LOCUS V021415 8658 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V021415 VERSION V021415 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 2815..3309 /label="ISG15(NM_005101)" /note="ISG15(NM_005101)" /gene="ISG15" misc_feature 3329..3901 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /label="IRES" CDS 3903..4613 /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" misc_feature 4627..5215 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5422..6055 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6183..6199) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6207..6223 /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(6231..6261) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6276..6297 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6585..7173) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7344..8204) /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(8205..8309) /gene="bla" /label="AmpR promoter" polyA_signal 8357..8491 /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 tctatttccg gtgaattcgc caccatgggc 2821 tgggacctga cggtgaagat gctggcgggc aacgaattcc aggtgtccct gagcagctcc 2881 atgtcggtgt cagagctgaa ggcgcagatc acccagaaga tcggcgtgca cgccttccag 2941 cagcgtctgg ctgtccaccc gagcggtgtg gcgctgcagg acagggtccc ccttgccagc 3001 cagggcctgg gccccggcag cacggtcctg ctggtggtgg acaaatgcga cgaacctctg 3061 agcatcctgg tgaggaataa caagggccgc agcagcacct acgaggtacg gctgacgcag 3121 accgtggccc acctgaagca gcaagtgagc gggctggagg gtgtgcagga cgacctgttc 3181 tggctgacct tcgaggggaa gcccctggag gaccagctcc cgctggggga gtacggcctc 3241 aagcccctga gcaccgtgtt catgaatctg cgcctgcggg gaggcggcac agagcctggc 3301 gggcggagct aagcggccgc ggatcccgcc cctctccctc ccccccccct aacgttactg 3361 gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat 3421 tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc 3481 ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag 3541 cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc 3601 ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac 3661 ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca 3721 aatggctcac ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt 3781 gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa 3841 aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata 3901 atatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca 3961 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg 4021 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc 4081 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc 4141 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc 4201 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg 4261 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg 4321 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg 4381 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg 4441 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg 4501 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg 4561 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag tgaacgcgtc 4621 tggaacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat 4681 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct 4741 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag 4801 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc 4861 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc 4921 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct 4981 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg 5041 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg 5101 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg 5161 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga 5221 attaattctg cagtcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag 5281 ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag 5341 tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact 5401 ttttaaaaga aaagagggga ctggaagggc taattcactc ccaacgaaga caagatatcc 5461 ttgatctgtg gatctaccac acacaaggct acttccctga ttagcagaac tacacaccag 5521 ggccaggggt cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc 5581 cagataaggt agaagaggcc aataaaggag agaacaccag cttgttacac cctgtgagcc 5641 tgcatgggat ggatgacccg gagagagaag tgttagagtg gaggtttgac agccgcctag 5701 catttcatca cgtggcccga gagctgcatc cggagtactt caagaactgc tgatatcgag 5761 cttgctacaa gggactttcc gctggggact ttccagggag gcgtggcctg ggcgggactg 5821 gggagtggcg agccctcaga tcctgcatat aagcagctgc tttttgcctg tactgggtct 5881 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 5941 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 6001 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt 6061 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt 6121 gagaggcctt gacattgcta gcgtttaccg tcgacctcta gctagagctt ggcgtaatca 6181 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga 6241 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt 6301 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga 6361 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc 6421 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg 6481 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc 6541 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 6601 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 6661 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 6721 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat 6781 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 6841 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 6901 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 6961 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 7021 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 7081 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag 7141 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg 7201 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa 7261 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata 7321 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg 7381 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata 7441 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg 7501 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct 7561 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt 7621 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc 7681 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga 7741 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt 7801 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc 7861 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa 7921 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca 7981 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca 8041 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct 8101 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc 8161 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa 8221 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt 8281 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc 8341 gacggatcgg gagatcaact tgtttattgc agcttataat ggttacaaat aaagcaatag 8401 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 8461 actcatcaat gtatcttatc atgtctggat caactggata actcaagcta accaaaatca 8521 tcccaaactt cccaccccat accctattac cactgccaat tacctgtggt ttcatttact 8581 ctaaacctgt gattcctctg aattattttc attttaaaga aattgtattt gttaaatatg 8641 tactacaaac ttagtagt //