LOCUS V045574 9436 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045574 VERSION V045574 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..3462 /label="S1PR1(NM_001320730)" /note="S1PR1(NM_001320730)" /gene="S1PR1" CDS 3469..3534 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3578..3789 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3802..4070 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4099..4108 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4129..4791 /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 4861..4914 /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 4915..5514 /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 5515..6103 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6177..6410 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6482..6603 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6643..6778 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6811..6827) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6835..6851 /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(6859..6889) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6904..6925 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7213..7801) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7972..8832) /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(8833..8937) /gene="bla" /label="AmpR promoter" primer_bind 9411..9427 /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 ggcccaccag cgtcccgctg 2341 gtcaaggccc accgcagctc ggtctctgac tacgtcaact atgatatcat cgtccggcat 2401 tacaactaca cgggaaagct gaatatcagc gcggacaagg agaacagcat taaactgacc 2461 tcggtggtgt tcattctcat ctgctgcttt atcatcctgg agaacatctt tgtcttgctg 2521 accatttgga aaaccaagaa attccaccga cccatgtact attttattgg caatctggcc 2581 ctctcagacc tgttggcagg agtagcctac acagctaacc tgctcttgtc tggggccacc 2641 acctacaagc tcactcccgc ccagtggttt ctgcgggaag ggagtatgtt tgtggccctg 2701 tcagcctccg tgttcagtct cctcgccatc gccattgagc gctatatcac aatgctgaaa 2761 atgaaactcc acaacgggag caataacttc cgcctcttcc tgctaatcag cgcctgctgg 2821 gtcatctccc tcatcctggg tggcctgcct atcatgggct ggaactgcat cagtgcgctg 2881 tccagctgct ccaccgtgct gccgctctac cacaagcact atatcctctt ctgcaccacg 2941 gtcttcactc tgcttctgct ctccatcgtc attctgtact gcagaatcta ctccttggtc 3001 aggactcgga gccgccgcct gacgttccgc aagaacattt ccaaggccag ccgcagctct 3061 gagaagtcgc tggcgctgct caagaccgta attatcgtcc tgagcgtctt catcgcctgc 3121 tgggcaccgc tcttcatcct gctcctgctg gatgtgggct gcaaggtgaa gacctgtgac 3181 atcctcttca gagcggagta cttcctggtg ttagctgtgc tcaactccgg caccaacccc 3241 atcatttaca ctctgaccaa caaggagatg cgtcgggcct tcatccggat catgtcctgc 3301 tgcaagtgcc cgagcggaga ctctgctggc aaattcaagc gacccatcat cgccggcatg 3361 gaattcagcc gcagcaaatc ggacaattcc tcccaccccc agaaagacga aggggacaac 3421 ccagagacca ttatgtcttc tggaaacgtc aactcttctt ccggatccga ctacaaagac 3481 catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga caagtagcgg 3541 ccgcgaagga tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca 3601 gtccccgaga agttgggggg aggggtcggc aattgaacgg gtgcctagag aaggtggcgc 3661 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga 3721 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc 3781 agaacacagc tgaagcttcg aggggctcgc atctctcctt cacgcgcccg ccgccctacc 3841 tgaggccgcc atccacgccg gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc 3901 tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt cgagaccggg cctttgtccg 3961 gcgctccctt ggagcctacc tagactcagc cggctctcca cgctttgcct gaccctgctt 4021 gctcaactct acgtctttgt ttcgttttct gttctgcgcc gttacagatc caagctgtga 4081 ccggcgccta cgctagacgc caccatggag agcgacgaga gcggcctgcc cgccatggag 4141 atcgagtgcc gcatcaccgg caccctgaac ggcgtggagt tcgagctggt gggcggcgga 4201 gagggcaccc ccaagcaggg ccgcatgacc aacaagatga agagcaccaa aggcgccctg 4261 accttcagcc cctacctgct gagccacgtg atgggctacg gcttctacca cttcggcacc 4321 taccccagcg gctacgagaa ccccttcctg cacgccatca acaacggcgg ctacaccaac 4381 acccgcatcg agaagtacga ggacggcggc gtgctgcacg tgagcttcag ctaccgctac 4441 gaggccggcc gcgtgatcgg cgacttcaag gtggtgggca ccggcttccc cgaggacagc 4501 gtgatcttca ccgacaagat catccgcagc aacgccaccg tggagcacct gcaccccatg 4561 ggcgataacg tgctggtggg cagcttcgcc cgcaccttca gcctgcgcga cggcggctac 4621 tacagcttcg tggtggacag ccacatgcac ttcaagagcg ccatccaccc cagcatcctg 4681 cagaacgggg gccccatgtt cgccttccgc cgcgtggagg agctgcacag caacaccgag 4741 ctgggcatcg tggagtacca gcacgccttc aagaccccca tcgccttcgc cagatcccgc 4801 gctcagtcgt ccaattctgc cgtggacggc accgccggac ccggctccac cggatctcgc 4861 gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatcccgg ccctatgacc 4921 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc 4981 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac 5041 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc 5101 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc 5161 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg 5221 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg 5281 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc 5341 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc 5401 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc 5461 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgaaatcaa 5521 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt 5581 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct 5641 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc 5701 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg 5761 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc 5821 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc 5881 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt 5941 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca 6001 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt 6061 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggtac ctttaagacc 6121 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga 6181 agggctaatt cactcccaac gaagataaga tctgcttttt gcttgtactg ggtctctctg 6241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc 6301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg 6361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc 6421 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag 6481 gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 6541 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 6601 ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg 6661 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 6721 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 6781 gacttttgca gagaccaaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta 6841 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 6901 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 6961 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 7021 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 7081 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 7141 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 7201 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 7261 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 7321 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 7381 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 7441 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 7501 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 7561 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 7621 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct 7681 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 7741 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 7801 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 7861 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 7921 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 7981 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 8041 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 8101 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 8161 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 8221 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 8281 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 8341 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 8401 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 8461 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 8521 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 8581 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 8641 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 8701 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 8761 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 8821 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 8881 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 8941 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta 9001 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa 9061 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag 9121 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta 9181 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag 9241 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg 9301 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc 9361 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac 9421 ggccagtgcc aagctg //