LOCUS V020133 10265 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020133 VERSION V020133 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 1798..1915 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2005..2208 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2291..4375 /label="APP(NM_201414)" /note="APP(NM_201414)" /gene="APP" promoter 4424..4635 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4648..4916 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4945..4954 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4975..5637 /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 5707..5760 /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 5761..6360 /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 6361..6948 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 7022..7255 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7327..7448 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7467..7602 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7640..7656) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7664..7680 /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(7688..7718) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7733..7754 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(8042..8630) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8801..9661) /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(9662..9766) /gene="bla" /label="AmpR promoter" primer_bind 10240..10256 /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 ggttaacttt 1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc 1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga 1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac 1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga 2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg 2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg 2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac 2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc 2281 tagcgccacc atgctgcccg gtttggcact gctcctgctg gccgcctgga cggctcgggc 2341 gctggaggta cccactgatg gtaatgctgg cctgctggct gaaccccaga ttgccatgtt 2401 ctgtggcaga ctgaacatgc acatgaatgt ccagaatggg aagtgggatt cagatccatc 2461 agggaccaaa acctgcattg ataccaagga aggcatcctg cagtattgcc aagaagtcta 2521 ccctgaactg cagatcacca atgtggtaga agccaaccaa ccagtgacca tccagaactg 2581 gtgcaagcgg ggccgcaagc agtgcaagac ccatccccac tttgtgattc cctaccgctg 2641 cttagttggt gagtttgtaa gtgatgccct tctcgttcct gacaagtgca aattcttaca 2701 ccaggagagg atggatgttt gcgaaactca tcttcactgg cacaccgtcg ccaaagagac 2761 atgcagtgag aagagtacca acttgcatga ctacggcatg ttgctgccct gcggaattga 2821 caagttccga ggggtagagt ttgtgtgttg cccactggct gaagaaagtg acaatgtgga 2881 ttctgctgat gcggaggagg atgactcgga tgtctggtgg ggcggagcag acacagacta 2941 tgcagatggg agtgaagaca aagtagtaga agtagcagag gaggaagaag tggctgaggt 3001 ggaagaagaa gaagccgatg atgacgagga cgatgaggat ggtgatgagg tagaggaaga 3061 ggctgaggaa ccctacgaag aagccacaga gagaaccacc agcattgcca ccaccaccac 3121 caccaccaca gagtctgtgg aagaggtggt tcgagttcct acaacagcag ccagtacccc 3181 tgatgccgtt gacaagtatc tcgagacacc tggggatgag aatgaacatg cccatttcca 3241 gaaagccaaa gagaggcttg aggccaagca ccgagagaga atgtcccagg tcatgagaga 3301 atgggaagag gcagaacgtc aagcaaagaa cttgcctaaa gctgataaga aggcagttat 3361 ccagcatttc caggagaaag tggaatcttt ggaacaggaa gcagccaacg agagacagca 3421 gctggtggag acacacatgg ccagagtgga agccatgctc aatgaccgcc gccgcctggc 3481 cctggagaac tacatcaccg ctctgcaggc tgttcctcct cggcctcgtc acgtgttcaa 3541 tatgctaaag aagtatgtcc gcgcagaaca gaaggacaga cagcacaccc taaagcattt 3601 cgagcatgtg cgcatggtgg atcccaagaa agccgctcag atccggtccc aggttatgac 3661 acacctccgt gtgatttatg agcgcatgaa tcagtctctc tccctgctct acaacgtgcc 3721 tgcagtggcc gaggagattc aggatgaagt tgatgagctg cttcagaaag agcaaaacta 3781 ttcagatgac gtcttggcca acatgattag tgaaccaagg atcagttacg gaaacgatgc 3841 tctcatgcca tctttgaccg aaacgaaaac caccgtggag ctccttcccg tgaatggaga 3901 gttcagcctg gacgatctcc agccgtggca ttcttttggg gctgactctg tgccagccaa 3961 cacagaaaac gaagttgagc ctgttgatgc ccgccctgct gccgaccgag gactgaccac 4021 tcgaccaggt tctgggttga caaatatcaa gacggaggag atctctgaag tgaatctgga 4081 tgcagaattc cgacatgact caggatatga agttcatcat caaaaattgg tgttctttgc 4141 agaagatgtg ggttcaaaca aaggtgcaat cattggactc atggtgggcg gtgttgtcat 4201 agcgacagtg atcgtcatca ccttggtgat gctgaagaag aaacagtaca catccattca 4261 tcatggtgtg gtggaggttg acgccgctgt caccccagag gagcgccacc tgtccaagat 4321 gcagcagaac ggctacgaaa atccaaccta caagttcttt gagcagatgc agaactagga 4381 tccgcggccg caaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg 4441 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg 4501 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt 4561 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt 4621 gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc 4681 cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt 4741 gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt 4801 tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc 4861 ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag 4921 ctgtgaccgg cgcctacgct agacgccacc atggagagcg acgagagcgg cctgcccgcc 4981 atggagatcg agtgccgcat caccggcacc ctgaacggcg tggagttcga gctggtgggc 5041 ggcggagagg gcacccccaa gcagggccgc atgaccaaca agatgaagag caccaaaggc 5101 gccctgacct tcagccccta cctgctgagc cacgtgatgg gctacggctt ctaccacttc 5161 ggcacctacc ccagcggcta cgagaacccc ttcctgcacg ccatcaacaa cggcggctac 5221 accaacaccc gcatcgagaa gtacgaggac ggcggcgtgc tgcacgtgag cttcagctac 5281 cgctacgagg ccggccgcgt gatcggcgac ttcaaggtgg tgggcaccgg cttccccgag 5341 gacagcgtga tcttcaccga caagatcatc cgcagcaacg ccaccgtgga gcacctgcac 5401 cccatgggcg ataacgtgct ggtgggcagc ttcgcccgca ccttcagcct gcgcgacggc 5461 ggctactaca gcttcgtggt ggacagccac atgcacttca agagcgccat ccaccccagc 5521 atcctgcaga acgggggccc catgttcgcc ttccgccgcg tggaggagct gcacagcaac 5581 accgagctgg gcatcgtgga gtaccagcac gccttcaaga cccccatcgc cttcgccaga 5641 tcccgcgctc agtcgtccaa ttctgccgtg gacggcaccg ccggacccgg ctccaccgga 5701 tctcgcgagg gcagaggaag tcttctaaca tgcggtgacg tggaggagaa tcccggccct 5761 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta 5821 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac 5881 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac 5941 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag 6001 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt 6061 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag 6121 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc 6181 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg 6241 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc 6301 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga 6361 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct 6421 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt 6481 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg 6541 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact 6601 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct 6661 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg 6721 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc 6781 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc 6841 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctc 6901 cgccttcgcc ctcagacgag tcggatctcc ctttggccgc ctccccgcct ggtaccttta 6961 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga 7021 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct 7081 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 7141 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 7201 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt 7261 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt 7321 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 7381 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 7441 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt 7501 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct 7561 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg 7621 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 7681 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 7741 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 7801 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 7861 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 7921 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 7981 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 8041 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 8101 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 8161 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 8221 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 8281 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 8341 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 8401 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 8461 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 8521 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 8581 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 8641 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 8701 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 8761 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 8821 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 8881 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 8941 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 9001 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 9061 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 9121 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 9181 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 9241 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 9301 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 9361 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 9421 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 9481 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 9541 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 9601 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 9661 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 9721 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 9781 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 9841 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 9901 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 9961 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 10021 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga 10081 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat 10141 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat 10201 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca 10261 agctg //