LOCUS V028961 9133 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V028961 VERSION V028961 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..3159 /label="Eapp(NM_025456)" /note="Eapp(NM_025456)" /gene="Eapp" CDS 3166..3231 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3275..3486 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3499..3767 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3796..3805 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3826..4488 /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 4558..4611 /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 4612..5211 /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 5212..5800 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5874..6107 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6179..6300 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6340..6475 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6508..6524) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6532..6548 /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(6556..6586) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6601..6622 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6910..7498) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7669..8529) /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(8530..8634) /gene="bla" /label="AmpR promoter" primer_bind 9108..9124 /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 accgactcca ggatgactat 2341 gacccgtacg cggtggaaga gccgagcgac gaggagccgg ctctgagcag ttctgaagat 2401 gaactggatg tgcttttaca tggaacccca gaccaaaagc gaaaactcat ccgggaatgt 2461 cttactggag aaagtgagtc atcaagtgaa gatgaatttg aaaaagaaat ggaggctgaa 2521 ctaaactcca ccatgaagac aatggaggac cagttatcct cactgggaac agggtcttct 2581 tcaggagttg ccaaagtggg aggagtcaca gaaaagttct atgatgagat atattttgat 2641 tctgattctg aggatgagga caaaacagtg accaagaaga agaagaagaa acaacacagg 2701 attccaacaa acgatgagct gctgtatgat cctgagaaag ataacagaga ccaggcctgg 2761 gttgacgcca agaggagggg ttatcatgct tttggattgc aaaggccacg tcagaaacaa 2821 cagcctgttc caaacagtga tgctgttctg aattgcccag cctgtatgac gacattgtgc 2881 cttgattgcc agaggcatga atcctacaag acgcagtaca gggcaatgtt tgtgatgaat 2941 tgttctatca acagagaaga ggttctgaga tacaagaatc cagaaaacag gaggaaacgg 3001 cggagtgcta agaagatgag gtctaatcct gaagatcccg cagagagaga agcggaggaa 3061 atctatcacc ctgtcatgtg cacagagtgt tcaactgaag tggcagtcta tgacaaggat 3121 gaagtctttc actttttcaa tgttttagcg agccattctg gatccgacta caaagaccat 3181 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagcggccg 3241 cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc 3301 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg 3361 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa 3421 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga 3481 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga 3541 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga 3601 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg 3661 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct 3721 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg 3781 gcgcctacgc tagacgccac catggagagc gacgagagcg gcctgcccgc catggagatc 3841 gagtgccgca tcaccggcac cctgaacggc gtggagttcg agctggtggg cggcggagag 3901 ggcaccccca agcagggccg catgaccaac aagatgaaga gcaccaaagg cgccctgacc 3961 ttcagcccct acctgctgag ccacgtgatg ggctacggct tctaccactt cggcacctac 4021 cccagcggct acgagaaccc cttcctgcac gccatcaaca acggcggcta caccaacacc 4081 cgcatcgaga agtacgagga cggcggcgtg ctgcacgtga gcttcagcta ccgctacgag 4141 gccggccgcg tgatcggcga cttcaaggtg gtgggcaccg gcttccccga ggacagcgtg 4201 atcttcaccg acaagatcat ccgcagcaac gccaccgtgg agcacctgca ccccatgggc 4261 gataacgtgc tggtgggcag cttcgcccgc accttcagcc tgcgcgacgg cggctactac 4321 agcttcgtgg tggacagcca catgcacttc aagagcgcca tccaccccag catcctgcag 4381 aacgggggcc ccatgttcgc cttccgccgc gtggaggagc tgcacagcaa caccgagctg 4441 ggcatcgtgg agtaccagca cgccttcaag acccccatcg ccttcgccag atcccgcgct 4501 cagtcgtcca attctgccgt ggacggcacc gccggacccg gctccaccgg atctcgcgag 4561 ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgaccgag 4621 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc 4681 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc 4741 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag 4801 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa 4861 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg 4921 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg 4981 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc 5041 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc 5101 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag 5161 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aaatcaacct 5221 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 5281 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 5341 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 5401 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 5461 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 5521 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 5581 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt 5641 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 5701 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 5761 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat 5821 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg 5881 gctaattcac tcccaacgaa gataagatct gctttttgct tgtactgggt ctctctggtt 5941 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca 6001 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa 6061 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg 6121 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa 6181 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 6241 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 6301 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc 6361 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 6421 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac 6481 ttttgcagag accaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc 6541 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 6601 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 6661 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 6721 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 6781 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 6841 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 6901 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 6961 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 7021 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 7081 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 7141 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 7201 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 7261 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 7321 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 7381 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 7441 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 7501 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 7561 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 7621 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 7681 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 7741 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 7801 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 7861 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 7921 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 7981 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 8041 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 8101 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 8161 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 8221 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 8281 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 8341 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 8401 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 8461 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 8521 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 8581 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 8641 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 8701 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct 8761 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 8821 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc 8881 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg 8941 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga 9001 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc 9061 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc 9121 cagtgccaag ctg //