LOCUS V034066 8484 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V034066 VERSION V034066 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..1921 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2011..2214 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2293..2565 /label="Ccl5(NM_013653)" /note="Ccl5(NM_013653)" /gene="Ccl5" CDS 2569..2598 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /transl_table=1 /translation="EQKLISEEDL" /label="Myc" promoter 2643..2854 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 2867..3135 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3164..3173 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3194..3856 /codon_start=1 /product="green fluorescent protein 2 from Pontellina plumata, also known as ppluGFP2 (Shagin et al., 2004)" /transl_table=1 /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA" /label="CopGFP" CDS 3926..3979 /codon_start=1 /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /product="2A peptide from Thosea asigna virus capsid protein" /protein_id="" /transl_table=1 /translation="EGRGSLLTCGDVEENPGP" /label="T2A" CDS 3980..4579 /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 4580..5167 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5241..5474 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5546..5667 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5686..5821 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5859..5875) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5883..5899 /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(5907..5937) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5952..5973 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6261..6849) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7020..7880) /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(7881..7985) /gene="bla" /label="AmpR promoter" primer_bind 8459..8475 /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 attacaaaaa ttcaaaattt 1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac 1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg 2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg 2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca 2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta 2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc 2281 tagagctagc gaatgaagat ctctgcagct gccctcacca tcatcctcac tgcagccgcc 2341 ctctgcaccc ccgcacctgc ctcaccatat ggctcggaca ccactccctg ctgctttgcc 2401 tacctctccc tcgcgctgcc tcgtgcccac gtcaaggagt atttctacac cagcagcaag 2461 tgctccaatc ttgcagtcgt gtttgtcact cgaaggaacc gccaagtgtg tgccaaccca 2521 gagaagaagt gggttcaaga atacatcaac tatttggaga tgagctccga acaaaaactc 2581 atctcagaag aggatctgtg agcggccgcg aaggatctgc gatcgctccg gtgcccgtca 2641 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg 2701 aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct 2761 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt 2821 tctttttcgc aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc 2881 tccttcacgc gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct 2941 gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct 3001 caggtcgaga ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct 3061 ctccacgctt tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct 3121 gcgccgttac agatccaagc tgtgaccggc gcctacgcta gacgccacca tggagagcga 3181 cgagagcggc ctgcccgcca tggagatcga gtgccgcatc accggcaccc tgaacggcgt 3241 ggagttcgag ctggtgggcg gcggagaggg cacccccaag cagggccgca tgaccaacaa 3301 gatgaagagc accaaaggcg ccctgacctt cagcccctac ctgctgagcc acgtgatggg 3361 ctacggcttc taccacttcg gcacctaccc cagcggctac gagaacccct tcctgcacgc 3421 catcaacaac ggcggctaca ccaacacccg catcgagaag tacgaggacg gcggcgtgct 3481 gcacgtgagc ttcagctacc gctacgaggc cggccgcgtg atcggcgact tcaaggtggt 3541 gggcaccggc ttccccgagg acagcgtgat cttcaccgac aagatcatcc gcagcaacgc 3601 caccgtggag cacctgcacc ccatgggcga taacgtgctg gtgggcagct tcgcccgcac 3661 cttcagcctg cgcgacggcg gctactacag cttcgtggtg gacagccaca tgcacttcaa 3721 gagcgccatc caccccagca tcctgcagaa cgggggcccc atgttcgcct tccgccgcgt 3781 ggaggagctg cacagcaaca ccgagctggg catcgtggag taccagcacg ccttcaagac 3841 ccccatcgcc ttcgccagat cccgcgctca gtcgtccaat tctgccgtgg acggcaccgc 3901 cggacccggc tccaccggat ctcgcgaggg cagaggaagt cttctaacat gcggtgacgt 3961 ggaggagaat cccggcccta tgaccgagta caagcccacg gtgcgcctcg ccacccgcga 4021 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg 4081 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct 4141 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc 4201 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg 4261 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc 4321 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca 4381 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc 4441 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct 4501 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac 4561 ccgcaagccc ggtgcctgaa atcaacctct ggattacaaa atttgtgaaa gattgactgg 4621 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta 4681 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct 4741 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt 4801 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac 4861 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg 4921 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac 4981 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg 5041 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct 5101 gcggcctctt ccgcgtctcc gccttcgccc tcagacgagt cggatctccc tttggccgcc 5161 tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt 5221 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaaaat aagatctgct 5281 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta 5341 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg 5401 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg 5461 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa 5521 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata 5581 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg 5641 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact 5701 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 5761 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 5821 ctagactttt gcagagacgg cccaaattcg taatcatggt catagctgtt tcctgtgtga 5881 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc 5941 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc 6001 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 6061 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 6121 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 6181 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 6241 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 6301 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 6361 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 6421 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 6481 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 6541 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 6601 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 6661 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc 6721 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 6781 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 6841 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 6901 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 6961 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 7021 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 7081 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 7141 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 7201 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 7261 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 7321 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 7381 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 7441 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 7501 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 7561 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 7621 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 7681 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 7741 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 7801 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 7861 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 7921 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 7981 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca 8041 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac 8101 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat 8161 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg 8221 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata 8281 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc 8341 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg 8401 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt 8461 aaaacgacgg ccagtgccaa gctg //