LOCUS V029157 9553 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029157 VERSION V029157 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..3579 /label="Syt1(NM_001252341)" /note="Syt1(NM_001252341)" /gene="Syt1" CDS 3586..3651 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3695..3906 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3919..4187 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4216..4225 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4246..4908 /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 4978..5031 /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 5032..5631 /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 5632..6220 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6294..6527 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6599..6720 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6760..6895 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6928..6944) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6952..6968 /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(6976..7006) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7021..7042 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7330..7918) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8089..8949) /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(8950..9054) /gene="bla" /label="AmpR promoter" primer_bind 9528..9544 /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 tgagtgccag tcgtcctgag 2341 gccctggctg cccctgtcac cactgttgcg acccttgtcc cacacaacgc cactgagcca 2401 gccagtcctg gggaagggaa ggaagatgcc ttttccaagc tgaagcagaa gtttatgaat 2461 gaactgcata aaatcccatt gccaccgtgg gccttaattg ccatagccat agttgcggtc 2521 cttctagtcg tgacctgctg cttctgtgtc tgtaagaaat gtttgttcaa aaagaaaaac 2581 aagaagaagg gaaaggaaaa gggagggaag aacgccatta acatgaaaga cgtgaaagac 2641 ttagggaaga ccatgaagga tcaggccctt aaggatgacg atgctgaaac tggactgact 2701 gatggagaag aaaaggagga gcccaaggaa gaggagaaac tgggaaagct tcaatattca 2761 ctggactatg acttccagaa taaccagctg ctggtgggaa tcatccaggc tgctgaactg 2821 cccgccctgg acatgggagg cacatctgat ccatacgtca aagtcttcct gctgcccgac 2881 aaaaagaaga agtttgagac aaaagtccac cggaaaaccc tcaatccagt cttcaatgaa 2941 cagtttactt tcaaggtgcc atactcggaa ttaggtggca agacactggt gatggctgtg 3001 tatgattttg accgcttctc caagcacgac atcattggag agttcaaagt tcctatgaac 3061 accgtggatt ttggccacgt caccgaggag tggcgcgatc tccagagtgc tgagaaagaa 3121 gagcaagaga aactgggtga catctgcttc tccctccgct acgtccctac tgccggcaag 3181 ctgactgttg tcattctgga agccaagaac ctgaagaaga tggatgtggg tggcttatct 3241 gatccctatg taaagattca cctgatgcag aacggcaaga gactgaagaa gaaaaagaca 3301 acgattaaga agaacacact taacccctac tacaatgagt ccttcagctt tgaagttccg 3361 ttcgagcaaa tccagaaagt gcaagtggtg gtaactgttt tggactatga caagattggc 3421 aagaacgacg ccatcggcaa agtctttgtg ggctacaaca gcaccggcgc agagctgcga 3481 cactggtcag acatgctggc caacccccgg cgacccatcg cccagtggca cactctgcag 3541 gtagaggagg aggttgatgc catgctggct gtcaagaagg gatccgacta caaagaccat 3601 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagcggccg 3661 cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc 3721 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg 3781 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa 3841 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga 3901 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga 3961 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga 4021 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg 4081 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct 4141 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg 4201 gcgcctacgc tagacgccac catggagagc gacgagagcg gcctgcccgc catggagatc 4261 gagtgccgca tcaccggcac cctgaacggc gtggagttcg agctggtggg cggcggagag 4321 ggcaccccca agcagggccg catgaccaac aagatgaaga gcaccaaagg cgccctgacc 4381 ttcagcccct acctgctgag ccacgtgatg ggctacggct tctaccactt cggcacctac 4441 cccagcggct acgagaaccc cttcctgcac gccatcaaca acggcggcta caccaacacc 4501 cgcatcgaga agtacgagga cggcggcgtg ctgcacgtga gcttcagcta ccgctacgag 4561 gccggccgcg tgatcggcga cttcaaggtg gtgggcaccg gcttccccga ggacagcgtg 4621 atcttcaccg acaagatcat ccgcagcaac gccaccgtgg agcacctgca ccccatgggc 4681 gataacgtgc tggtgggcag cttcgcccgc accttcagcc tgcgcgacgg cggctactac 4741 agcttcgtgg tggacagcca catgcacttc aagagcgcca tccaccccag catcctgcag 4801 aacgggggcc ccatgttcgc cttccgccgc gtggaggagc tgcacagcaa caccgagctg 4861 ggcatcgtgg agtaccagca cgccttcaag acccccatcg ccttcgccag atcccgcgct 4921 cagtcgtcca attctgccgt ggacggcacc gccggacccg gctccaccgg atctcgcgag 4981 ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgaccgag 5041 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc 5101 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc 5161 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag 5221 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa 5281 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg 5341 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg 5401 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc 5461 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc 5521 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag 5581 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aaatcaacct 5641 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 5701 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 5761 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 5821 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 5881 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 5941 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 6001 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt 6061 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 6121 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 6181 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat 6241 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg 6301 gctaattcac tcccaacgaa gataagatct gctttttgct tgtactgggt ctctctggtt 6361 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca 6421 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa 6481 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg 6541 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa 6601 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 6661 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 6721 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc 6781 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 6841 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac 6901 ttttgcagag accaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc 6961 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 7021 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 7081 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 7141 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 7201 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 7261 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 7321 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 7381 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 7441 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 7501 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 7561 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 7621 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 7681 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 7741 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 7801 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 7861 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 7921 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 7981 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 8041 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 8101 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 8161 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 8221 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 8281 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 8341 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 8401 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 8461 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 8521 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 8581 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 8641 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 8701 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 8761 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 8821 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 8881 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 8941 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 9001 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 9061 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 9121 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct 9181 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 9241 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc 9301 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg 9361 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga 9421 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc 9481 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc 9541 cagtgccaag ctg //