LOCUS V038318 8824 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V038318 VERSION V038318 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..2850 /label="MED30(NM_080651)" /note="MED30(NM_080651)" /gene="MED30" CDS 2857..2922 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 2966..3177 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3190..3458 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3487..3496 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3517..4179 /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 4249..4302 /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 4303..4902 /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 4903..5491 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5565..5798 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5870..5991 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6031..6166 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6199..6215) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6223..6239 /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(6247..6277) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6292..6313 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6601..7189) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7360..8220) /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(8221..8325) /gene="bla" /label="AmpR promoter" primer_bind 8799..8815 /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 ccacccctcc gttggccgcg 2341 tcggggatgg cgcccgggcc cttcgccggg ccccaggctc agcaggccgc ccgggaagtc 2401 aacacggcgt cgctgtgccg catcgggcag gagacagtgc aggacatcgt gtaccgcacc 2461 atggagatct tccagctcct gaggaacatg cagctgccaa atggtgtcac ttaccacact 2521 ggaacatatc aagaccggtt aacaaagcta caggataatc ttcgccaact ttcagttctc 2581 ttcaggaagc tgagattggt atatgacaaa tgcaatgaaa actgtggtgg gatggatccc 2641 attccagtcg agcaacttat tccatatgtg gaagaagatg gctcaaagaa tgatgatcgg 2701 gctggcccac ctcgttttgc tagtgaagag aggcgagaaa ttgctgaagt aaataaaaaa 2761 ctcaaacaga agaatcaaca gctgaaacaa attatggatc aattacgaaa tctcatctgg 2821 gatataaatg ccatgttggc aatgaggaac ggatccgact acaaagacca tgacggtgat 2881 tataaagatc atgacatcga ctacaaggat gacgatgaca agtagcggcc gcgaaggatc 2941 tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag 3001 ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg 3061 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata 3121 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg 3181 aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat 3241 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc 3301 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg 3361 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac 3421 gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg 3481 ctagacgcca ccatggagag cgacgagagc ggcctgcccg ccatggagat cgagtgccgc 3541 atcaccggca ccctgaacgg cgtggagttc gagctggtgg gcggcggaga gggcaccccc 3601 aagcagggcc gcatgaccaa caagatgaag agcaccaaag gcgccctgac cttcagcccc 3661 tacctgctga gccacgtgat gggctacggc ttctaccact tcggcaccta ccccagcggc 3721 tacgagaacc ccttcctgca cgccatcaac aacggcggct acaccaacac ccgcatcgag 3781 aagtacgagg acggcggcgt gctgcacgtg agcttcagct accgctacga ggccggccgc 3841 gtgatcggcg acttcaaggt ggtgggcacc ggcttccccg aggacagcgt gatcttcacc 3901 gacaagatca tccgcagcaa cgccaccgtg gagcacctgc accccatggg cgataacgtg 3961 ctggtgggca gcttcgcccg caccttcagc ctgcgcgacg gcggctacta cagcttcgtg 4021 gtggacagcc acatgcactt caagagcgcc atccacccca gcatcctgca gaacgggggc 4081 cccatgttcg ccttccgccg cgtggaggag ctgcacagca acaccgagct gggcatcgtg 4141 gagtaccagc acgccttcaa gacccccatc gccttcgcca gatcccgcgc tcagtcgtcc 4201 aattctgccg tggacggcac cgccggaccc ggctccaccg gatctcgcga gggcagagga 4261 agtcttctaa catgcggtga cgtggaggag aatcccggcc ctatgaccga gtacaagccc 4321 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg 4381 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc 4441 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc 4501 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg 4561 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag 4621 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc 4681 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc 4741 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc 4801 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa 4861 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaaatcaacc tctggattac 4921 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 4981 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 5041 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 5101 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 5161 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 5221 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 5281 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg 5341 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 5401 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg 5461 agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa tgacttacaa 5521 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca 5581 ctcccaacga agataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat 5641 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt 5701 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc 5761 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat 5821 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat 5881 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 5941 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 6001 gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 6061 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 6121 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctaga cttttgcaga 6181 gaccaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat 6241 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 6301 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 6361 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 6421 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 6481 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 6541 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 6601 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 6661 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 6721 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 6781 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 6841 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 6901 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 6961 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 7021 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 7081 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 7141 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 7201 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 7261 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 7321 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 7381 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 7441 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 7501 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 7561 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 7621 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 7681 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 7741 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 7801 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 7861 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 7921 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 7981 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 8041 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 8101 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 8161 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 8221 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 8281 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 8341 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 8401 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 8461 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 8521 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 8581 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 8641 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 8701 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 8761 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 8821 gctg //