LOCUS V040923 9434 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040923 VERSION V040923 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..3435 /label="NSDHL(NM_015922)" /note="NSDHL(NM_015922)" /gene="NSDHL" CDS 3442..3468 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3472..3498 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3502..3528 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" promoter 3593..3804 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3817..4085 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4114..4123 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4144..4806 /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 4876..4929 /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 4930..5529 /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 5530..6117 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6191..6424 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6496..6617 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6636..6771 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6809..6825) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6833..6849 /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(6857..6887) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6902..6923 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7211..7799) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7970..8830) /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(8831..8935) /gene="bla" /label="AmpR promoter" primer_bind 9409..9425 /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 gccaccatgg aaccagcagt tagcgagcca 2341 atgagagacc aagtcgcacg gactcatttg acagaggaca ctcccaaagt gaatgctgac 2401 atagaaaagg ttaaccagaa tcaggccaag agatgcacag tgatcggtgg ctctggattc 2461 ctggggcagc acatggtgga gcagttgctg gcaagaggat atgctgtcaa tgtatttgat 2521 atccagcaag ggtttgataa tccccaggtg cggttctttc tgggtgacct ctgcagccga 2581 caggatctgt acccagctct gaaaggtgta aacacagttt tccactgtgc gtcaccccca 2641 ccatccagta acaacaagga gctcttttat agagtgaatt acattggcac caagaatgtc 2701 attgaaactt gcaaagaggc tggggttcag aaactcattt taaccagcag tgccagtgtc 2761 atctttgagg gcgtcgatat caagaatgga actgaagacc ttccctatgc catgaaaccc 2821 attgactact acacagagac taagatctta caggagaggg cagttctggg cgccaacgat 2881 cctgagaaga atttcttaac cacagccatc cgccctcatg gcattttcgg cccaagggac 2941 ccgcagttgg tacccatcct catcgaggca gccaggaacg gcaagatgaa gttcgtgatt 3001 ggaaatggga agaacttggt ggacttcacc tttgtggaga acgtggtcca tggacacatc 3061 ctggcggcag agcagctctc ccgagactcg acactgggtg ggaaggcatt tcacatcacc 3121 aatgatgagc ccatcccttt ctggacattc ctgtctcgca tcctgacagg cctcaattat 3181 gaggccccca agtaccacat cccctactgg gtggcctact acctggccct cctgctatcc 3241 ctgctggtga tggtgatcag tcctgtcatc cagctgcagc ccaccttcac acccatgcgg 3301 gtcgcactgg ctggcacatt ccactactac agctgcgaga gagccaaaaa ggccatgggc 3361 taccagccac tagtgaccat ggatgatgct atggagagga ccgtgcagag ctttcgccac 3421 ctgcggaggg tcaagctcga gtacccatac gacgtacctg attatgcaac ttatccttat 3481 gatgtacctg attatgctag ctatccttat gatgtcccag actacgcata gggatccctg 3541 cgatatccag cgcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag 3601 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc 3661 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt 3721 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc 3781 aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc 3841 gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc 3901 gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga 3961 ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt 4021 tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac 4081 agatccaagc tgtgaccggc gcctacgcta gacgccacca tggagagcga cgagagcggc 4141 ctgcccgcca tggagatcga gtgccgcatc accggcaccc tgaacggcgt ggagttcgag 4201 ctggtgggcg gcggagaggg cacccccaag cagggccgca tgaccaacaa gatgaagagc 4261 accaaaggcg ccctgacctt cagcccctac ctgctgagcc acgtgatggg ctacggcttc 4321 taccacttcg gcacctaccc cagcggctac gagaacccct tcctgcacgc catcaacaac 4381 ggcggctaca ccaacacccg catcgagaag tacgaggacg gcggcgtgct gcacgtgagc 4441 ttcagctacc gctacgaggc cggccgcgtg atcggcgact tcaaggtggt gggcaccggc 4501 ttccccgagg acagcgtgat cttcaccgac aagatcatcc gcagcaacgc caccgtggag 4561 cacctgcacc ccatgggcga taacgtgctg gtgggcagct tcgcccgcac cttcagcctg 4621 cgcgacggcg gctactacag cttcgtggtg gacagccaca tgcacttcaa gagcgccatc 4681 caccccagca tcctgcagaa cgggggcccc atgttcgcct tccgccgcgt ggaggagctg 4741 cacagcaaca ccgagctggg catcgtggag taccagcacg ccttcaagac ccccatcgcc 4801 ttcgccagat cccgcgctca gtcgtccaat tctgccgtgg acggcaccgc cggacccggc 4861 tccaccggat ctcgcgaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat 4921 cccggcccta tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc 4981 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc 5041 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc 5101 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc 5161 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag 5221 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg 5281 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag 5341 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc 5401 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc 5461 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc 5521 ggtgcctgaa atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac 5581 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt 5641 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat 5701 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca 5761 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc 5821 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg 5881 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca 5941 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct 6001 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt 6061 ccgcgtctcc gccttcgccc tcagacgagt cggatctccc tttggccgcc tccccgcctg 6121 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa 6181 aaggggggac tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt 6241 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac 6301 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg 6361 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct 6421 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata 6481 tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc 6541 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa 6601 ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt 6661 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 6721 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt 6781 gcagagacgg cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc 6841 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct 6901 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa 6961 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta 7021 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc 7081 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg 7141 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt 7201 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa 7261 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct 7321 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 7381 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg 7441 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 7501 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 7561 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 7621 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga 7681 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 7741 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 7801 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 7861 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 7921 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 7981 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 8041 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 8101 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 8161 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 8221 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 8281 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 8341 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 8401 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 8461 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 8521 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 8581 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 8641 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 8701 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 8761 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 8821 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 8881 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 8941 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata 9001 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc 9061 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca 9121 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg 9181 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat 9241 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg 9301 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg 9361 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg 9421 ccagtgccaa gctg //