LOCUS V041428 8744 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V041428 VERSION V041428 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 2320..2337 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" CDS 2344..2838 /label="ISG15(NM_005101)" /note="ISG15(NM_005101)" /gene="ISG15" promoter 2903..3114 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3127..3395 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3424..3433 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3454..4116 /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 4186..4239 /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 4240..4839 /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 4840..5427 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5501..5734 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5806..5927 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5946..6081 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6119..6135) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6143..6159 /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(6167..6197) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6212..6233 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6521..7109) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7280..8140) /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(8141..8245) /gene="bla" /label="AmpR promoter" primer_bind 8719..8735 /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 accatcacca tcaccatctc 2341 gagatgggct gggacctgac ggtgaagatg ctggcgggca acgaattcca ggtgtccctg 2401 agcagctcca tgtcggtgtc agagctgaag gcgcagatca cccagaagat cggcgtgcac 2461 gccttccagc agcgtctggc tgtccacccg agcggtgtgg cgctgcagga cagggtcccc 2521 cttgccagcc agggcctggg ccccggcagc acggtcctgc tggtggtgga caaatgcgac 2581 gaacctctga gcatcctggt gaggaataac aagggccgca gcagcaccta cgaggtacgg 2641 ctgacgcaga ccgtggccca cctgaagcag caagtgagcg ggctggaggg tgtgcaggac 2701 gacctgttct ggctgacctt cgaggggaag cccctggagg accagctccc gctgggggag 2761 tacggcctca agcccctgag caccgtgttc atgaatctgc gcctgcgggg aggcggcaca 2821 gagcctggcg ggcggagcta aggatccctg cgatatccag cgcggccgcg aaggatctgc 2881 gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg 2941 gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa 3001 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt 3061 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acagctgaag 3121 cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg ccgccatcca 3181 cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc 3241 gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct cccttggagc 3301 ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca actctacgtc 3361 tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc gcctacgcta 3421 gacgccacca tggagagcga cgagagcggc ctgcccgcca tggagatcga gtgccgcatc 3481 accggcaccc tgaacggcgt ggagttcgag ctggtgggcg gcggagaggg cacccccaag 3541 cagggccgca tgaccaacaa gatgaagagc accaaaggcg ccctgacctt cagcccctac 3601 ctgctgagcc acgtgatggg ctacggcttc taccacttcg gcacctaccc cagcggctac 3661 gagaacccct tcctgcacgc catcaacaac ggcggctaca ccaacacccg catcgagaag 3721 tacgaggacg gcggcgtgct gcacgtgagc ttcagctacc gctacgaggc cggccgcgtg 3781 atcggcgact tcaaggtggt gggcaccggc ttccccgagg acagcgtgat cttcaccgac 3841 aagatcatcc gcagcaacgc caccgtggag cacctgcacc ccatgggcga taacgtgctg 3901 gtgggcagct tcgcccgcac cttcagcctg cgcgacggcg gctactacag cttcgtggtg 3961 gacagccaca tgcacttcaa gagcgccatc caccccagca tcctgcagaa cgggggcccc 4021 atgttcgcct tccgccgcgt ggaggagctg cacagcaaca ccgagctggg catcgtggag 4081 taccagcacg ccttcaagac ccccatcgcc ttcgccagat cccgcgctca gtcgtccaat 4141 tctgccgtgg acggcaccgc cggacccggc tccaccggat ctcgcgaggg cagaggaagt 4201 cttctaacat gcggtgacgt ggaggagaat cccggcccta tgaccgagta caagcccacg 4261 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc 4321 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc 4381 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg 4441 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg 4501 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa 4561 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc 4621 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga 4681 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac 4741 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga 4801 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgaa atcaacctct ggattacaaa 4861 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac 4921 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc 4981 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt 5041 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc 5101 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc 5161 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg 5221 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt 5281 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc 5341 cgcggcctgc tgccggctct gcggcctctt ccgcgtctcc gccttcgccc tcagacgagt 5401 cggatctccc tttggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca 5461 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc 5521 caacgaaaat aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga 5581 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 5641 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 5701 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt 5761 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca 5821 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 5881 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc 5941 tagctatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 6001 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt 6061 gaggaggctt ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt 6121 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 6181 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 6241 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 6301 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 6361 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 6421 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 6481 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 6541 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 6601 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 6661 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 6721 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 6781 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 6841 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 6901 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 6961 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 7021 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 7081 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 7141 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 7201 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 7261 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 7321 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 7381 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 7441 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 7501 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 7561 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 7621 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 7681 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 7741 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 7801 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 7861 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 7921 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 7981 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 8041 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 8101 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 8161 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 8221 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag 8281 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc 8341 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 8401 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 8461 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 8521 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 8581 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 8641 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 8701 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctg //