LOCUS V033124 8764 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V033124 VERSION V033124 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..2790 /label="SRGN(NM_002727)" /note="SRGN(NM_002727)" /gene="SRGN" CDS 2797..2862 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 2906..3117 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3130..3398 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3427..3436 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3457..4119 /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 4189..4242 /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 4243..4842 /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 4843..5431 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5505..5738 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5810..5931 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5971..6106 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6139..6155) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6163..6179 /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(6187..6217) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6232..6253 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6541..7129) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7300..8160) /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(8161..8265) /gene="bla" /label="AmpR promoter" primer_bind 8739..8755 /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 gccaccatga tgcagaagct actcaaatgc 2341 agtcggcttg tcctggctct tgccctcatc ctggttctgg aatcctcagt tcaaggttat 2401 cctacgcgga gagccaggta ccaatgggtg cgctgcaatc cagacagtaa ttctgcaaac 2461 tgccttgaag aaaaaggacc aatgttcgaa ctacttccag gtgaatccaa caagatcccc 2521 cgtctgagga ctgacctttt tccaaagacg agaatccagg acttgaatcg tatcttccca 2581 ctttctgagg actactctgg atcaggcttc ggctccggct ccggctctgg atcaggatct 2641 gggagtggct tcctaacgga aatggaacag gattaccaac tagtagacga aagtgatgct 2701 ttccatgaca accttaggtc tcttgacagg aatctgccct cagacagcca ggacttgggt 2761 caacatggat tagaagagga ttttatgtta ggatccgact acaaagacca tgacggtgat 2821 tataaagatc atgacatcga ctacaaggat gacgatgaca agtagcggcc gcgaaggatc 2881 tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag 2941 ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg 3001 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata 3061 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg 3121 aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat 3181 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc 3241 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg 3301 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac 3361 gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg 3421 ctagacgcca ccatggagag cgacgagagc ggcctgcccg ccatggagat cgagtgccgc 3481 atcaccggca ccctgaacgg cgtggagttc gagctggtgg gcggcggaga gggcaccccc 3541 aagcagggcc gcatgaccaa caagatgaag agcaccaaag gcgccctgac cttcagcccc 3601 tacctgctga gccacgtgat gggctacggc ttctaccact tcggcaccta ccccagcggc 3661 tacgagaacc ccttcctgca cgccatcaac aacggcggct acaccaacac ccgcatcgag 3721 aagtacgagg acggcggcgt gctgcacgtg agcttcagct accgctacga ggccggccgc 3781 gtgatcggcg acttcaaggt ggtgggcacc ggcttccccg aggacagcgt gatcttcacc 3841 gacaagatca tccgcagcaa cgccaccgtg gagcacctgc accccatggg cgataacgtg 3901 ctggtgggca gcttcgcccg caccttcagc ctgcgcgacg gcggctacta cagcttcgtg 3961 gtggacagcc acatgcactt caagagcgcc atccacccca gcatcctgca gaacgggggc 4021 cccatgttcg ccttccgccg cgtggaggag ctgcacagca acaccgagct gggcatcgtg 4081 gagtaccagc acgccttcaa gacccccatc gccttcgcca gatcccgcgc tcagtcgtcc 4141 aattctgccg tggacggcac cgccggaccc ggctccaccg gatctcgcga gggcagagga 4201 agtcttctaa catgcggtga cgtggaggag aatcccggcc ctatgaccga gtacaagccc 4261 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg 4321 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc 4381 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc 4441 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg 4501 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag 4561 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc 4621 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc 4681 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc 4741 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa 4801 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaaatcaacc tctggattac 4861 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 4921 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 4981 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 5041 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 5101 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 5161 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 5221 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg 5281 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 5341 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg 5401 agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa tgacttacaa 5461 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca 5521 ctcccaacga agataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat 5581 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt 5641 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc 5701 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat 5761 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat 5821 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 5881 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 5941 gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 6001 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 6061 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctaga cttttgcaga 6121 gaccaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat 6181 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 6241 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 6301 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 6361 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 6421 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 6481 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 6541 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 6601 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 6661 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 6721 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 6781 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 6841 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 6901 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 6961 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 7021 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 7081 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 7141 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 7201 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 7261 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 7321 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 7381 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 7441 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 7501 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 7561 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 7621 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 7681 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 7741 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 7801 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 7861 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 7921 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 7981 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 8041 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 8101 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 8161 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 8221 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 8281 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 8341 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 8401 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 8461 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 8521 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 8581 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 8641 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 8701 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 8761 gctg //