LOCUS V023553 10025 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V023553 VERSION V023553 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 2317..4068 /label="ME2(NM_002396)" /note="ME2(NM_002396)" /gene="ME2" CDS 4075..4140 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4184..4395 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4408..4676 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4705..4714 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4735..5397 /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 5467..5520 /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 5521..6120 /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 6121..6708 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6782..7015 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7087..7208 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7227..7362 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7400..7416) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7424..7440 /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(7448..7478) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7493..7514 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7802..8390) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8561..9421) /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(9422..9526) /gene="bla" /label="AmpR promoter" primer_bind 10000..10016 /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 gccaccatgt tgtcccggtt aagagtagtt 2341 tccaccactt gtactttggc atgtcgacat ttgcacataa aagaaaaagg caagccactt 2401 atgctgaacc caagaacaaa caagggaatg gcatttactt tacaagaacg acaaatgctt 2461 ggtcttcaag gacttctacc tcccaaaata gagacacaag atattcaagc cttacgattt 2521 catagaaact tgaagaaaat gactagccct ttggaaaaat atatctacat aatgggaata 2581 caagaaagaa atgagaaatt gttttataga atactgcaag atgacattga gagtttaatg 2641 ccaattgtat atacaccgac ggttggtctt gcctgctccc agtatggaca catctttaga 2701 agacctaagg gattatttat ttcgatctca gacagaggtc atgttagatc aattgtggat 2761 aactggccag aaaatcatgt taaggctgtt gtagtgactg atggagagag aattctgggt 2821 cttggagatc tgggtgtcta tggaatggga attccagtag gaaaactttg tttgtataca 2881 gcttgtgcag gaatacggcc tgatagatgc ctgccagtgt gtattgatgt gggaactgat 2941 aatatcgcac tcttaaaaga cccattttac atgggcttgt accagaaacg agatcgcaca 3001 caacagtatg atgacctgat tgatgagttt atgaaagcta ttactgacag atatggccgg 3061 aacacactca ttcagttcga agactttgga aatcataatg cattcaggtt cttgagaaag 3121 taccgagaaa aatattgtac tttcaatgat gatattcaag ggacagctgc agtagctcta 3181 gcaggtcttc ttgcagcaca aaaagttatt agtaaaccaa tctccgaaca caaaatctta 3241 ttccttggag caggagaggc tgctcttgga attgcaaatc ttatagttat gtctatggta 3301 gaaaatggcc tgtcagaaca agaggcacaa aagaaaatct ggatgtttga caagtatggt 3361 ttattagtta agggacggaa agcaaaaata gatagttatc aggaaccatt tactcactca 3421 gccccagaga gcatacctga tacttttgaa gatgcagtga atatactgaa gccttcaact 3481 ataattggag ttgcaggtgc tggccgtctt ttcactcctg atgtaatcag agccatggcc 3541 tctatcaatg aaaggcctgt aatatttgca ttaagtaatc ctacagcaca ggcagagtgc 3601 acggctgaag aagcatatac acttacagag ggcaggtgtt tgtttgccag tggcagtcca 3661 tttgggccag tgaaacttac agatgggcga gtctttacac caggtcaagg aaacaatgtt 3721 tatatttttc caggtgtggc tttagctgtt attctctgta acacccggca tattagtgac 3781 agtgttttcc tagaagctgc aaaggccctg acaagccaat tgacagatga agagctagcc 3841 caagggagac tttacccacc gcttgctaat attcaggaag tttctattaa cattgctatt 3901 aaagttacag aatacctata tgctaataaa atggctttcc gatacccaga acctgaagac 3961 aaggccaaat atgttaaaga aagaacatgg cggagtgaat atgattccct gctgccagat 4021 gtgtatgaat ggccagaatc tgcatcaagc cctcctgtga taacagaagg atccgactac 4081 aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag 4141 tagcggccgc gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg 4201 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg 4261 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt 4321 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt 4381 gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc 4441 cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt 4501 gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt 4561 tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc 4621 ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag 4681 ctgtgaccgg cgcctacgct agacgccacc atggagagcg acgagagcgg cctgcccgcc 4741 atggagatcg agtgccgcat caccggcacc ctgaacggcg tggagttcga gctggtgggc 4801 ggcggagagg gcacccccaa gcagggccgc atgaccaaca agatgaagag caccaaaggc 4861 gccctgacct tcagccccta cctgctgagc cacgtgatgg gctacggctt ctaccacttc 4921 ggcacctacc ccagcggcta cgagaacccc ttcctgcacg ccatcaacaa cggcggctac 4981 accaacaccc gcatcgagaa gtacgaggac ggcggcgtgc tgcacgtgag cttcagctac 5041 cgctacgagg ccggccgcgt gatcggcgac ttcaaggtgg tgggcaccgg cttccccgag 5101 gacagcgtga tcttcaccga caagatcatc cgcagcaacg ccaccgtgga gcacctgcac 5161 cccatgggcg ataacgtgct ggtgggcagc ttcgcccgca ccttcagcct gcgcgacggc 5221 ggctactaca gcttcgtggt ggacagccac atgcacttca agagcgccat ccaccccagc 5281 atcctgcaga acgggggccc catgttcgcc ttccgccgcg tggaggagct gcacagcaac 5341 accgagctgg gcatcgtgga gtaccagcac gccttcaaga cccccatcgc cttcgccaga 5401 tcccgcgctc agtcgtccaa ttctgccgtg gacggcaccg ccggacccgg ctccaccgga 5461 tctcgcgagg gcagaggaag tcttctaaca tgcggtgacg tggaggagaa tcccggccct 5521 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta 5581 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac 5641 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac 5701 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag 5761 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt 5821 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag 5881 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc 5941 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg 6001 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc 6061 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga 6121 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct 6181 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt 6241 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg 6301 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact 6361 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct 6421 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg 6481 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc 6541 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc 6601 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctc 6661 cgccttcgcc ctcagacgag tcggatctcc ctttggccgc ctccccgcct ggtaccttta 6721 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga 6781 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct 6841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 6901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 6961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt 7021 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt 7081 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 7141 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 7201 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt 7261 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct 7321 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg 7381 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 7441 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 7501 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 7561 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 7621 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 7681 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 7741 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 7801 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 7861 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 7921 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 7981 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 8041 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 8101 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 8161 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 8221 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 8281 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 8341 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 8401 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 8461 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 8521 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 8581 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 8641 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 8701 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 8761 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 8821 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 8881 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 8941 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 9001 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 9061 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 9121 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 9181 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 9241 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 9301 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 9361 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 9421 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 9481 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 9541 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 9601 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 9661 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 9721 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 9781 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga 9841 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat 9901 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat 9961 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca 10021 agctg //