LOCUS V018806 10206 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V018806 VERSION V018806 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 2807..3514 /codon_start=1 /note="mammalian codon-optimized" /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /transl_table=1 /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" /label="mCherry" CDS 3551..4933 /label="Coro1a(NM_001301374)" /note="Coro1a(NM_001301374)" /gene="Coro1a" CDS 4934..4999 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 5041..5540 /note="mouse phosphoglycerate kinase 1 promoter" /label="PGK promoter" CDS 5561..6160 /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 6174..6762 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6969..7602 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(7731..7747) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7755..7771 /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(7779..7809) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7824..7845 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8133..8721) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8892..9752) /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(9753..9857) /gene="bla" /label="AmpR promoter" polyA_signal 9905..10039 /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 tctaccggtc gccaccatgg tgagcaaggg 2821 cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg 2881 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg 2941 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat 3001 cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc 3061 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga 3121 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta 3181 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac 3241 catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga 3301 gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac 3361 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga 3421 catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg 3481 ccactccacc ggcggcatgg acgagctgta caagtccgga ctcagatctc gagctcaagc 3541 ttcgaattcg atgagccggc aggtggttcg ctccagcaaa ttccgccacg tgtttggaca 3601 gccagccaag gctgaccagt gctatgagga tgtgcgcgtc tcacaaacca cttgggacag 3661 tggcttctgc gctgtcaacc ccaagttcat ggctctgatc tgtgaggcca gtgggggagg 3721 ggccttcctg gtgctacccc taggcaagac tggacgagta gacaagaacg tgcccctggt 3781 ctgcggccac actgcccctg tgctagacat cgcctggtgt ccacacaatg acaatgtcat 3841 tgccagtggc tctgaggact gcacagttat ggtgtgggag atcccggatg ggggcctggt 3901 gctgcccttg cgggagcctg tcatcacctt ggagggccac accaagaggg ttggcatcgt 3961 ggcctggcac cccacagccc agaatgtgct tctcagcgca ggttgtgaca atgtgatcct 4021 ggtgtgggac gtgggcactg gggcagctgt gctgacgctg ggcccggatg tgcacccgga 4081 cacaatctac agcgtggact ggagccggga cggtgctctc atctgcacct cctgccgtga 4141 caagcgagtt cgtgtcatcg agcctcgcaa aggcactgtt gtggctgaaa aggaccgtcc 4201 ccatgagggg actcggccag tgcacgctgt gtttgtgtca gaagggaaga ttctaaccac 4261 gggtttcagt cgcatgagtg agcggcaggt ggcgctgtgg gacacaaagc acctggagga 4321 gccactgtcc ctgcaggagc tggacacaag cagcggagtc ctgctacctt tttttgatcc 4381 tgataccaac attgtctacc tctgtggcaa gggtgacagc tctatccggt attttgagat 4441 cacttctgag gctccgttcc tgcattatct ttccatgttc agttccaagg agtctcagcg 4501 tggtatgggc tacatgccca aacgtggcct ggaagtgaat aagtgtgaga ttgccagatt 4561 ctataagctt catgagcgga agtgtgagcc cattgccatg acagtgccta gaaagtcgga 4621 cctgttccag gaggacctgt atccgcccac tgcaggaccc gaccctgccc tcacagctga 4681 ggaatggctg ggtggtcggg atgctggccc cctcctcatt tccctcaagg atggctacgt 4741 gcccccaaag agcagggagc tgagggtcaa caggggcctg gacagcgctc gcagaagagc 4801 tacaccagag cccagcggca cgcccagctc ggataccgtg tcaaggctgg aggaggacgt 4861 gaggaatctg aacgccatag tgcaaaagct acaagagcgc ctggacaggc tggaggagac 4921 tgttcaggcc aaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta 4981 caaggatgac gatgacaagt gaggatccac cggatctaga taactgatca taattctacc 5041 gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg 5101 gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc 5161 gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca 5221 ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac 5281 gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt 5341 ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg 5401 gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct 5461 ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt 5521 cctcatctcc gggcctttcg acctgcagcc caagcttacc atgaccgagt acaagcccac 5581 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt 5641 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac 5701 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc 5761 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt 5821 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca 5881 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac 5941 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg 6001 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa 6061 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg 6121 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga ccgcgtctgg aacaatcaac 6181 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta 6241 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt 6301 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg 6361 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg 6421 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca 6481 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca 6541 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg 6601 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag 6661 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc 6721 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaatt aattctgcag 6781 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga 6841 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt 6901 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa 6961 gaggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat 7021 ctaccacaca caaggctact tccctgatta gcagaactac acaccagggc caggggtcag 7081 atatccactg acctttggat ggtgctacaa gctagtacca gttgagccag ataaggtaga 7141 agaggccaat aaaggagaga acaccagctt gttacaccct gtgagcctgc atgggatgga 7201 tgacccggag agagaagtgt tagagtggag gtttgacagc cgcctagcat ttcatcacgt 7261 ggcccgagag ctgcatccgg agtacttcaa gaactgctga tatcgagctt gctacaaggg 7321 actttccgct ggggactttc cagggaggcg tggcctgggc gggactgggg agtggcgagc 7381 cctcagatcc tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc 7441 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa 7501 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga 7561 gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc 7621 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggccttgac 7681 attgctagcg ttttaccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg 7741 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata 7801 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 7861 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 7921 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 7981 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 8041 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 8101 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 8161 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 8221 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 8281 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 8341 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 8401 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 8461 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 8521 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta 8581 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 8641 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 8701 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 8761 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 8821 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 8881 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 8941 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 9001 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta 9061 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 9121 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 9181 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt 9241 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 9301 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 9361 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 9421 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 9481 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact 9541 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg 9601 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 9661 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 9721 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 9781 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 9841 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga 9901 gatcaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 9961 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 10021 atcttatcat gtctggatca actggataac tcaagctaac caaaatcatc ccaaacttcc 10081 caccccatac cctattacca ctgccaatta cctgtggttt catttactct aaacctgtga 10141 ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt 10201 agtagt //