LOCUS V029426 9301 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029426 VERSION V029426 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..3997 /label="Dlk1(NM_010052)" /note="Dlk1(NM_010052)" /gene="Dlk1" CDS 4004..4069 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4089..4588 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4609..5208 /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 5222..5810 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6017..6650 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6779..6795) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6803..6819 /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(6827..6857) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6872..6893 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7181..7769) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7940..8800) /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(8801..8905) /gene="bla" /label="AmpR promoter" polyA_signal 8953..9087 /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 ccatgatcgc gaccggagcc ctcctgcgcg tcctcttgct 2881 cctgctggct ttcggccaca gcacctatgg ggctgaatgc gacccaccct gtgaccccca 2941 gtatggattc tgcgaggctg acaatgtctg caggtgccat gttggctggg agggtcccct 3001 ctgtgacaag tgtgtaactg cccctggctg tgtcaatgga gtctgcaagg aaccatggca 3061 gtgcatctgc aaggatggct gggacgggaa attctgcgaa atagacgttc gggcttgcac 3121 ctcaaccccc tgcgccaaca atggaacttg cgtggacctg gagaaaggcc agtacgaatg 3181 ctcctgcaca cctgggttct ctggaaagga ctgccagcac aaggctgggc cctgcgtgat 3241 caatggttct ccctgccagc acggaggcgc ctgcgtggat gatgagggcc aggcctcgca 3301 tgcttcctgc ctgtgccccc ctggcttctc aggcaacttc tgtgagatcg tagccgcaac 3361 caacagctgt acccctaacc catgcgagaa cgatggcgtc tgcaccgaca tcgggggtga 3421 cttccgttgc cgctgcccag ctggattcgt cgacaagacc tgcagccgcc cggtgagcaa 3481 ctgcgccagt ggcccgtgcc agaacggggg cacctgcctc cagcacaccc aggtgagctt 3541 cgagtgtctg tgcaagcccc cgttcatggg tcccacgtgc gcgaagaagc gcggggctag 3601 ccccgtgcag gtcacccacc tgcccagcgg ctatgggctc acctaccgcc tgacccccgg 3661 ggtgcacgag ctgcctgttc agcagcccga gcaacacatc ctgaaggtgt ccatgaaaga 3721 gctcaacaag agtacccctc tcctcaccga gggacaggcc atctgcttca ccatcctggg 3781 cgtgctcacc agcctggtgg tgctgggcac cgtggccatc gtctttctca acaagtgcga 3841 aacctgggtg tccaacctgc gctacaacca catgcttcgc aagaagaaga acctcctgtt 3901 gcagtataac agcggcgagg agctggcggt caatatcatc ttccccgaga agattgacat 3961 gaccactttc aacaaggagg ctggtgatga ggagatcgga tccgactaca aagaccatga 4021 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt aatctagata 4081 attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg ctttagcagc 4141 cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt ccacatccac 4201 cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt ctactcctcc 4261 cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga 4321 agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt 4381 aggcctttgg ggcagcggcc aatagcagct ttgctccttc gctttctggg ctcagaggct 4441 gggaaggggt gggtccgggg gcgggctcag gggcgggctc aggggcgggg cgggcgcccg 4501 aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt 4561 ctcctcttcc tcatctccgg gcctttcgac ctgcagccca agcttaccat gaccgagtac 4621 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc 4681 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag 4741 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg 4801 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg 4861 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc 4921 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc 4981 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg 5041 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg 5101 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg 5161 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacc gcgtctggaa 5221 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 5281 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 5341 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 5401 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 5461 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 5521 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 5581 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct 5641 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 5701 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 5761 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattaa 5821 ttctgcagtc gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc 5881 aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca 5941 cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta 6001 aaagaaaaga ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat 6061 ctgtggatct accacacaca aggctacttc cctgattagc agaactacac accagggcca 6121 ggggtcagat atccactgac ctttggatgg tgctacaagc tagtaccagt tgagccagat 6181 aaggtagaag aggccaataa aggagagaac accagcttgt tacaccctgt gagcctgcat 6241 gggatggatg acccggagag agaagtgtta gagtggaggt ttgacagccg cctagcattt 6301 catcacgtgg cccgagagct gcatccggag tacttcaaga actgctgata tcgagcttgc 6361 tacaagggac tttccgctgg ggactttcca gggaggcgtg gcctgggcgg gactggggag 6421 tggcgagccc tcagatcctg catataagca gctgcttttt gcctgtactg ggtctctctg 6481 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc 6541 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg 6601 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc 6661 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag 6721 gccttgacat tgctagcgtt ttaccgtcga cctctagcta gagcttggcg taatcatggt 6781 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 6841 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 6901 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 6961 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 7021 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 7081 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 7141 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 7201 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 7261 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 7321 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 7381 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 7441 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 7501 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 7561 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 7621 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 7681 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 7741 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 7801 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 7861 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 7921 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 7981 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 8041 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 8101 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 8161 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 8221 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 8281 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 8341 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 8401 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 8461 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 8521 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 8581 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 8641 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 8701 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 8761 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 8821 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 8881 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg 8941 gatcgggaga tcaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 9001 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 9061 atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca aaatcatccc 9121 aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca tttactctaa 9181 acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact 9241 acaaacttag tagtttttaa agaaattgta tttgttaaat atgtactaca aacttagtag 9301 t //