LOCUS V039243 9013 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V039243 VERSION V039243 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..3039 /label="Clic1(NM_033444)" /note="Clic1(NM_033444)" /gene="Clic1" CDS 3046..3111 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3155..3366 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3379..3647 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3676..3685 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3706..4368 /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 4438..4491 /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 4492..5091 /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 5092..5680 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5754..5987 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6059..6180 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6220..6355 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6388..6404) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6412..6428 /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(6436..6466) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6481..6502 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6790..7378) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7549..8409) /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(8410..8514) /gene="bla" /label="AmpR promoter" primer_bind 8988..9004 /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 ctgaagaaca acctcaggtc 2341 gaactgttcg tgaaggctgg cagtgatggt gccaagattg ggaactgccc cttctcccag 2401 agactgttca tggtgctctg gctcaaggga gtcaccttca acgttaccac tgtggacacc 2461 aagagacgga cagagactgt acagaagctc tgccccggtg ggcagctgcc gttcttgctc 2521 tatggcaccg aagtgcacac agacaccaac aagatcgagg aattcctgga ggccatgctg 2581 tgccctccca ggtacccaaa gctggctgcc ctgaaccctg agtccaacac ctcgggactg 2641 gacatatttg caaagttttc tgcctacatc aagaactcaa acccagccct caatgacaac 2701 ctagagaagg gactcctgaa agccctgaag gttctagaca attacctgac atcccccctc 2761 ccagaagaag tggatgaaac cagcgccgaa gatgagggca tctctcagag gaagtttctg 2821 gacggcaatg agctcaccct ggctgactgc aacctgctgc caaagcttca catagtacag 2881 gtggtgtgca aaaagtacag aggcttcacc atcccagagg cattccgtgg agtgcatcgg 2941 tacttgagca acgcttatgc ccgggaagaa tttgcctcca cctgtccaga tgatgaagag 3001 atagagctag cctatgagca agtggccagg gctctcaaag gatccgacta caaagaccat 3061 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagcggccg 3121 cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc 3181 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg 3241 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa 3301 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga 3361 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga 3421 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga 3481 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg 3541 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct 3601 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg 3661 gcgcctacgc tagacgccac catggagagc gacgagagcg gcctgcccgc catggagatc 3721 gagtgccgca tcaccggcac cctgaacggc gtggagttcg agctggtggg cggcggagag 3781 ggcaccccca agcagggccg catgaccaac aagatgaaga gcaccaaagg cgccctgacc 3841 ttcagcccct acctgctgag ccacgtgatg ggctacggct tctaccactt cggcacctac 3901 cccagcggct acgagaaccc cttcctgcac gccatcaaca acggcggcta caccaacacc 3961 cgcatcgaga agtacgagga cggcggcgtg ctgcacgtga gcttcagcta ccgctacgag 4021 gccggccgcg tgatcggcga cttcaaggtg gtgggcaccg gcttccccga ggacagcgtg 4081 atcttcaccg acaagatcat ccgcagcaac gccaccgtgg agcacctgca ccccatgggc 4141 gataacgtgc tggtgggcag cttcgcccgc accttcagcc tgcgcgacgg cggctactac 4201 agcttcgtgg tggacagcca catgcacttc aagagcgcca tccaccccag catcctgcag 4261 aacgggggcc ccatgttcgc cttccgccgc gtggaggagc tgcacagcaa caccgagctg 4321 ggcatcgtgg agtaccagca cgccttcaag acccccatcg ccttcgccag atcccgcgct 4381 cagtcgtcca attctgccgt ggacggcacc gccggacccg gctccaccgg atctcgcgag 4441 ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgaccgag 4501 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc 4561 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc 4621 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag 4681 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa 4741 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg 4801 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg 4861 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc 4921 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc 4981 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag 5041 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aaatcaacct 5101 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg 5161 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc 5221 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt 5281 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc 5341 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg 5401 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact 5461 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt 5521 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg 5581 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc 5641 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat 5701 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg 5761 gctaattcac tcccaacgaa gataagatct gctttttgct tgtactgggt ctctctggtt 5821 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca 5881 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa 5941 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg 6001 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa 6061 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 6121 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 6181 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc 6241 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 6301 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac 6361 ttttgcagag accaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc 6421 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 6481 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 6541 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 6601 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 6661 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 6721 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 6781 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 6841 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 6901 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 6961 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 7021 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 7081 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 7141 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 7201 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 7261 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 7321 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 7381 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 7441 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 7501 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 7561 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 7621 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 7681 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 7741 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 7801 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 7861 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 7921 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 7981 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 8041 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 8101 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 8161 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 8221 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 8281 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 8341 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 8401 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 8461 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 8521 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 8581 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct 8641 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 8701 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc 8761 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg 8821 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga 8881 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc 8941 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc 9001 cagtgccaag ctg //