caggctgagg 3301 gctaccccaa ggccgaagtc atctggacaa gcagtgacca tcaagtcctg agtggtaaga 3361 ccaccaccac caattccaag agagaggaga agcttttcaa tgtgaccagc acactgagaa 3421 tcaacacaac aactaatgag attttctact gcacttttag gagattagat cctgaggaaa 3481 accatacagc tgaattggtc atcccagaac tacctctggc acatcctcca aatgaaagga 3541 ctcacttggt aattctggga gccatcttat tatgccttgg tgtagcactg acattcatct 3601 tccgtttaag aaaagggaga atgatggatg tgaaaaaatg tggcatccaa gatacaaact 3661 caaagaagca aagtgataca catttggagg agacggaatt cgattacaag gatgacgacg 3721 ataagtgagg atcccgcgac tctagataat tctaccgggt aggggaggcg cttttcccaa 3781 ggcagtctgg agcatgcgct ttagcagccc cgctgggcac ttggcgctac acaagtggcc 3841 tctggcctcg cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg 3901 gccccttcgc gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc 3961 tcgcgtcgtg caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg 4021 acagcaccgc tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt 4081 gctccttcgc tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg 4141 gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc 4201 ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc atctccgggc ctttcgacct 4261 gcagcccaag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga 4321 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca 4381 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac 4441 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt 4501 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat 4561 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc 4621 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca 4681 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg 4741 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg 4801 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg 4861 caagcccggt gcctgaccgc gtctggaaca atcaacctct ggattacaaa atttgtgaaa 4921 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa 4981 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat 5041 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt 5101 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc 5161 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc 5221 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg 5281 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga 5341 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc 5401 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc 5461 tttgggccgc ctccccgcct ggaattaatt ctgcagtcga gacctagaaa aacatggagc 5521 aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga 5581 ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa 5641 ggcagctgta gatcttagcc actttttaaa agaaaagagg ggactggaag ggctaattca 5701 ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag gctacttccc 5761 tgattagcag aactacacac cagggccagg ggtcagatat ccactgacct ttggatggtg 5821 ctacaagcta gtaccagttg agccagataa ggtagaagag gccaataaag gagagaacac 5881 cagcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag aagtgttaga 5941 gtggaggttt gacagccgcc tagcatttca tcacgtggcc cgagagctgc atccggagta 6001 cttcaagaac tgctgatatc gagcttgcta caagggactt tccgctgggg actttccagg 6061 gaggcgtggc ctgggcggga ctggggagtg gcgagccctc agatcctgca tataagcagc 6121 tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg 6181 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag 6241 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag 6301 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg 6361 caaagaaatg aatatcagag agtgagaggc cttgacattg ctagcgtttt accgtcgacc 6421 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg 6481 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa 6541 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac 6601 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt 6661 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga 6721 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca 6781 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg 6841 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt 6901 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc 6961 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct 7021 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc 7081 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta 7141 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca 7201 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag 7261 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag 7321 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt 7381 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa 7441 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg 7501 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga 7561 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta 7621 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc 7681 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg 7741 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga 7801 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt 7861 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt 7921 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc 7981 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc 8041 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca 8101 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag 8161 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg 8221 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa 8281 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa 8341 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga 8401 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga 8461 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg 8521 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt 8581 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc aacttgttta ttgcagctta 8641 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 8701 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaactg 8761 gataactcaa gctaaccaaa atcatcccaa acttcccacc ccatacccta ttaccactgc 8821 caattacctg tggtttcatt tactctaaac ctgtgattcc tctgaattat tttcatttta 8881 aagaaattgt atttgttaaa tatgtactac aaacttagta gtttttaaag aaattgtatt 8941 tgttaaatat gtactacaaa cttagtagt //