LOCUS V027445 8911 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V027445 VERSION V027445 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..2937 /label="Fgf18(NM_008005)" /note="Fgf18(NM_008005)" /gene="Fgf18" CDS 2944..3009 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3053..3264 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3277..3545 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3574..3583 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3604..4266 /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 4336..4389 /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 4390..4989 /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 4990..5578 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5652..5885 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5957..6078 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6118..6253 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6286..6302) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6310..6326 /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(6334..6364) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6379..6400 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6688..7276) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7447..8307) /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(8308..8412) /gene="bla" /label="AmpR promoter" primer_bind 8886..8902 /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 gccaccatgt attcagcgcc ctccgcctgc 2341 acttgcctgt gtttacactt tctactgctg tgcttccagg ttcaggtgtt ggcagccgag 2401 gagaatgtgg acttccgcat ccacgtggag aaccagacgc gggctcgaga tgatgtgagt 2461 cggaagcagc tgcgcttgta ccagctctat agcaggacca gtgggaagca cattcaagtc 2521 ctgggccgta ggatcagtgc ccgtggcgag gacggggaca agtatgccca gctcctagtg 2581 gagacagata ccttcgggag tcaagtccgg atcaagggca aggagacaga attctacctg 2641 tgtatgaacc gaaaaggcaa gctcgtgggg aagcctgatg gtactagcaa ggagtgcgtg 2701 ttcattgaga aggttctgga aaacaactac acggccctga tgtctgccaa gtactctggt 2761 tggtatgtgg gcttcaccaa gaaggggcgg cctcgcaagg gtcccaagac ccgcgagaac 2821 cagcaagatg tacacttcat gaagcgttac cccaagggac aggccgagct gcagaagccc 2881 ttcaaataca ccacagtcac caagcgatcc cggcggatcc gccccactca ccccggcgga 2941 tccgactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac 3001 gatgacaagt agcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag 3061 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc 3121 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt 3181 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc 3241 aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc 3301 gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc 3361 gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga 3421 ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt 3481 tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac 3541 agatccaagc tgtgaccggc gcctacgcta gacgccacca tggagagcga cgagagcggc 3601 ctgcccgcca tggagatcga gtgccgcatc accggcaccc tgaacggcgt ggagttcgag 3661 ctggtgggcg gcggagaggg cacccccaag cagggccgca tgaccaacaa gatgaagagc 3721 accaaaggcg ccctgacctt cagcccctac ctgctgagcc acgtgatggg ctacggcttc 3781 taccacttcg gcacctaccc cagcggctac gagaacccct tcctgcacgc catcaacaac 3841 ggcggctaca ccaacacccg catcgagaag tacgaggacg gcggcgtgct gcacgtgagc 3901 ttcagctacc gctacgaggc cggccgcgtg atcggcgact tcaaggtggt gggcaccggc 3961 ttccccgagg acagcgtgat cttcaccgac aagatcatcc gcagcaacgc caccgtggag 4021 cacctgcacc ccatgggcga taacgtgctg gtgggcagct tcgcccgcac cttcagcctg 4081 cgcgacggcg gctactacag cttcgtggtg gacagccaca tgcacttcaa gagcgccatc 4141 caccccagca tcctgcagaa cgggggcccc atgttcgcct tccgccgcgt ggaggagctg 4201 cacagcaaca ccgagctggg catcgtggag taccagcacg ccttcaagac ccccatcgcc 4261 ttcgccagat cccgcgctca gtcgtccaat tctgccgtgg acggcaccgc cggacccggc 4321 tccaccggat ctcgcgaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat 4381 cccggcccta tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc 4441 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc 4501 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc 4561 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc 4621 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag 4681 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg 4741 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag 4801 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc 4861 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc 4921 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc 4981 ggtgcctgaa atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac 5041 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt 5101 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat 5161 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca 5221 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc 5281 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg 5341 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct 5401 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct 5461 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt 5521 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct 5581 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga 5641 aaagggggga ctggaagggc taattcactc ccaacgaaga taagatctgc tttttgcttg 5701 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa 5761 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct 5821 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc 5881 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat 5941 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag 6001 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 6061 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc 6121 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 6181 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc 6241 ttttttggag gcctagactt ttgcagagac caaattcgta atcatgtcat agctgtttcc 6301 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 6361 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 6421 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 6481 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 6541 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 6601 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 6661 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 6721 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 6781 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 6841 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 6901 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 6961 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 7021 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 7081 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 7141 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 7201 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 7261 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 7321 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 7381 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 7441 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 7501 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 7561 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 7621 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 7681 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 7741 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 7801 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 7861 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 7921 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 7981 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 8041 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 8101 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 8161 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 8221 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 8281 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 8341 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 8401 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat 8461 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg 8521 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta 8581 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg 8641 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg 8701 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag 8761 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc 8821 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg 8881 acgttgtaaa acgacggcca gtgccaagct g //