LOCUS V037347 10144 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V037347 VERSION V037347 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..4170 /label="Slc5a5(NM_052983)" /note="Slc5a5(NM_052983)" /gene="Slc5a5" CDS 4177..4242 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 4286..4497 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4510..4778 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4807..4816 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4837..5499 /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 5569..5622 /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 5623..6222 /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 6223..6811 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6885..7118 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 7190..7311 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7351..7486 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7519..7535) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7543..7559 /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(7567..7597) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7612..7633 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7921..8509) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8680..9540) /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(9541..9645) /gene="bla" /label="AmpR promoter" primer_bind 10119..10135 /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 agggtgcgga ggccggggcc 2341 cgggccacct tcggcgcctg ggactacggc gtgttcgcga ccatgctgct ggtgtccacg 2401 ggcatcgggc tatgggtcgg cctggcccgc ggtggccaac gcagtgccga cgacttcttt 2461 accgggggcc ggcagttggc agccgttcct gtggggctgt cgctggccgc cagtttcatg 2521 tcggctgtgc aggtgctcgg ggtccccgcc gaggcagcgc gctacgggct caagttcctg 2581 tggatgtgcg cgggtcagtt gctcaactcg ctgctcacag cgtttctctt cttgccgatc 2641 ttctaccgcc tgggccttac cagcacctac cagtacctag agctgcgctt cagccgagcg 2701 gtccggctct gcgggacgct gcagtacttg gtggccacga tgctgtatac aggcatcgtg 2761 atctacgcgc ctgcgctcat cctgaaccaa gtgaccgggt tggacatctg ggcatcgctc 2821 ctgtccacag gaatcatctg caccttgtac actaccgtgg gtggtatgaa ggccgtggtc 2881 tggacagatg tgttccaggt tgtggtaatg ctcgttggct tctgggtgat cctggcccga 2941 ggcgtcattc tcctgggggg tccccggaac atgctcagcc tcgctcagaa ccattcccgg 3001 atcaacctga tggactttga ccctgatcct cggagccggt acaccttctg gactttcata 3061 gtgggtggca cactggtgtg gctctccatg tacggtgtga accaagccca ggtacagcgc 3121 tatgtggcct gccacacaga gggaaaggcc aaactggccc tgcttgtcaa ccagctgggc 3181 ctcttcctga ttgtggccag tgcagcttgc tgtggcattg tcatgttcgt ctactacaag 3241 gactgtgacc ccctcctcac aggccgtatc tcagcccccg accagtacat gccgctgctt 3301 gtgttggaca tttttgagga tctgcccgga gtccccgggc tcttcctggc ctgtgcctac 3361 agtggcaccc tcagcactgc atccaccagc atcaacgcca tggcagctgt gactgtggaa 3421 gacctcatca agccgaggat gcctggcctg gcacctcgga agttggtttt catctctaaa 3481 gggctctcat tcatctacgg ctctgcctgc ctcactgtgg ctgctctgtc ctcactgctg 3541 ggaggtggtg tcctccaggg ttccttcact gtgatgggtg tcatcagtgg gcctctacta 3601 ggcgccttca cgcttgggat gctgctccca gcctgcaaca cgccaggcgt tctctccggg 3661 ttggcagcag gcttggctgt atccctgtgg gtggccgtag gggccacact gtatccccct 3721 ggagagcaga ccatgggggt gctgcccacc tcggctgcag gctgcaccaa cgattcggtc 3781 ctcctgggcc cacctggagc caccaacgct tccaacggga tccccagttc tggaatggac 3841 acgggccgcc ctgccctcgc tgataccttt tacgccatct cctatctcta ttacggggct 3901 ctgggcacgc tgaccaccat gctttgcggt gctctcatca gctaccttac tggtcccacc 3961 aagcgcagct ccctgggtcc cggattgctg tggtgggacc ttgctcgaca gacagcgtct 4021 gtggccccaa aggaagacac tgccaccctg gaggagagcc tggtgaaggg accggaagac 4081 atccctgctg tgaccaagaa gccccctggc ctcaagccag gcgccgagac ccaccccctg 4141 tatctggggc acgatgtgga gaccaacctc ggatccgact acaaagacca tgacggtgat 4201 tataaagatc atgacatcga ctacaaggat gacgatgaca agtagcggcc gcgaaggatc 4261 tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag 4321 ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg 4381 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata 4441 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg 4501 aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat 4561 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc 4621 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg 4681 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac 4741 gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg 4801 ctagacgcca ccatggagag cgacgagagc ggcctgcccg ccatggagat cgagtgccgc 4861 atcaccggca ccctgaacgg cgtggagttc gagctggtgg gcggcggaga gggcaccccc 4921 aagcagggcc gcatgaccaa caagatgaag agcaccaaag gcgccctgac cttcagcccc 4981 tacctgctga gccacgtgat gggctacggc ttctaccact tcggcaccta ccccagcggc 5041 tacgagaacc ccttcctgca cgccatcaac aacggcggct acaccaacac ccgcatcgag 5101 aagtacgagg acggcggcgt gctgcacgtg agcttcagct accgctacga ggccggccgc 5161 gtgatcggcg acttcaaggt ggtgggcacc ggcttccccg aggacagcgt gatcttcacc 5221 gacaagatca tccgcagcaa cgccaccgtg gagcacctgc accccatggg cgataacgtg 5281 ctggtgggca gcttcgcccg caccttcagc ctgcgcgacg gcggctacta cagcttcgtg 5341 gtggacagcc acatgcactt caagagcgcc atccacccca gcatcctgca gaacgggggc 5401 cccatgttcg ccttccgccg cgtggaggag ctgcacagca acaccgagct gggcatcgtg 5461 gagtaccagc acgccttcaa gacccccatc gccttcgcca gatcccgcgc tcagtcgtcc 5521 aattctgccg tggacggcac cgccggaccc ggctccaccg gatctcgcga gggcagagga 5581 agtcttctaa catgcggtga cgtggaggag aatcccggcc ctatgaccga gtacaagccc 5641 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg 5701 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc 5761 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc 5821 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg 5881 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag 5941 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc 6001 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc 6061 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc 6121 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa 6181 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaaatcaacc tctggattac 6241 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 6301 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 6361 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 6421 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 6481 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 6541 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 6601 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg 6661 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 6721 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg 6781 agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa tgacttacaa 6841 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca 6901 ctcccaacga agataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat 6961 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt 7021 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc 7081 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat 7141 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat 7201 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 7261 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 7321 gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 7381 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 7441 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctaga cttttgcaga 7501 gaccaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat 7561 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 7621 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 7681 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 7741 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 7801 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 7861 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 7921 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 7981 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 8041 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 8101 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 8161 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 8221 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 8281 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 8341 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 8401 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 8461 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 8521 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 8581 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 8641 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 8701 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 8761 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 8821 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 8881 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 8941 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 9001 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 9061 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 9121 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 9181 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 9241 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 9301 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 9361 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 9421 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 9481 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 9541 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 9601 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 9661 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 9721 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 9781 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 9841 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 9901 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 9961 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 10021 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 10081 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 10141 gctg //