LOCUS V028886 9853 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V028886 VERSION V028886 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..3879 /label="Creb3l2(NM_178661)" /note="Creb3l2(NM_178661)" /gene="Creb3l2" CDS 3886..3951 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3995..4206 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4219..4487 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4516..4525 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4546..5208 /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 5278..5331 /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 5332..5931 /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 5932..6520 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6594..6827 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6899..7020 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7060..7195 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7228..7244) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7252..7268 /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(7276..7306) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7321..7342 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7630..8218) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8389..9249) /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(9250..9354) /gene="bla" /label="AmpR promoter" primer_bind 9828..9844 /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 aggtgctgga gagcggggag 2341 cagagcgtcc tgcagtggga ccgcaagctg agcgagctgt cagagcccgg agagactgag 2401 gccctcatgt accacacgca cttctcggag ctcctagacg agttttccca gaacgtcctg 2461 ggtcagctcc tgagtgaccc tttcctctca gagaagagcg agtcaatgga ggtggagcca 2521 tctccaacat caccagcgcc tctcatccag gctgaacaca gctactctct gagcgaggag 2581 ccccggactc agtcaccatt tacccatgcg gctaccagcg acagcttcaa tgacgaggag 2641 gtggagagtg aaaaatggta cctgtctaca gagtttcctt cagctaccat caagacagag 2701 ccaatcacag aggagcagcc cccgggactt gtcccttctg tcactctgac catcacagcc 2761 atttccactc cttttgaaaa agaagagtcc cctctggata tgaatgctgg gggggactcc 2821 tcatgccaga cgcttattcc taagattaag ctggagccgc acgaagtgga tcagttctta 2881 aacttctccc cgaaagaagc ctccgtggat caactgcact taccaccaac accacccagt 2941 agtcacagca gtgactctga gggcagcttg agccccaacc cacgcctgca tcccttcagc 3001 ctgtctcagg cccacagccc tgccagagcc atgccccggg gcccctctgc cttgtccaca 3061 tctcctctcc tcacagctcc acataagctg cagggatcgg gccccctggt cctgacagaa 3121 gaggagaaga ggaccctggt tgccgagggc tatcccattc ccaccaagct gcctctgaca 3181 aaatctgagg agaaggccct gaagaaaatc cggagaaaga tcaagaataa gatttctgcc 3241 caagaaagca ggagaaagaa gaaagaatac atggacagcc tggagaaaaa agtggagtct 3301 tgttcaactg agaacttgga gcttcggaag aaggtggagg tgctggagaa caccaatagg 3361 actctccttc agcaacttca gaagcttcag actttggtga tggggaaggt ctctcgaacc 3421 tgcaagttag ctggcacaca gactggcacc tgcctcatgg tcgttgtgct ttgctttgct 3481 gttgcatttg gaagcttctt tcaaggctat gggccttatc cttctgccac caagatggct 3541 ctgcccagcc agcatcctct gtcagagcca tacacagcct ccgtggtgag atccaggaac 3601 ctgctaatct atgaggaaca cgctcccctg gaagagtcgt cgagcccagc ctcagccggg 3661 gagctggggg gctgggacag aggctcctct ctgctcaggg catcgtcggg gcttgaggcc 3721 ctgccagagg tggatcttcc ccatttcctt atctccaatg agacgagctt ggagaagtca 3781 gtactgttgg agcttcagca gcacctggtc agcagcaaac tggaagggaa cgaaacactc 3841 aaggttgtag agctggagag gagagtgaac gccaccttcg gatccgacta caaagaccat 3901 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagcggccg 3961 cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc 4021 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg 4081 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa 4141 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga 4201 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga 4261 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga 4321 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg 4381 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct 4441 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg 4501 gcgcctacgc tagacgccac catggagagc gacgagagcg gcctgcccgc catggagatc 4561 gagtgccgca tcaccggcac cctgaacggc gtggagttcg agctggtggg cggcggagag 4621 ggcaccccca agcagggccg catgaccaac aagatgaaga gcaccaaagg cgccctgacc 4681 ttcagcccct acctgctgag ccacgtgatg ggctacggct tctaccactt cggcacctac 4741 cccagcggct acgagaaccc cttcctgcac gccatcaaca acggcggcta caccaacacc 4801 cgcatcgaga agtacgagga cggcggcgtg ctgcacgtga gcttcagcta ccgctacgag 4861 gccggccgcg tgatcggcga cttcaaggtg gtgggcaccg gcttccccga ggacagcgtg 4921 atcttcaccg acaagatcat ccgcagcaac gccaccgtgg agcacctgca ccccatgggc 4981 gataacgtgc tggtgggcag cttcgcccgc accttcagcc tgcgcgacgg cggctactac 5041 agcttcgtgg tggacagcca catgcacttc aagagcgcca tccaccccag catcctgcag 5101 aacgggggcc ccatgttcgc cttccgccgc gtggaggagc tgcacagcaa caccgagctg 5161 ggcatcgtgg agtaccagca cgccttcaag acccccatcg ccttcgccag atcccgcgct 5221 cagtcgtcca attctgccgt ggacggcacc gccggacccg gctccaccgg atctcgcgag 5281 ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgaccgag 5341 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc 5401 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc 5461 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag 5521 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa 5581 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg 5641 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg 5701 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc 5761 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc 5821 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag 5881 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aaatcaacct 5941 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 6001 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 6061 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 6121 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 6181 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 6241 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 6301 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt 6361 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 6421 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 6481 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat 6541 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg 6601 gctaattcac tcccaacgaa gataagatct gctttttgct tgtactgggt ctctctggtt 6661 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca 6721 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa 6781 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg 6841 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa 6901 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 6961 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 7021 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc 7081 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 7141 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac 7201 ttttgcagag accaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc 7261 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 7321 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 7381 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 7441 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 7501 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 7561 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 7621 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 7681 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 7741 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 7801 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 7861 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 7921 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 7981 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 8041 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 8101 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 8161 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 8221 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 8281 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 8341 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 8401 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 8461 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 8521 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 8581 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 8641 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 8701 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 8761 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 8821 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 8881 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 8941 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 9001 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 9061 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 9121 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 9181 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 9241 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 9301 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 9361 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 9421 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct 9481 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 9541 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc 9601 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg 9661 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga 9721 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc 9781 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc 9841 cagtgccaag ctg //