LOCUS V026114 9844 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V026114 VERSION V026114 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 2311..3870 /label="MARCO(NM_006770)" /note="MARCO(NM_006770)" /gene="MARCO" CDS 3877..3942 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3986..4197 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4210..4478 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4507..4516 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4537..5199 /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 5269..5322 /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 5323..5922 /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 5923..6511 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6585..6818 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6890..7011 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7051..7186 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7219..7235) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7243..7259 /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(7267..7297) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7312..7333 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7621..8209) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8380..9240) /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(9241..9345) /gene="bla" /label="AmpR promoter" primer_bind 9819..9835 /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 ctgatacgaa ctcggaattc atgagaaata agaaaattct caaggaggac 2341 gagctcttga gtgagaccca acaagctgct tttcaccaaa ttgcaatgga gcctttcgaa 2401 atcaatgttc caaagcccaa gaggagaaat ggggtgaact tctccctagc tgtggtggtc 2461 atctacctga tcctgctcac cgctggcgct gggctgctgg tggtccaagt tctgaatctg 2521 caggcgcggc tccgggtcct ggagatgtat ttcctcaatg acactctggc ggctgaggac 2581 agcccgtcct tctccttgct gcagtcagca caccctggag aacacctggc tcagggtgca 2641 tcgaggctgc aagtcctgca ggcccaactc acctgggtcc gcgtcagcca tgagcacttg 2701 ctgcagcggg tagacaactt cactcagaac ccagggatgt tcagaatcaa aggtgaacaa 2761 ggcgccccag gtcttcaagg tcacaagggg gccatgggca tgcctggtgc ccctggcccg 2821 ccgggaccac ctgctgagaa gggagccaag ggggctatgg gacgagatgg agcaacaggc 2881 ccctcgggac cccaaggccc accgggagtc aagggagagg cgggcctcca aggaccccag 2941 ggtgctccag ggaagcaagg agccactggc accccaggac cccaaggaga gaagggcagc 3001 aaaggcgatg ggggtctcat tggcccaaaa ggggaaactg gaactaaggg agagaaagga 3061 gacctgggtc tcccaggaag caaaggggac aggggcatga aaggagatgc aggggtcatg 3121 gggcctcctg gagcccaggg gagtaaaggt gacttcggga ggccaggccc accaggtttg 3181 gctggttttc ctggagctaa aggagatcaa ggacaacctg gactgcaggg tgttccgggc 3241 cctcctggtg cagtgggaca cccaggtgcc aagggtgagc ctggcagtgc tggctcccct 3301 gggcgagcag gacttccagg gagccccggg agtccaggag ccacaggcct gaaaggaagc 3361 aaaggggaca caggacttca aggacagcaa ggaagaaaag gagaatcagg agttccaggc 3421 cctgcaggtg tgaagggaga acaggggagc ccagggctgg caggtcccaa gggagcccct 3481 ggacaagctg gccagaaggg agaccaggga gtgaaaggat cttctgggga gcaaggagta 3541 aagggagaaa aaggtgaaag aggtgaaaac tcagtgtccg tcaggattgt cggcagtagt 3601 aaccgaggcc gggctgaagt ttactacagt ggtacctggg ggacaatttg cgatgacgag 3661 tggcaaaatt ctgatgccat tgtcttctgc cgcatgctgg gttactccaa aggaagggcc 3721 ctgtacaaag tgggagctgg cactgggcag atctggctgg ataatgttca gtgtcggggc 3781 acggagagta ccctgtggag ctgcaccaag aatagctggg gccatcatga ctgcagccac 3841 gaggaggacg caggcgtgga gtgcagcgtc ggatccgact acaaagacca tgacggtgat 3901 tataaagatc atgacatcga ctacaaggat gacgatgaca agtagcggcc gcgaaggatc 3961 tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag 4021 ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg 4081 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata 4141 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg 4201 aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat 4261 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc 4321 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg 4381 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac 4441 gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg 4501 ctagacgcca ccatggagag cgacgagagc ggcctgcccg ccatggagat cgagtgccgc 4561 atcaccggca ccctgaacgg cgtggagttc gagctggtgg gcggcggaga gggcaccccc 4621 aagcagggcc gcatgaccaa caagatgaag agcaccaaag gcgccctgac cttcagcccc 4681 tacctgctga gccacgtgat gggctacggc ttctaccact tcggcaccta ccccagcggc 4741 tacgagaacc ccttcctgca cgccatcaac aacggcggct acaccaacac ccgcatcgag 4801 aagtacgagg acggcggcgt gctgcacgtg agcttcagct accgctacga ggccggccgc 4861 gtgatcggcg acttcaaggt ggtgggcacc ggcttccccg aggacagcgt gatcttcacc 4921 gacaagatca tccgcagcaa cgccaccgtg gagcacctgc accccatggg cgataacgtg 4981 ctggtgggca gcttcgcccg caccttcagc ctgcgcgacg gcggctacta cagcttcgtg 5041 gtggacagcc acatgcactt caagagcgcc atccacccca gcatcctgca gaacgggggc 5101 cccatgttcg ccttccgccg cgtggaggag ctgcacagca acaccgagct gggcatcgtg 5161 gagtaccagc acgccttcaa gacccccatc gccttcgcca gatcccgcgc tcagtcgtcc 5221 aattctgccg tggacggcac cgccggaccc ggctccaccg gatctcgcga gggcagagga 5281 agtcttctaa catgcggtga cgtggaggag aatcccggcc ctatgaccga gtacaagccc 5341 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg 5401 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc 5461 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc 5521 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg 5581 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag 5641 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc 5701 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc 5761 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc 5821 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa 5881 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaaatcaacc tctggattac 5941 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 6001 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 6061 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 6121 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 6181 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 6241 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 6301 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg 6361 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 6421 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc tccgccttcg ccctcagacg 6481 agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa tgacttacaa 6541 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca 6601 ctcccaacga agataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat 6661 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt 6721 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc 6781 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat 6841 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat 6901 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 6961 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 7021 gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 7081 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 7141 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctaga cttttgcaga 7201 gaccaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat 7261 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 7321 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 7381 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 7441 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 7501 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 7561 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 7621 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 7681 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 7741 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 7801 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 7861 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 7921 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 7981 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 8041 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 8101 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 8161 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 8221 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 8281 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 8341 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 8401 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 8461 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 8521 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 8581 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 8641 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 8701 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 8761 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 8821 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 8881 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 8941 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 9001 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 9061 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 9121 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 9181 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 9241 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 9301 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 9361 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 9421 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 9481 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 9541 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 9601 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 9661 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 9721 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 9781 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 9841 gctg //