LOCUS V022627 8891 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022627 VERSION V022627 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 2826..3617 /label="SNAI1(NM_005985)" /note="SNAI1(NM_005985)" /gene="SNAI1" CDS 3624..3647 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" promoter 3679..4178 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 4199..4798 /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 4812..5400 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5607..6240 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6369..6385) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6393..6409 /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(6417..6447) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6462..6483 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6771..7359) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7530..8390) /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(8391..8495) /gene="bla" /label="AmpR promoter" polyA_signal 8543..8677 /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 ccaccatgcc gcgctctttc ctcgtcagga agccctccga ccccaatcgg aagcctaact 2881 acagcgagct gcaggactct aatccagagt ttaccttcca gcagccctac gaccaggccc 2941 acctgctggc agccatccca cctccggaga tcctcaaccc caccgcctcg ctgccaatgc 3001 tcatctggga ctctgtcctg gcgccccaag cccagccaat tgcctgggcc tcccttcggc 3061 tccaggagag tcccagggtg gcagagctga cctccctgtc agatgaggac agtgggaaag 3121 gctcccagcc ccccagccca ccctcaccgg ctccttcgtc cttctcctct acttcagtct 3181 cttccttgga ggccgaggcc tatgctgcct tcccaggctt gggccaagtg cccaagcagc 3241 tggcccagct ctctgaggcc aaggatctcc aggctcgaaa ggccttcaac tgcaaatact 3301 gcaacaagga atacctcagc ctgggtgccc tcaagatgca catccgaagc cacacgctgc 3361 cctgcgtctg cggaacctgc gggaaggcct tctctaggcc ctggctgcta caaggccatg 3421 tccggaccca cactggcgag aagcccttct cctgtcccca ctgcagccgt gccttcgctg 3481 accgctccaa cctgcgggcc cacctccaga cccactcaga tgtcaagaag taccagtgcc 3541 aggcgtgtgc tcggaccttc tcccgaatgt ccctgctcca caagcaccaa gagtccggct 3601 gctcaggatg tccccgcgaa ttcgattaca aggatgacga cgataagtga ggatcccgcg 3661 actctagata attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg 3721 ctttagcagc cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt 3781 ccacatccac cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt 3841 ctactcctcc cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt 3901 gacaaatgga agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat 3961 ggaagcgggt aggcctttgg ggcagcggcc aatagcagct ttgctccttc gctttctggg 4021 ctcagaggct gggaaggggt gggtccgggg gcgggctcag gggcgggctc aggggcgggg 4081 cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg 4141 ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac ctgcagccca agcttaccat 4201 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg 4261 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg 4321 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat 4381 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag 4441 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc 4501 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc 4561 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag 4621 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga 4681 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga 4741 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacc 4801 gcgtctggaa caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta 4861 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta 4921 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt 4981 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg 5041 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt 5101 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag 5161 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc 5221 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc 5281 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc 5341 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc 5401 ctggaattaa ttctgcagtc gagacctaga aaaacatgga gcaatcacaa gtagcaatac 5461 agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt 5521 tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag 5581 ccacttttta aaagaaaaga ggggactgga agggctaatt cactcccaac gaagacaaga 5641 tatccttgat ctgtggatct accacacaca aggctacttc cctgattagc agaactacac 5701 accagggcca ggggtcagat atccactgac ctttggatgg tgctacaagc tagtaccagt 5761 tgagccagat aaggtagaag aggccaataa aggagagaac accagcttgt tacaccctgt 5821 gagcctgcat gggatggatg acccggagag agaagtgtta gagtggaggt ttgacagccg 5881 cctagcattt catcacgtgg cccgagagct gcatccggag tacttcaaga actgctgata 5941 tcgagcttgc tacaagggac tttccgctgg ggactttcca gggaggcgtg gcctgggcgg 6001 gactggggag tggcgagccc tcagatcctg catataagca gctgcttttt gcctgtactg 6061 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac 6121 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt 6181 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca 6241 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag 6301 agagtgagag gccttgacat tgctagcgtt ttaccgtcga cctctagcta gagcttggcg 6361 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac 6421 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca 6481 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat 6541 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc 6601 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 6661 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 6721 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg 6781 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 6841 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 6901 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 6961 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 7021 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 7081 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt 7141 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc 7201 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa 7261 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt 7321 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 7381 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 7441 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 7501 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 7561 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 7621 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc 7681 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 7741 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 7801 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg 7861 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt 7921 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc 7981 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt 8041 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc 8101 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc 8161 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa 8221 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac 8281 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa 8341 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt 8401 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa 8461 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct 8521 gacgtcgacg gatcgggaga tcaacttgtt tattgcagct tataatggtt acaaataaag 8581 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt 8641 gtccaaactc atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca 8701 aaatcatccc aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca 8761 tttactctaa acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta 8821 aatatgtact acaaacttag tagtttttaa agaaattgta tttgttaaat atgtactaca 8881 aacttagtag t //