LOCUS V030336 11026 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V030336 VERSION V030336 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..5722 /label="OPA1(NM_015560)" /note="OPA1(NM_015560)" /gene="OPA1" CDS 5729..5794 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 5814..6313 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 6334..6933 /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 6947..7535 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 7742..8375 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(8504..8520) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 8528..8544 /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(8552..8582) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 8597..8618 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8906..9494) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(9665..10525) /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(10526..10630) /gene="bla" /label="AmpR promoter" polyA_signal 10678..10812 /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 ccatgtggcg actacgtcgg gccgctgtgg cctgtgaggt 2881 ctgccagtct ttagtgaaac acagctctgg aataaaagga agtttaccac tacaaaaact 2941 acatctggtt tcacgaagca tttatcattc acatcatcct accttaaagc ttcaacgacc 3001 ccaattaagg acatcctttc agcagttctc ttctctgaca aaccttcctt tacgtaaact 3061 gaaattctct ccaattaaat atggctacca gcctcgcagg aatttttggc cagcaagatt 3121 agctacgaga ctcttaaaac ttcgctatct catactagga tcggctgttg ggggtggcta 3181 cacagccaaa aagacttttg atcagtggaa agatatgata ccggacctta gtgaatataa 3241 atggattgtg cctgacattg tgtgggaaat tgatgagtat atcgattttg agaaaattag 3301 aaaagccctt cctagttcag aagaccttgt aaagttagca ccagactttg acaagattgt 3361 tgaaagcctt agcttattga aggacttttt tacctcaggt tctccggaag aaacggcgtt 3421 tagagcaaca gatcgtggat ctgaaagtga caagcatttt agaaaggtgt cagacaaaga 3481 gaaaattgac caacttcagg aagaacttct gcacactcag ttgaagtatc agagaatctt 3541 ggaacgatta gaaaaggaga acaaagaatt gagaaaatta gtattgcaga aagatgacaa 3601 aggcattcat catagaaagc ttaagaaatc tttgattgac atgtattctg aagttcttga 3661 tgttctctct gattatgatg ccagttataa tacgcaagat catctgccac gggttgttgt 3721 ggttggagat cagagtgctg gaaagactag tgtgttggaa atgattgccc aagctcgaat 3781 attcccaaga ggatctgggg agatgatgac acgttctcca gttaaggtga ctctgagtga 3841 aggtcctcac catgtggccc tatttaaaga tagttctcgg gagtttgatc ttaccaaaga 3901 agaagatctt gcagcattaa gacatgaaat agaacttcga atgaggaaaa atgtgaaaga 3961 aggctgtacc gttagccctg agaccatatc cttaaatgta aaaggccctg gactacagag 4021 gatggtgctt gttgacttac caggtgtgat taatactgtg acatcaggca tggctcctga 4081 cacaaaggaa actattttca gtatcagcaa agcttacatg cagaatccta atgccatcat 4141 actgtgtatt caagatggat ctgtggatgc tgaacgcagt attgttacag acttggtcag 4201 tcaaatggac cctcatggaa ggagaaccat attcgttttg accaaagtag acctggcaga 4261 gaaaaatgta gccagtccaa gcaggattca gcagataatt gaaggaaagc tcttcccaat 4321 gaaagcttta ggttattttg ctgttgtaac aggaaaaggg aacagctctg aaagcattga 4381 agctataaga gaatatgaag aagagttttt tcagaattca aagctcctaa agacaagcat 4441 gctaaaggca caccaagtga ctacaagaaa tttaagcctt gcagtatcag actgcttttg 4501 gaaaatggta cgagagtctg ttgaacaaca ggctgatagt ttcaaagcaa cacgttttaa 4561 ccttgaaact gaatggaaga ataactatcc tcgcctgcgg gaacttgacc ggaatgaact 4621 atttgaaaaa gctaaaaatg aaatccttga tgaagttatc agtctgagcc aggttacacc 4681 aaaacattgg gaggaaatcc ttcaacaatc tttgtgggaa agagtatcaa ctcatgtgat 4741 tgaaaacatc taccttccag ctgcgcagac catgaattca ggaactttta acaccacagt 4801 ggatatcaag cttaaacagt ggactgataa acaacttcct aataaagcag tagaggttgc 4861 ttgggagacc ctacaagaag aattttcccg ctttatgaca gaaccgaaag ggaaagagca 4921 tgatgacata tttgataaac ttaaagaggc tgttaaggaa gaaagtatta aacgacacaa 4981 gtggaatgac tttgcggagg acagcttgag ggttattcaa cacaatgctt tggaagaccg 5041 atccatatct gataaacagc aatgggatgc agctatttat tttatggaag aggctctgca 5101 ggctcgtctc aaggatactg aaaatgcaat tgaaaacatg gtgggtccag actggaaaaa 5161 gaggtggtta tactggaaga atcggaccca agaacagtgt gttcacaatg aaaccaagaa 5221 tgaattggag aagatgttga aatgtaatga ggagcaccca gcttatcttg caagtgatga 5281 aataaccaca gtccggaaga accttgaatc ccgaggagta gaagtagatc caagcttgat 5341 taaggatact tggcatcaag tttatagaag acatttttta aaaacagctc taaaccattg 5401 taacctttgt cgaagaggtt tttattacta ccaaaggcat tttgtagatt ctgagttgga 5461 atgcaatgat gtggtcttgt tttggcgtat acagcgcatg cttgctatca ccgcaaatac 5521 tttaaggcaa caacttacaa atactgaagt taggcgatta gagaaaaatg ttaaagaggt 5581 attggaagat tttgctgaag atggtgagaa gaagattaaa ttgcttactg gtaaacgcgt 5641 tcaactggcg gaagacctca agaaagttag agaaattcaa gaaaaacttg atgctttcat 5701 tgaagctctt catcaggaga aaggatccga ctacaaagac catgacggtg attataaaga 5761 tcatgacatc gactacaagg atgacgatga caagtagtct agataattct accgggtagg 5821 ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg 5881 gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc 5941 ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt 6001 cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac 6061 tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag 6121 cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc 6181 cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt cctccggagg 6241 cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc 6301 tccgggcctt tcgacctgca gcccaagctt accatgaccg agtacaagcc cacggtgcgc 6361 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac 6421 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg 6481 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac 6541 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc 6601 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg 6661 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc 6721 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag 6781 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc 6841 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc 6901 acctggtgca tgacccgcaa gcccggtgcc tgaccgcgtc tggaacaatc aacctctgga 6961 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg 7021 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt 7081 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag 7141 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc 7201 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga 7261 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa 7321 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac 7381 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct 7441 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca 7501 gacgagtcgg atctcccttt gggccgcctc cccgcctgga attaattctg cagtcgagac 7561 ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc tgattgtgcc 7621 tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca ggtaccttta 7681 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagagggga 7741 ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg gatctaccac 7801 acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt cagatatcca 7861 ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt agaagaggcc 7921 aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat ggatgacccg 7981 gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca cgtggcccga 8041 gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa gggactttcc 8101 gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg agccctcaga 8161 tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag accagatctg 8221 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc 8281 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct 8341 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat 8401 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggcctt gacattgcta 8461 gcgttttacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg 8521 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta 8581 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg 8641 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga 8701 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 8761 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 8821 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 8881 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 8941 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 9001 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 9061 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 9121 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 9181 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 9241 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 9301 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 9361 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 9421 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 9481 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 9541 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 9601 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 9661 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 9721 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 9781 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 9841 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 9901 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 9961 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 10021 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 10081 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 10141 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 10201 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 10261 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 10321 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 10381 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 10441 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 10501 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 10561 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 10621 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatcaac 10681 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat 10741 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat 10801 catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact tcccacccca 10861 taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg tgattcctct 10921 gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa cttagtagtt 10981 tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagt //