LOCUS V032764 9415 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032764 VERSION V032764 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..4090 /label="Pip4k2b(NM_054051)" /note="Pip4k2b(NM_054051)" /gene="Pip4k2b" CDS 4097..4123 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 4127..4153 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 4157..4183 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" promoter 4203..4702 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4723..5322 /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 5336..5924 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6131..6764 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6893..6909) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6917..6933 /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(6941..6971) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6986..7007 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7295..7883) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8054..8914) /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(8915..9019) /gene="bla" /label="AmpR promoter" polyA_signal 9067..9201 /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 ccatgtcgtc caactgcacc agcaccacgg cggtggcggt 2881 ggcgccgctc agcgccagca aaaccaagac caagaagaag catttcgtgt gccagaaagt 2941 gaagctattc cgggccagcg agccgattct cagcgtcctg atgtgggggg tgaaccacac 3001 gatcaatgag ctgagcaatg tccctgttcc tgtcatgcta atgccggatg acttcaaagc 3061 ttacagcaag atcaaggtgg acaatcatct cttcaataag gaaaacctcc ccagccgctt 3121 caagttcaag gagtactgcc ccatggtgtt ccgaaacctc agggagaggt tcggcatcga 3181 tgaccaggac taccagaatt cagtgacacg cagtgcacct atcaacagtg acagccaggg 3241 caggtgtggc actcgcttcc tcaccaccta tgaccggcgt ttcgtcatca agactgtgtc 3301 aagtgaggat gtggccgaga tgcacaacat cctaaagaaa taccaccagt tcatagtgga 3361 atgtcacggc aataccctct tgccacagtt cctgggcatg taccgcctga ccgtggatgg 3421 ggtggaaacc tacatggtgg tcaccaggaa tgtattcagt catcggctta ctgtgcatcg 3481 caaatacgac ctgaagggtt ccactgttgc cagagaagcc agtgacaagg aaaaggctaa 3541 ggacttgcca actttcaaag ataatgactt cctcaacgaa gggcagaagc tgcacgtggg 3601 agaagagagc aaaaagaact tcctggaaaa actgaagcgg gacgttgagt tcctggccca 3661 gctgaagatc atggattaca gtctcctggt gggcatccac gacgtggacc gggcagagca 3721 ggaggagatg gaggtggaag agcgagcgga ggaggaggaa tgcgagaatg atggggtggg 3781 tggcagcctg ctctgctcct atggcacgcc tccagacagc cctggcaacc tcctgagctt 3841 cccccggttc ttcggcccag gagagttcga cccctctgtg gacgtctatg ccatgaaaag 3901 ccatgaaagt gcccctaaga aggaggtcta tttcatggcc atcattgaca tcctcacacc 3961 ctatgacgct aagaagaaag ctgcacacgc cgccaaaacg gtgaaacatg gggcaggggc 4021 tgagatctcc actgtgaacc cggagcagta ctcaaaacgc ttcaacgagt tcatgtccaa 4081 catcctgacg ggatcctacc catacgacgt acctgattat gcaacttatc cttatgatgt 4141 acctgattat gctagctatc cttatgatgt cccagactac gcatagtcta gataattcta 4201 ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct 4261 gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag 4321 gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt 4381 caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc 4441 acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct 4501 ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag 4561 gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc 4621 ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc 4681 ttcctcatct ccgggccttt cgacctgcag cccaagctta ccatgaccga gtacaagccc 4741 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg 4801 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc 4861 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc 4921 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg 4981 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag 5041 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc 5101 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc 5161 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc 5221 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa 5281 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaccgcgtct ggaacaatca 5341 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt 5401 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc 5461 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc 5521 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg 5581 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc 5641 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg 5701 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg 5761 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc 5821 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct 5881 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttaattctgc 5941 agtcgagacc tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct 6001 gattgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag 6061 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa 6121 aagaggggac tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg 6181 atctaccaca cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc 6241 agatatccac tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta 6301 gaagaggcca ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg 6361 gatgacccgg agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac 6421 gtggcccgag agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag 6481 ggactttccg ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga 6541 gccctcagat cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga 6601 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata 6661 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta 6721 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca 6781 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggccttg 6841 acattgctag cgttttaccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc 6901 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 6961 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 7021 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 7081 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 7141 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 7201 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 7261 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 7321 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 7381 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 7441 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 7501 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 7561 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 7621 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 7681 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 7741 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 7801 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 7861 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 7921 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 7981 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 8041 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat 8101 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct 8161 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt 8221 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat 8281 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta 8341 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg 8401 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt 8461 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg 8521 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg 8581 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc 8641 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa 8701 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac 8761 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt 8821 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg 8881 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa 8941 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata 9001 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg 9061 gagatcaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 9121 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 9181 gtatcttatc atgtctggat caactggata actcaagcta accaaaatca tcccaaactt 9241 cccaccccat accctattac cactgccaat tacctgtggt ttcatttact ctaaacctgt 9301 gattcctctg aattattttc attttaaaga aattgtattt gttaaatatg tactacaaac 9361 ttagtagttt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt //