LOCUS V046018 9584 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V046018 VERSION V046018 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..3009 /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 3040..4509 /label="CLK3(NM_003992)" /note="CLK3(NM_003992)" /gene="CLK3" promoter 4554..4765 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4778..5046 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 5073..5672 /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 5679..6267 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6341..6574 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6646..6767 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6786..6921 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6959..6975) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6983..6999 /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(7007..7037) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7052..7073 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7361..7949) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8120..8980) /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(8981..9085) /gene="bla" /label="AmpR promoter" primer_bind 9559..9575 /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 catggtgagc aagggcgagg aggataacat ggccatcatc 2341 aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag 2401 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg 2461 accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc 2521 tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc 2581 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc 2641 caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac 2701 ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag 2761 cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag 2821 gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag 2881 ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac 2941 accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag 3001 ctgtacaagg gaggtggagg tagtggagga ggcggatcca tgcatcactg taagcgatac 3061 cgctcccctg aaccagaccc gtacctgagc taccgatgga agaggaggag gtcctacagt 3121 cgggaacatg aagggagact gcgatacccg tcccgaaggg agcctccccc acgaagatct 3181 cggtccagaa gccatgaccg cctgccctac cagaggaggt accgggagcg ccgtgacagc 3241 gatacatacc ggtgtgaaga gcggagccca tcctttggag aggactacta tggaccttca 3301 cgttctcgtc atcgtcggcg atcgcgggag agggggccat accggacccg caagcatgcc 3361 caccactgcc acaaacgccg caccaggtct tgtagcagcg cctcctcgag aagccaacag 3421 agcagtaagc gcagcagccg gagtgtggaa gatgacaagg agggtcacct ggtgtgccgg 3481 atcggcgatt ggctccaaga gcgatatgag attgtgggga acctgggtga aggcaccttt 3541 ggcaaggtgg tggagtgctt ggaccatgcc agagggaagt ctcaggttgc cctgaagatc 3601 atccgcaacg tgggcaagta ccgggaggct gcccggctag aaatcaacgt gctcaaaaaa 3661 atcaaggaga aggacaaaga aaacaagttc ctgtgtgtct tgatgtctga ctggttcaac 3721 ttccacggtc acatgtgcat cgcctttgag ctcctgggca agaacacctt tgagttcctg 3781 aaggagaata acttccagcc ttacccccta ccacatgtcc ggcacatggc ctaccagctc 3841 tgccacgccc ttagatttct gcatgagaat cagctgaccc atacagactt gaaaccagag 3901 aacatcctgt ttgtgaattc tgagtttgaa accctctaca atgagcacaa gagctgtgag 3961 gagaagtcag tgaagaacac cagcatccga gtggctgact ttggcagtgc cacatttgac 4021 catgagcacc acaccaccat tgtggccacc cgtcactatc gcccgcctga ggtgatcctt 4081 gagctgggct gggcacagcc ctgtgacgtc tggagcattg gctgcattct ctttgagtac 4141 taccggggct tcacactctt ccagacccac gaaaaccgag agcacctggt gatgatggag 4201 aagatcctag ggcccatccc atcacacatg atccaccgta ccaggaagca gaaatatttc 4261 tacaaagggg gcctagtttg ggatgagaac agctctgacg gccggtatgt gaaggagaac 4321 tgcaaacctc tgaagagtta catgctccaa gactccctgg agcacgtgca gctgtttgac 4381 ctgatgagga ggatgttaga atttgaccct gcccagcgca tcacactggc cgaggccctg 4441 ctgcacccct tctttgctgg cctgacccct gaggagcggt ccttccacac cagccgcaac 4501 ccaagcagat gagcggccgc gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga 4561 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc 4621 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt 4681 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg 4741 caacgggttt gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg 4801 cgcccgccgc cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc 4861 cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag 4921 accgggcctt tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct 4981 ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta 5041 cagatccaag ctgtgaccgg cgcctacgct agatgaccga gtacaagccc acggtgcgcc 5101 tcgccacccg cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg ttcgccgact 5161 accccgccac gcgccacacc gtcgacccgg accgccacat cgagcgggtc accgagctgc 5221 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg 5281 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg 5341 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg 5401 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg 5461 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg 5521 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct 5581 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca 5641 cctggtgcat gacccgcaag cccggtgcct gagtcgacaa tcaacctctg gattacaaaa 5701 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg 5761 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct 5821 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg 5881 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct 5941 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg 6001 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg 6061 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc 6121 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc 6181 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc 6241 ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca 6301 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc 6361 caacgaaaat aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga 6421 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 6481 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 6541 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt 6601 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca 6661 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 6721 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc 6781 tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 6841 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt 6901 gaggaggctt ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt 6961 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 7021 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 7081 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 7141 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 7201 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 7261 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 7321 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 7381 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 7441 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 7501 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 7561 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 7621 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 7681 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 7741 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 7801 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 7861 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 7921 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 7981 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 8041 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 8101 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 8161 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 8221 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 8281 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 8341 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 8401 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 8461 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 8521 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 8581 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 8641 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 8701 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 8761 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 8821 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 8881 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 8941 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 9001 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 9061 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag 9121 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc 9181 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 9241 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 9301 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 9361 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 9421 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 9481 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 9541 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctg //