LOCUS V036294 9103 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V036294 VERSION V036294 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..3129 /label="Cela2a(NM_007919)" /note="Cela2a(NM_007919)" /gene="Cela2a" CDS 3136..3201 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3245..3456 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3469..3737 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3766..3775 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3796..4458 /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 4528..4581 /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 4582..5181 /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 5182..5770 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5844..6077 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6149..6270 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6310..6445 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6478..6494) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6502..6518 /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(6526..6556) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6571..6592 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6880..7468) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7639..8499) /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(8500..8604) /gene="bla" /label="AmpR promoter" primer_bind 9078..9094 /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 tcaggacact gctgctatct 2341 gccttggtgg ctggagccct cagctgtggg taccccactt atgaggtgga ggatgatgtg 2401 agcagggtag ttgggggtca agaggccaca cccaacacct ggccctggca ggtctccctg 2461 caggtccttt cctccggaag gtggcgccac aactgcggag gctccctggt ggccaacaac 2521 tgggttctga cagctgccca ttgcctcagc aactatcaga cctaccgagt gctgctgggc 2581 gcacacagcc tctccaaccc cggagctggc tctgctgctg ttcaagtctc taagcttgtg 2641 gtccaccaga ggtggaactc ccaaaacgtc ggcaatggct atgacattgc cttaatcaaa 2701 ctggccagcc cagtgaccct gagcaagaac atccagacag cttgcctccc acccgctggc 2761 accattctcc cgagaaacta tgtctgctat gtcacaggct ggggcctgct gcagaccaat 2821 gggaacagtc ctgacaccct gaggcagggc cgcctgctgg ttgtggacta tgccacctgc 2881 tccagcgcta gctggtgggg aagctctgtg aagtccagca tggtgtgcgc tggtggcgac 2941 ggcgtgacct ccagctgcaa tggggactct ggcggaccac tgaattgccg ggcatctaat 3001 ggccagtggc aggtgcatgg catcgtgagc ttcggctcct ctctgggctg caactacccc 3061 cgcaagccat ccgtcttcac cagggtctcc aactacattg actggatcaa ctcggtgatg 3121 gcaaggaacg gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac 3181 tacaaggatg acgatgacaa gtagcggccg cgaaggatct gcgatcgctc cggtgcccgt 3241 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat 3301 tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg 3361 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac 3421 gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc 3481 tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt 3541 ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag 3601 ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg 3661 ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt 3721 ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagacgccac catggagagc 3781 gacgagagcg gcctgcccgc catggagatc gagtgccgca tcaccggcac cctgaacggc 3841 gtggagttcg agctggtggg cggcggagag ggcaccccca agcagggccg catgaccaac 3901 aagatgaaga gcaccaaagg cgccctgacc ttcagcccct acctgctgag ccacgtgatg 3961 ggctacggct tctaccactt cggcacctac cccagcggct acgagaaccc cttcctgcac 4021 gccatcaaca acggcggcta caccaacacc cgcatcgaga agtacgagga cggcggcgtg 4081 ctgcacgtga gcttcagcta ccgctacgag gccggccgcg tgatcggcga cttcaaggtg 4141 gtgggcaccg gcttccccga ggacagcgtg atcttcaccg acaagatcat ccgcagcaac 4201 gccaccgtgg agcacctgca ccccatgggc gataacgtgc tggtgggcag cttcgcccgc 4261 accttcagcc tgcgcgacgg cggctactac agcttcgtgg tggacagcca catgcacttc 4321 aagagcgcca tccaccccag catcctgcag aacgggggcc ccatgttcgc cttccgccgc 4381 gtggaggagc tgcacagcaa caccgagctg ggcatcgtgg agtaccagca cgccttcaag 4441 acccccatcg ccttcgccag atcccgcgct cagtcgtcca attctgccgt ggacggcacc 4501 gccggacccg gctccaccgg atctcgcgag ggcagaggaa gtcttctaac atgcggtgac 4561 gtggaggaga atcccggccc tatgaccgag tacaagccca cggtgcgcct cgccacccgc 4621 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg 4681 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc 4741 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg 4801 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg 4861 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg 4921 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac 4981 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc 5041 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg 5101 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg 5161 acccgcaagc ccggtgcctg aaatcaacct ctggattaca aaatttgtga aagattgact 5221 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg 5281 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg 5341 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg 5401 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg 5461 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc 5521 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca 5581 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc 5641 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct 5701 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc 5761 gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca 5821 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gataagatct 5881 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg 5941 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt 6001 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt 6061 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc 6121 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa 6181 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 6241 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta 6301 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga 6361 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag 6421 tagtgaggag gcttttttgg aggcctagac ttttgcagag accaaattcg taatcatgtc 6481 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg 6541 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt 6601 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 6661 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 6721 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 6781 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 6841 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 6901 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 6961 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 7021 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc 7081 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 7141 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 7201 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 7261 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 7321 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 7381 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 7441 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 7501 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 7561 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 7621 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 7681 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 7741 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 7801 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 7861 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 7921 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 7981 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 8041 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 8101 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 8161 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 8221 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 8281 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 8341 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 8401 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 8461 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 8521 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 8581 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga 8641 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct 8701 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac 8761 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt 8821 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca 8881 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca 8941 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt 9001 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt 9061 ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctg //