LOCUS V024055 9545 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024055 VERSION V024055 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..3588 /label="Inhba(NM_008380)" /note="Inhba(NM_008380)" /gene="Inhba" CDS 3595..3660 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3704..3915 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3928..4196 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4225..4234 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4255..4917 /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 4987..5040 /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 5041..5640 /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 5641..6228 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6302..6535 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6607..6728 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6747..6882 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6920..6936) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6944..6960 /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(6968..6998) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7013..7034 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7322..7910) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8081..8941) /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(8942..9046) /gene="bla" /label="AmpR promoter" primer_bind 9520..9536 /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 gccaccatgc ccttgctttg gctgagagga 2341 tttctgttgg caagttgctg gattatagtg aggagttccc ccaccccagg atccgagggg 2401 cacggctcag ccccggactg cccgtcctgt gcgctggcca cccttccgaa ggatggacct 2461 aactctcagc cagagatggt agaggctgtc aagaagcaca tcttaaacat gctgcacttg 2521 aagaagagac ccgatgtcac ccagccggtg cccaaggcgg cgcttctcaa cgcgatcaga 2581 aagcttcatg tgggtaaagt gggggagaac gggtatgtgg agatagagga cgacattggc 2641 aggagggccg aaatgaatga actcatggag cagacctcgg agatcatcac ctttgccgag 2701 tcaggcacag ccaggaagac actgcacttt gagatttcca aggaaggcag tgacctgtca 2761 gtagtggagc gtgcagaagt gtggctcttc ctgaaagtcc ccaaggctaa cagaaccagg 2821 accaaagtca ccatccgtct atttcagcag cagaagcacc cacagggcag cttggacacg 2881 ggggatgagg ccgaggaaat gggcttaaag ggggagagga gtgaactgtt gctatcagag 2941 aaagtagttg atgctcggaa gagtacctgg cacatctttc cagtgtccag cagcatccag 3001 cgcctgctgg accagggaaa gagttccctg gacgtgcgga ttgcttgtga gcagtgccag 3061 gagagtggtg ccagtctagt gcttctgggc aagaagaaga agaaagaggt ggatggagat 3121 gggaagaaga aagatgggag tgacggaggg ctggaagagg aaaaggaaca gtcacataga 3181 cctttcctca tgctgcaggc taggcagtcc gaagaccacc ctcatcgcag gcgtaggcgg 3241 ggcttggagt gcgacggcaa ggtcaacatt tgctgtaaga aacagttctt tgtcagcttc 3301 aaggacattg gctggaatga ctggatcatt gctccctctg gctatcacgc caattattgt 3361 gagggggagt gcccaagcca catagcaggc acctctgggt cctcgctctc cttccactca 3421 acagtcatta accactaccg catgaggggt cacagcccct ttgccaacct taagtcatgc 3481 tgtgtgccca ccaagctgag acccatgtcc atgctgtatt acgatgatgg tcaaaacatc 3541 atcaaaaagg acattcaaaa catgattgtg gaggagtgtg gctgctccgg atccgactac 3601 aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag 3661 tagcggccgc gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg 3721 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg 3781 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt 3841 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt 3901 gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc 3961 cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt 4021 gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt 4081 tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc 4141 ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag 4201 ctgtgaccgg cgcctacgct agacgccacc atggagagcg acgagagcgg cctgcccgcc 4261 atggagatcg agtgccgcat caccggcacc ctgaacggcg tggagttcga gctggtgggc 4321 ggcggagagg gcacccccaa gcagggccgc atgaccaaca agatgaagag caccaaaggc 4381 gccctgacct tcagccccta cctgctgagc cacgtgatgg gctacggctt ctaccacttc 4441 ggcacctacc ccagcggcta cgagaacccc ttcctgcacg ccatcaacaa cggcggctac 4501 accaacaccc gcatcgagaa gtacgaggac ggcggcgtgc tgcacgtgag cttcagctac 4561 cgctacgagg ccggccgcgt gatcggcgac ttcaaggtgg tgggcaccgg cttccccgag 4621 gacagcgtga tcttcaccga caagatcatc cgcagcaacg ccaccgtgga gcacctgcac 4681 cccatgggcg ataacgtgct ggtgggcagc ttcgcccgca ccttcagcct gcgcgacggc 4741 ggctactaca gcttcgtggt ggacagccac atgcacttca agagcgccat ccaccccagc 4801 atcctgcaga acgggggccc catgttcgcc ttccgccgcg tggaggagct gcacagcaac 4861 accgagctgg gcatcgtgga gtaccagcac gccttcaaga cccccatcgc cttcgccaga 4921 tcccgcgctc agtcgtccaa ttctgccgtg gacggcaccg ccggacccgg ctccaccgga 4981 tctcgcgagg gcagaggaag tcttctaaca tgcggtgacg tggaggagaa tcccggccct 5041 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta 5101 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac 5161 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac 5221 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag 5281 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt 5341 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag 5401 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc 5461 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg 5521 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc 5581 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga 5641 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct 5701 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt 5761 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg 5821 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact 5881 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct 5941 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg 6001 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc 6061 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc 6121 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctc 6181 cgccttcgcc ctcagacgag tcggatctcc ctttggccgc ctccccgcct ggtaccttta 6241 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga 6301 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct 6361 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 6421 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 6481 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt 6541 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt 6601 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 6661 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 6721 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt 6781 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct 6841 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg 6901 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 6961 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 7021 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 7081 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 7141 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 7201 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 7261 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 7321 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 7381 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 7441 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 7501 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 7561 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 7621 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 7681 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 7741 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 7801 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 7861 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 7921 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 7981 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 8041 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 8101 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 8161 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 8221 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 8281 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 8341 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 8401 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 8461 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 8521 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 8581 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 8641 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 8701 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 8761 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 8821 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 8881 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 8941 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 9001 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 9061 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 9121 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 9181 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 9241 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 9301 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga 9361 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat 9421 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat 9481 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca 9541 agctg //