LOCUS V024718 8903 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024718 VERSION V024718 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..2946 /label="Isoc2b(NM_026158)" /note="Isoc2b(NM_026158)" /gene="Isoc2b" CDS 2953..3018 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3062..3273 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3286..3554 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3583..3592 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3613..4275 /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 4345..4398 /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 4399..4998 /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 4999..5586 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5660..5893 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5965..6086 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6105..6240 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6278..6294) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6302..6318 /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(6326..6356) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6371..6392 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6680..7268) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7439..8299) /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(8300..8404) /gene="bla" /label="AmpR promoter" primer_bind 8878..8894 /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 gagcagccaa agctagcctc 2341 ggccggatct tcccagagtc ctccatcctg ttcctatgtg acatgcagga gaaacttcga 2401 gatagaatcc tctacttccc acagatcgtg tcaaaggccg ctcggatgct taaggtagcc 2461 cagatgctag aaatccccgt cttgctgaca gagcattacc cacaaggtct gggccccaca 2521 gttcccgagc tgggtgctca gggtctgcga accatgagca aaacgtcctt cagcatggtg 2581 cccccgcttc aacaggagtt ggacaaactg ccccagctgc agtccgtgct actctgtggc 2641 attgagaccc aaggctgcat cttgcacaca gctctggacc tcctggaccg tgggctacag 2701 gttcatgtag cagtggatgc ctgttcttct caaagtgaga tgaaccggct cgtggcactg 2761 gcccgaatgc agcagagtgg cgtctttctg tccacgagcg aggtgctgat tctccagctg 2821 gtgaaggatg ctgcccaccc acagttcaag gagattcaga agatcctcaa ggagccagtc 2881 acagacatcg gactgttagg cttcttccaa ggcaagaaaa actccctgtt gcccaactcc 2941 aggaccggat ccgactacaa agaccatgac ggtgattata aagatcatga catcgactac 3001 aaggatgacg atgacaagta gcggccgcga aggatctgcg atcgctccgg tgcccgtcag 3061 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga 3121 acgggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc 3181 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt 3241 ctttttcgca acgggtttgc cgccagaaca cagctgaagc ttcgaggggc tcgcatctct 3301 ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg 3361 ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc 3421 aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc 3481 tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg 3541 cgccgttaca gatccaagct gtgaccggcg cctacgctag acgccaccat ggagagcgac 3601 gagagcggcc tgcccgccat ggagatcgag tgccgcatca ccggcaccct gaacggcgtg 3661 gagttcgagc tggtgggcgg cggagagggc acccccaagc agggccgcat gaccaacaag 3721 atgaagagca ccaaaggcgc cctgaccttc agcccctacc tgctgagcca cgtgatgggc 3781 tacggcttct accacttcgg cacctacccc agcggctacg agaacccctt cctgcacgcc 3841 atcaacaacg gcggctacac caacacccgc atcgagaagt acgaggacgg cggcgtgctg 3901 cacgtgagct tcagctaccg ctacgaggcc ggccgcgtga tcggcgactt caaggtggtg 3961 ggcaccggct tccccgagga cagcgtgatc ttcaccgaca agatcatccg cagcaacgcc 4021 accgtggagc acctgcaccc catgggcgat aacgtgctgg tgggcagctt cgcccgcacc 4081 ttcagcctgc gcgacggcgg ctactacagc ttcgtggtgg acagccacat gcacttcaag 4141 agcgccatcc accccagcat cctgcagaac gggggcccca tgttcgcctt ccgccgcgtg 4201 gaggagctgc acagcaacac cgagctgggc atcgtggagt accagcacgc cttcaagacc 4261 cccatcgcct tcgccagatc ccgcgctcag tcgtccaatt ctgccgtgga cggcaccgcc 4321 ggacccggct ccaccggatc tcgcgagggc agaggaagtc ttctaacatg cggtgacgtg 4381 gaggagaatc ccggccctat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac 4441 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc 4501 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc 4561 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg 4621 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc 4681 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg 4741 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac 4801 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc 4861 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc 4921 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc 4981 cgcaagcccg gtgcctgaaa tcaacctctg gattacaaaa tttgtgaaag attgactggt 5041 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat 5101 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg 5161 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt 5221 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact 5281 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc 5341 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg 5401 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc 5461 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg 5521 cggcctcttc cgcgtctccg ccttcgccct cagacgagtc ggatctccct ttggccgcct 5581 ccccgcctgg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt 5641 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaaaata agatctgctt 5701 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa 5761 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt 5821 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg 5881 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag 5941 aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa 6001 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt 6061 ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc 6121 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg 6181 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc 6241 tagacttttg cagagacggc ccaaattcgt aatcatggtc atagctgttt cctgtgtgaa 6301 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct 6361 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc 6421 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg 6481 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc 6541 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag 6601 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa 6661 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc 6721 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc 6781 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg 6841 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt 6901 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc 6961 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc 7021 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag 7081 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg 7141 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa 7201 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 7261 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 7321 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 7381 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 7441 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 7501 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 7561 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 7621 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 7681 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 7741 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 7801 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 7861 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 7921 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 7981 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 8041 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 8101 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 8161 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 8221 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 8281 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 8341 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 8401 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat 8461 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg 8521 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg 8581 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc 8641 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac 8701 cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca 8761 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg 8821 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta 8881 aaacgacggc cagtgccaag ctg //