LOCUS V029035 8830 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V029035 VERSION V029035 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..2856 /label="Arxes1(NM_029541)" /note="Arxes1(NM_029541)" /gene="Arxes1" CDS 2863..2928 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 2972..3183 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3196..3464 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3493..3502 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3523..4185 /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 4255..4308 /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 4309..4908 /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 4909..5497 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5571..5804 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5876..5997 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6037..6172 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6205..6221) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6229..6245 /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(6253..6283) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6298..6319 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6607..7195) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7366..8226) /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(8227..8331) /gene="bla" /label="AmpR promoter" primer_bind 8805..8821 /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 actccctgct gtcccgggcc 2341 aactcgctgt tcgccttcac gctgagcgtc atggcggcgc tcaccttggg ctgcatcctc 2401 accaccgcct tcaaagacag gagcgcgccc gtgcgcctgc acgtctccag gatcctgctc 2461 aaaaaagtgg aagacttcac cggacccaga aagaagagcg acctgggctt catcaccttc 2521 cacatctctg cggatctgga gaagactttc gactggaacg tcaagcagct cttcctttat 2581 ctgtcggcag aatattccac caaaagcaac gctgtgaacc aggtggtcct ctgggacaag 2641 atccttctgc gaggcgagaa ccccaagctg aacctgaaag acgtcaaaag caagtatttt 2701 ttctttgacg atggacacgg gctcaaggga aacaggaatg tcactttgac tctctcctgg 2761 caagtgatac cgattgctgg gatcctgcct cttgtgactg gatctggacg cgtgtctgtc 2821 ccgtttccag attcctatga aatagccacg acttttggat ccgactacaa agaccatgac 2881 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta gcggccgcga 2941 aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc 3001 gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta 3061 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg 3121 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca 3181 cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc 3241 cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact 3301 gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc 3361 ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa 3421 ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg 3481 cctacgctag acgccaccat ggagagcgac gagagcggcc tgcccgccat ggagatcgag 3541 tgccgcatca ccggcaccct gaacggcgtg gagttcgagc tggtgggcgg cggagagggc 3601 acccccaagc agggccgcat gaccaacaag atgaagagca ccaaaggcgc cctgaccttc 3661 agcccctacc tgctgagcca cgtgatgggc tacggcttct accacttcgg cacctacccc 3721 agcggctacg agaacccctt cctgcacgcc atcaacaacg gcggctacac caacacccgc 3781 atcgagaagt acgaggacgg cggcgtgctg cacgtgagct tcagctaccg ctacgaggcc 3841 ggccgcgtga tcggcgactt caaggtggtg ggcaccggct tccccgagga cagcgtgatc 3901 ttcaccgaca agatcatccg cagcaacgcc accgtggagc acctgcaccc catgggcgat 3961 aacgtgctgg tgggcagctt cgcccgcacc ttcagcctgc gcgacggcgg ctactacagc 4021 ttcgtggtgg acagccacat gcacttcaag agcgccatcc accccagcat cctgcagaac 4081 gggggcccca tgttcgcctt ccgccgcgtg gaggagctgc acagcaacac cgagctgggc 4141 atcgtggagt accagcacgc cttcaagacc cccatcgcct tcgccagatc ccgcgctcag 4201 tcgtccaatt ctgccgtgga cggcaccgcc ggacccggct ccaccggatc tcgcgagggc 4261 agaggaagtc ttctaacatg cggtgacgtg gaggagaatc ccggccctat gaccgagtac 4321 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc 4381 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag 4441 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg 4501 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg 4561 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc 4621 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc 4681 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg 4741 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg 4801 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg 4861 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaaa tcaacctctg 4921 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta 4981 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt 5041 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc 5101 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt 5161 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg 5221 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac 5281 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc 5341 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac 5401 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct 5461 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac 5521 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct 5581 aattcactcc caacgaagat aagatctgct ttttgcttgt actgggtctc tctggttaga 5641 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata 5701 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta 5761 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca 5821 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt 5881 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 5941 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 6001 tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt 6061 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc 6121 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt 6181 tgcagagacc aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct 6241 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 6301 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 6361 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 6421 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 6481 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 6541 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 6601 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 6661 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 6721 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 6781 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 6841 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 6901 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 6961 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 7021 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 7081 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 7141 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 7201 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 7261 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 7321 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 7381 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 7441 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 7501 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 7561 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 7621 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 7681 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 7741 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 7801 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 7861 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 7921 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 7981 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 8041 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 8101 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 8161 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 8221 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 8281 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 8341 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 8401 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 8461 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 8521 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 8581 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt 8641 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg 8701 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 8761 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 8821 tgccaagctg //