LOCUS V035814 10205 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035814 VERSION V035814 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers LTR 234..414 /note="truncated 5' long terminal repeat (LTR) from HIV-1" /label="5' LTR (truncated)" misc_feature 458..583 /note="packaging signal of human immunodeficiency virus type 1" /label="HIV-1 Ψ" misc_feature 1076..1309 /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." /label="RRE" CDS 1493..1537 /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 1804..1920 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2010..2213 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2302..4308 /label="GLS(NM_014905)" /note="GLS(NM_014905)" /gene="GLS" CDS 4315..4341 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 4345..4371 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 4375..4401 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 4408..5127 /codon_start=1 /note="mammalian codon-optimized" /product="the original enhanced GFP (Yang et al., 1996)" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV LLEFVTAAGITLGMDELYK*" /label="EGFP" promoter 5175..5386 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 5399..5667 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 5694..6293 /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 6300..6888 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6962..7195 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7267..7388 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7407..7542 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7580..7596) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7604..7620 /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(7628..7658) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7673..7694 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7982..8570) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8741..9601) /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(9602..9706) /gene="bla" /label="AmpR promoter" primer_bind 10180..10196 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" ORIGIN 1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca 61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta 121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga 181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc 241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg 421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct 481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt 541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag 601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt 661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt 721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg 781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag 841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag 901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg 961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag 1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag 1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc 1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga 1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc 1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg 1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata 1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa 1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga 1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa 1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat 1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt 1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg 1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt 1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat 1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt 1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag 2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct 2281 agagctagcg aattcgccac catgatgcgg ctgcgaggct cggggatgct gcgggacctg 2341 ctcctgcggt cgcccgccgg cgtgagcgcg actctgcggc gggcacagcc cttggtcacc 2401 ctgtgccggc gtccccgagg cgggggacgg ccggccgcgg gcccggctgc cgccgcgcga 2461 ctccacccgt ggtggggcgg gggcggctgg ccggcggagc ccctcgcgcg gggcctgtcc 2521 agctctcctt cggagatctt gcaggagctg ggcaagggga gcacgcatcc gcagcccggg 2581 gtgtcgccac ccgctgcccc ggcggcgccc ggccccaagg acggccccgg ggagacggac 2641 gcgtttggca acagcgaggg caaagagctg gtggcctcag gtgaaaataa aataaaacag 2701 ggtctgttac ctagcttgga agatttgctg ttctatacaa ttgctgaagg acaagagaaa 2761 atacctgttc ataaatttat tacagcactc aaatctacag gattgcgaac gtctgatccc 2821 aggttgaaag agtgtatgga tatgttaaga ttaactcttc aaacaacatc agatggtgtc 2881 atgctagaca aagatctttt taaaaaatgt gttcagagca acattgtttt gttgacacaa 2941 gcatttagaa gaaagtttgt gattcctgac tttatgtctt ttacctcaca cattgatgag 3001 ttatatgaaa gtgctaaaaa gcagtctgga ggaaaggttg cagattatat tcctcaactg 3061 gccaaattca gtcccgattt gtggggtgtg tctgtttgta cagtagatgg acagaggcat 3121 tctactggag ataccaaagt tcccttctgt cttcagtcct gtgtaaaacc tttgaaatat 3181 gccattgctg ttaatgatct tggaactgaa tatgtgcatc gatatgttgg aaaagagccg 3241 agtggactaa gattcaacaa actatttttg aatgaagatg ataaaccaca taatcctatg 3301 gtaaatgctg gagcaattgt tgtgacttca ctaataaagc aaggagtaaa taatgctgaa 3361 aaatttgact atgtcatgca gtttttgaat aagatggctg gtaatgaata tgttggattc 3421 agtaatgcaa cgtttcagtc tgaaagagaa agtggagatc gaaattttgc aataggatat 3481 tacttaaaag aaaagaagtg ttttccagaa ggcacagaca tggttggtat attagacttc 3541 tacttccagc tgtgctccat tgaagtgact tgtgaatcag ccagtgtgat ggctgcgaca 3601 ctggctaatg gtggtttctg cccaattact ggtgaaagag tactgagccc tgaagcagtt 3661 cgaaatacat tgagtttgat gcattcctgt ggcatgtatg acttctcagg gcagtttgct 3721 ttccatgttg gtcttcctgc aaaatctgga gttgctgggg gcattctttt agttgtcccc 3781 aatgttatgg gtatgatgtg ctggtctcct cctctggata agatgggcaa cagtgttaag 3841 ggaattcact tttgtcacga tcttgtttct ctgtgtaatt tccataacta tgataatttg 3901 agacactttg caaaaaaact tgatcctcga agagaaggtg gtgatcaaag ggtaaagtca 3961 gtgataaatc ttttgtttgc tgcatatact ggagatgtgt ctgcacttcg aagatttgct 4021 ttgtcagcta tggacatgga acagcgggac tatgattcta gaacagcact ccatgtagct 4081 gctgcagagg gtcatgttga agttgttaaa tttttgctgg aagcctgcaa agtaaaccct 4141 ttccccaagg acaggtggaa taacactccc atggatgaag cactgcactt tggacaccat 4201 gatgtattta aaattctcca agaataccaa gtccagtaca cacctcaagg agattctgac 4261 aacgggaagg aaaatcaaac cgtccataag aatcttgatg gattgttggg atcctaccca 4321 tacgacgtac ctgattatgc aacttatcct tatgatgtac ctgattatgc tagctatcct 4381 tatgatgtcc cagactacgc agccaccatg gtgagcaagg gcgaggagct gttcaccggg 4441 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc 4501 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc 4561 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc 4621 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa 4681 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc 4741 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc 4801 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc 4861 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac 4921 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac 4981 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac 5041 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact 5101 ctcggcatgg acgagctgta caagtaggga tccgcggccg cgaaggatct gcgatcgctc 5161 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg 5221 ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt 5281 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt 5341 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg 5401 ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt 5461 gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt 5521 aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag 5581 actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc 5641 gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagatgaccg 5701 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc 5761 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca 5821 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca 5881 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg 5941 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc 6001 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt 6061 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg 6121 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct 6181 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg 6241 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgagtcgaca 6301 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc 6361 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta 6421 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt 6481 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg 6541 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta 6601 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt 6661 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg 6721 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca 6781 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc 6841 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggtaccttta 6901 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga 6961 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct 7021 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 7081 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 7141 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt 7201 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt 7261 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 7321 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 7381 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt 7441 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct 7501 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg 7561 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 7621 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 7681 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 7741 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 7801 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 7861 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 7921 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 7981 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 8041 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 8101 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 8161 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 8221 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 8281 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 8341 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 8401 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 8461 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 8521 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 8581 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 8641 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 8701 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 8761 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 8821 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 8881 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 8941 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 9001 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 9061 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 9121 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 9181 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 9241 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 9301 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 9361 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 9421 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 9481 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 9541 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 9601 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 9661 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 9721 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 9781 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 9841 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 9901 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 9961 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga 10021 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat 10081 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat 10141 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca 10201 agctg //