LOCUS V022500 8804 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022500 VERSION V022500 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 2827..4056 /label="LAMP2(NM_013995)" /note="LAMP2(NM_013995)" /gene="LAMP2" CDS 4132..4851 /codon_start=1 /note="mammalian codon-optimized" /product="enhanced blue variant of GFP" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTLTHGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNG IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV LLEFVTAAGITLGMDELYK*" /label="EBFP" misc_feature 4858..5446 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5520..6153 /note="3' long terminal repeat (LTR) from HIV-1" /label="3' LTR" primer_bind complement(6282..6298) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6306..6322 /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(6330..6360) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6375..6396 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6684..7272) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7443..8303) /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(8304..8408) /gene="bla" /label="AmpR promoter" polyA_signal 8456..8590 /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 gccaccatgg tgtgcttccg cctcttcccg gttccgggct cagggctcgt tctggtctgc 2881 ctagtcctgg gagctgtgcg gtcttatgca ttggaactta atttgacaga ttcagaaaat 2941 gccacttgcc tttatgcaaa atggcagatg aatttcacag tacgctatga aactacaaat 3001 aaaacttata aaactgtaac catttcagac catggcactg tgacatataa tggaagcatt 3061 tgtggggatg atcagaatgg tcccaaaata gcagtgcagt tcggacctgg cttttcctgg 3121 attgcgaatt ttaccaaggc agcatctact tattcaattg acagcgtctc attttcctac 3181 aacactggtg ataacacaac atttcctgat gctgaagata aaggaattct tactgttgat 3241 gaacttttgg ccatcagaat tccattgaat gaccttttta gatgcaatag tttatcaact 3301 ttggaaaaga atgatgttgt ccaacactac tgggatgttc ttgtacaagc ttttgtccaa 3361 aatggcacag tgagcacaaa tgagttcctg tgtgataaag acaaaacttc aacagtggca 3421 cccaccatac acaccactgt gccatctcct actacaacac ctactccaaa ggaaaaacca 3481 gaagctggaa cctattcagt taataatggc aatgatactt gtctgctggc taccatgggg 3541 ctgcagctga acatcactca ggataaggtt gcttcagtta ttaacatcaa ccccaataca 3601 actcactcca caggcagctg ccgttctcac actgctctac ttagactcaa tagcagcacc 3661 attaagtatc tagactttgt ctttgctgtg aaaaatgaaa accgatttta tctgaaggaa 3721 gtgaacatca gcatgtattt ggttaatggc tccgttttca gcattgcaaa taacaatctc 3781 agctactggg atgcccccct gggaagttct tatatgtgca acaaagagca gactgtttca 3841 gtgtctggag catttcagat aaataccttt gatctaaggg ttcagccttt caatgtgaca 3901 caaggaaagt attctacagc ccaagagtgt tcgctggatg atgacaccat tctaatccca 3961 attatagttg gtgctggtct ttcaggcttg attatcgtta tagtgattgc ttacgtaatt 4021 ggcagaagaa aaagttatgc tggatatcag actctgtccg gccggactca gatctcgagc 4081 tcaagcttcg aattctgcag tcgacggtac cagagcccag gcccggcagc catggtgagc 4141 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta 4201 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg 4261 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc 4321 accctgaccc acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac 4381 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac 4441 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc 4501 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag 4561 tacaacttca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag 4621 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac 4681 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc 4741 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag 4801 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagtg agtcgacaat 4861 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct 4921 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg 4981 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg 5041 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt 5101 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt 5161 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg 5221 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc 5281 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat 5341 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc 5401 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg tacctttaag 5461 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa agaggggact 5521 ggaagggcta attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac 5581 acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact 5641 gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa 5701 taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga 5761 gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga 5821 gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc 5881 tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc 5941 ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag 6001 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt 6061 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca 6121 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc 6181 agtatttata acttgcaaag aaatgaatat cagagagtga gaggccttga cattgctagc 6241 gttttaccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg 6301 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 6361 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 6421 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 6481 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 6541 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 6601 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 6661 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 6721 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 6781 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 6841 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 6901 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 6961 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 7021 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 7081 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc 7141 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 7201 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 7261 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 7321 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 7381 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 7441 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 7501 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 7561 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 7621 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 7681 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 7741 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 7801 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 7861 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 7921 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 7981 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 8041 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 8101 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 8161 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 8221 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 8281 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 8341 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 8401 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcaactt 8461 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 8521 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 8581 tgtctggatc aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata 8641 ccctattacc actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga 8701 attattttca ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagtttt 8761 taaagaaatt gtatttgtta aatatgtact acaaacttag tagt //