LOCUS V025599 9765 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025599 VERSION V025599 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 2830..2856 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 2863..4521 /label="ATP5F1A(NM_004046)" /note="ATP5F1A(NM_004046)" /gene="ATP5F1A" promoter 4553..5052 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5073..5672 /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 5686..6274 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6481..7114 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7243..7259) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7267..7283 /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(7291..7321) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7336..7357 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7645..8233) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8404..9264) /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(9265..9369) /gene="bla" /label="AmpR promoter" polyA_signal 9417..9551 /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 gccaccatgt acccatacga cgtaccagat tacgctgaat tcatgctgtc cgtgcgcgtt 2881 gctgcggccg tggtccgcgc ccttcctcgg cgggccggac tggtctccag aaatgctttg 2941 ggttcatctt tcattgctgc aaggaacttc catgcctcta acactcatct tcaaaagact 3001 gggactgctg agatgtcctc tattcttgaa gagcgtattc ttggagctga tacctctgtt 3061 gatcttgaag aaactgggcg tgtcttaagt attggtgatg gtattgcccg cgtacatggg 3121 ctgaggaatg ttcaagcaga agaaatggta gagttttctt caggcttaaa gggtatgtcc 3181 ttgaacttgg aacctgacaa tgttggtgtt gtcgtgtttg gaaatgataa actaattaag 3241 gaaggagata tagtgaagag gacaggagcc attgtggacg ttccagttgg tgaggagctg 3301 ttgggtcgtg tagttgatgc ccttggtaat gctattgatg gaaagggtcc aattggttcc 3361 aagacgcgta ggcgagttgg tctgaaagcc cccggtatca ttcctcgaat ttcagtgcgg 3421 gaaccaatgc agactggcat taaggctgtg gatagcttgg tgccaattgg tcgtggtcag 3481 cgtgaactga ttattggtga ccgacagact gggaaaacct caattgctat tgacacaatc 3541 attaaccaga aacgtttcaa tgatggatct gatgaaaaga agaagctgta ctgtatttat 3601 gttgctattg gtcaaaagag atccactgtt gcccagttgg tgaagagact tacagatgca 3661 gatgccatga agtacaccat tgtggtgtcg gctacggcct cggatgctgc cccacttcag 3721 tacctggctc cttactctgg ctgttccatg ggagagtatt ttagagacaa tggcaaacat 3781 gctttgatca tctatgacga cttatccaaa caggctgttg cttaccgtca gatgtctctg 3841 ttgctccgcc gaccccctgg tcgtgaggcc tatcctggtg atgtgttcta cctacactcc 3901 cggttgctgg agagagcagc caaaatgaac gatgcttttg gtggtggctc cttgactgct 3961 ttgccagtca tagaaacaca ggctggtgat gtgtctgctt acattccaac aaatgtcatt 4021 tccatcactg acggacagat cttcttggaa acagaattgt tctacaaagg tatccgccct 4081 gcaattaacg ttggtctgtc tgtatctcgt gtcggatccg ctgcccaaac cagggctatg 4141 aagcaggtag caggtaccat gaagctggaa ttggctcagt atcgtgaggt tgctgctttt 4201 gcccagttcg gttctgacct cgatgctgcc actcaacaac ttttgagtcg tggcgtgcgt 4261 ctaactgagt tgctgaagca aggacagtat tctcccatgg ctattgaaga acaagtggct 4321 gttatctatg cgggtgtaag gggatatctt gataaactgg agcccagcaa gattacaaag 4381 tttgagaatg ctttcttgtc tcatgtcgtc agccagcacc aagccttgtt gggcactatc 4441 agggctgatg gaaagatctc agaacaatca gatgcaaagc tgaaagagat tgtaacaaat 4501 ttcttggctg gatttgaagc ttaaggatcc cgcgactcta gataattcta ccgggtaggg 4561 gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg 4621 cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg 4681 gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc 4741 ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact 4801 agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc 4861 ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc 4921 gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc 4981 ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct 5041 ccgggccttt cgacctgcag cccaagctta ccatgaccga gtacaagccc acggtgcgcc 5101 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact 5161 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc 5221 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg 5281 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg 5341 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg 5401 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg 5461 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg 5521 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct 5581 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca 5641 cctggtgcat gacccgcaag cccggtgcct gaccgcgtct ggaacaatca acctctggat 5701 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 5761 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 5821 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 5881 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 5941 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 6001 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 6061 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc 6121 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 6181 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 6241 acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttaattctgc agtcgagacc 6301 tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct 6361 ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa 6421 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aagaggggac 6481 tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca 6541 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 6601 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 6661 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 6721 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 6781 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 6841 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 6901 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 6961 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 7021 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 7081 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt 7141 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggccttg acattgctag 7201 cgttttaccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt 7261 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa 7321 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc 7381 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag 7441 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 7501 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 7561 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 7621 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 7681 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 7741 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 7801 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 7861 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 7921 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 7981 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 8041 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat 8101 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 8161 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa 8221 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga 8281 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct 8341 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga 8401 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc 8461 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg 8521 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat 8581 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat 8641 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg 8701 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc 8761 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa 8821 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc 8881 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt 8941 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag 9001 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt 9061 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag 9121 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac 9181 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc 9241 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca 9301 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg 9361 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatcaact 9421 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata 9481 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc 9541 atgtctggat caactggata actcaagcta accaaaatca tcccaaactt cccaccccat 9601 accctattac cactgccaat tacctgtggt ttcatttact ctaaacctgt gattcctctg 9661 aattattttc attttaaaga aattgtattt gttaaatatg tactacaaac ttagtagttt 9721 ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt //