LOCUS V036353 10119 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V036353 VERSION V036353 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 2827..4134 /label="Fgg(NM_133862)" /note="Fgg(NM_133862)" /gene="Fgg" CDS 4135..4158 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" CDS 4171..4890 /codon_start=1 /note="mammalian codon-optimized" /product="the original enhanced GFP (Yang et al., 1996)" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV LLEFVTAAGITLGMDELYK*" /label="EGFP" promoter 4907..5406 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5427..6026 /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 6040..6628 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6835..7468 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7597..7613) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7621..7637 /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(7645..7675) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7690..7711 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7999..8587) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8758..9618) /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(9619..9723) /gene="bla" /label="AmpR promoter" polyA_signal 9771..9905 /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 gccaccatga gttggtcttt gcagcccccg agtttccttc tttgctgcct gcttttactg 2881 ttctctccaa cgggcctggc atatgtggct accagagata actgttgcat cctagatgag 2941 agattcggta gtttctgccc aaccacctgt ggtatcgcag atttcctgtc ttcttaccaa 3001 accgacgtgg acaatgatct ccgaacgctg gaagacatct tattccgggc tgaaaacaga 3061 accacagaag ccaaagaact gatcaaagca attcaggttt actacaaccc agaccaaccc 3121 ccaaagccag gtatgataga cagtgccact cagaagtcta agaagatggt agaagaaatt 3181 gtgaaatacg aagcgctgct gttaacccac gagacaagca ttcggtattt acaggaaatc 3241 tataattcaa ataaccagaa gatcacaaac ctaaaacaga aggtagccca gcttgaagcc 3301 cagtgccagg agccttgcaa ggactctgtg caaatccatg acacaactgg aaaagattgt 3361 caggaaattg ccaacaaggg cgccaaagaa agtggacttt acttcattcg acctttgaaa 3421 gctaagcagc agttcttagt gtactgtgaa atcgacggat ctggaaatgg ttggaccgta 3481 ttgcagaaga ggattgatgg cagtttggat ttcaagaaga actggattca gtataaagaa 3541 ggatttggac acctgtcgcc tactggcacc acagagtttt ggctgggaaa tgagaagatt 3601 catttgataa gcatgcagtc cacaattcca tatgcactga gaatacagct caaagactgg 3661 aatggcagaa ccagcactgc ggactatgcc atgttcaggg tgggtcccga gtctgacaaa 3721 taccgcctga cctatgccta cttcattggt ggagatgccg gggatgcctt cgacggctac 3781 gatttcggcg atgaccccag tgacaagttt ttcacatccc acaacggcat gcagttcagt 3841 acctgggaca acgacaacga taagtttgaa ggcaactgtg cggaacagga tggatccggc 3901 tggtggatga acaaatgtca cgcaggccac ctcaatggag tttatcacca aggtggtact 3961 tactcaaaat catctactac taatggtttc gacgacggca ttatttgggc gacctggaaa 4021 agccgctggt attctatgaa ggaaaccacc atgaagataa ttcccttcaa cagactctcc 4081 atcggagaag gacagcagca tcacatgggg ggatccaaac aggctggaga cgttgattac 4141 aaggacgacg atgacaaggg atccgccacc atggtgagca agggcgagga gctgttcacc 4201 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg 4261 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc 4321 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag 4381 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc 4441 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc 4501 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac 4561 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac 4621 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac 4681 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc 4741 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa 4801 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc 4861 actctcggca tggacgagct gtacaagtag tctagataat tctaccgggt aggggaggcg 4921 cttttcccaa ggcagtctgg agcatgcgct ttagcagccc cgctgggcac ttggcgctac 4981 acaagtggcc tctggcctcg cacacattcc acatccaccg gtaggcgcca accggctccg 5041 ttctttggtg gccccttcgc gccaccttct actcctcccc tagtcaggaa gttccccccc 5101 gccccgcagc tcgcgtcgtg caggacgtga caaatggaag tagcacgtct cactagtctc 5161 gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag gcctttgggg cagcggccaa 5221 tagcagcttt gctccttcgc tttctgggct cagaggctgg gaaggggtgg gtccgggggc 5281 gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca 5341 ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc atctccgggc 5401 ctttcgacct gcagcccaag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca 5461 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg 5521 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac 5581 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg 5641 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg 5701 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc 5761 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc 5821 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg 5881 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg 5941 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt 6001 gcatgacccg caagcccggt gcctgaccgc gtctggaaca atcaacctct ggattacaaa 6061 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac 6121 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc 6181 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt 6241 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc 6301 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc 6361 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg 6421 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt 6481 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc 6541 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt 6601 cggatctccc tttgggccgc ctccccgcct ggaattaatt ctgcagtcga gacctagaaa 6661 aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag 6721 aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa 6781 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagagg ggactggaag 6841 ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag 6901 gctacttccc tgattagcag aactacacac cagggccagg ggtcagatat ccactgacct 6961 ttggatggtg ctacaagcta gtaccagttg agccagataa ggtagaagag gccaataaag 7021 gagagaacac cagcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag 7081 aagtgttaga gtggaggttt gacagccgcc tagcatttca tcacgtggcc cgagagctgc 7141 atccggagta cttcaagaac tgctgatatc gagcttgcta caagggactt tccgctgggg 7201 actttccagg gaggcgtggc ctgggcggga ctggggagtg gcgagccctc agatcctgca 7261 tataagcagc tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg 7321 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg 7381 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc 7441 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat 7501 ttataacttg caaagaaatg aatatcagag agtgagaggc cttgacattg ctagcgtttt 7561 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa 7621 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg 7681 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca 7741 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg 7801 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 7861 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 7921 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 7981 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 8041 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 8101 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 8161 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 8221 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 8281 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 8341 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 8401 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 8461 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 8521 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 8581 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 8641 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 8701 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 8761 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 8821 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 8881 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca 8941 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 9001 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 9061 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 9121 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 9181 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 9241 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 9301 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 9361 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat 9421 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 9481 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 9541 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 9601 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 9661 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 9721 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc aacttgttta 9781 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 9841 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct 9901 ggatcaactg gataactcaa gctaaccaaa atcatcccaa acttcccacc ccatacccta 9961 ttaccactgc caattacctg tggtttcatt tactctaaac ctgtgattcc tctgaattat 10021 tttcatttta aagaaattgt atttgttaaa tatgtactac aaacttagta gtttttaaag 10081 aaattgtatt tgttaaatat gtactacaaa cttagtagt //