LOCUS V033356 9661 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V033356 VERSION V033356 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..4357 /label="Pdia3(NM_007952)" /note="Pdia3(NM_007952)" /gene="Pdia3" CDS 4364..4429 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4449..4948 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4969..5568 /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 5582..6170 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6377..7010 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7139..7155) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7163..7179 /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(7187..7217) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7232..7253 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7541..8129) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8300..9160) /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(9161..9265) /gene="bla" /label="AmpR promoter" polyA_signal 9313..9447 /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 ccatgcgctt cagctgccta gctctgctcc cgggcgtggc 2881 gctgctgctc gcctcggccc gtctcgccgc cgcctccgat gtgttggaac tgacggacga 2941 aaacttcgag agtcgcgtct ccgacacggg ctcggcgggg ctcatgctag tcgagttctt 3001 cgccccctgg tgtggacatt gcaagaggct tgcccctgag tatgaagctg cagcaaccag 3061 attaaaagga atagtcccat tagcaaaggt ggattgcact gccaacacaa acacctgtaa 3121 taagtatggg gtcagtggct acccaactct taagatcttt agagatggtg aagaagcggg 3181 tgcttatgat gggcctagga ctgctgatgg aattgtcagc cacttgaaga aacaagcagg 3241 accagcttca gttcctctca ggactgagga agaatttaag aagttcatta gtgataaaga 3301 tgcctcagtg gtgggttttt tcagggattt attcagtgat gggcactctg aattcctaaa 3361 agcagccagc aacttgagag ataactaccg atttgcacac accaacattg agtctctggt 3421 gaaggagtac gatgataatg gagaggggat cactatattc cgtccattac atcttgctaa 3481 caagtttgaa gacaaaactg tggcatatac tgaaaagaaa atgaccagtg gcaagatcaa 3541 gaaatttatt caggatagca tttttggtct ctgtcctcat atgacggaag ataataaaga 3601 tttgatacaa ggcaaggact tactcaccgc ttactatgat gtggactatg aaaagaatgc 3661 taaaggttct aactactgga gaaacagggt catgatggtg gcaaagaaat tccttgatgc 3721 tggacacaaa ctcaactttg ctgtagctag ccgtaaaacc tttagccatg aactgtcaga 3781 ttttggctta gaaagcacta ctggagaggt tcctgttgtg gctatcagaa ctgctaaagg 3841 agagaagttt gtcatgcagg aggagttctc gcgagatggc aaggctcttg aacagttcct 3901 gcaagaatac tttgatggca acttgaagag atacctgaag tctgaaccca tcccagagtc 3961 caacgaaggg cctgtcaagg ttgtggtagc agagaatttt gatgacatag tgaatgaaga 4021 agataaggac gtgctgattg aattttacgc cccttggtgt ggccactgta agaatctgga 4081 acccaagtat aaagagctgg gagaaaaact cagcaaagat ccaaatattg tcatagccaa 4141 gatggatgcc acagccaatg atgtgccttc tccatatgaa gtcaagggtt ttcctaccat 4201 ctacttctca ccagccaaca agaagctaac tccaaagaag tatgaaggtg gccgtgaatt 4261 aaatgatttt attagctatc tacaacgaga agctacaaac ccccctataa ttcaagaaga 4321 aaaacctaag aagaagaaga aggcacaaga ggacctcgga tccgactaca aagaccatga 4381 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt agtctagata 4441 attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg ctttagcagc 4501 cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt ccacatccac 4561 cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt ctactcctcc 4621 cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga 4681 agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt 4741 aggcctttgg ggcagcggcc aatagcagct ttgctccttc gctttctggg ctcagaggct 4801 gggaaggggt gggtccgggg gcgggctcag gggcgggctc aggggcgggg cgggcgcccg 4861 aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt 4921 ctcctcttcc tcatctccgg gcctttcgac ctgcagccca agcttaccat gaccgagtac 4981 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc 5041 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag 5101 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg 5161 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg 5221 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc 5281 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc 5341 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg 5401 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg 5461 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg 5521 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacc gcgtctggaa 5581 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 5641 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 5701 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 5761 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 5821 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 5881 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 5941 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct 6001 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 6061 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 6121 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattaa 6181 ttctgcagtc gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc 6241 aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca 6301 cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta 6361 aaagaaaaga ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat 6421 ctgtggatct accacacaca aggctacttc cctgattagc agaactacac accagggcca 6481 ggggtcagat atccactgac ctttggatgg tgctacaagc tagtaccagt tgagccagat 6541 aaggtagaag aggccaataa aggagagaac accagcttgt tacaccctgt gagcctgcat 6601 gggatggatg acccggagag agaagtgtta gagtggaggt ttgacagccg cctagcattt 6661 catcacgtgg cccgagagct gcatccggag tacttcaaga actgctgata tcgagcttgc 6721 tacaagggac tttccgctgg ggactttcca gggaggcgtg gcctgggcgg gactggggag 6781 tggcgagccc tcagatcctg catataagca gctgcttttt gcctgtactg ggtctctctg 6841 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc 6901 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg 6961 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc 7021 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag 7081 gccttgacat tgctagcgtt ttaccgtcga cctctagcta gagcttggcg taatcatggt 7141 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 7201 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 7261 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 7321 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 7381 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 7441 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 7501 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 7561 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 7621 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 7681 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 7741 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 7801 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 7861 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 7921 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 7981 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 8041 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 8101 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 8161 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 8221 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 8281 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 8341 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 8401 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 8461 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 8521 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 8581 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 8641 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 8701 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 8761 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 8821 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 8881 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 8941 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 9001 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 9061 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 9121 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 9181 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 9241 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg 9301 gatcgggaga tcaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 9361 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 9421 atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca aaatcatccc 9481 aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca tttactctaa 9541 acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact 9601 acaaacttag tagtttttaa agaaattgta tttgttaaat atgtactaca aacttagtag 9661 t //