LOCUS V028895 8731 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V028895 VERSION V028895 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..2757 /label="Hbb-bs(NM_001201391)" /note="Hbb-bs(NM_001201391)" /gene="Hbb-bs" CDS 2764..2829 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 2873..3084 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3097..3365 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3394..3403 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3424..4086 /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 4156..4209 /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 4210..4809 /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 4810..5398 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5472..5705 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5777..5898 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5938..6073 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6106..6122) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6130..6146 /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(6154..6184) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6199..6220 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6508..7096) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7267..8127) /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(8128..8232) /gene="bla" /label="AmpR promoter" primer_bind 8706..8722 /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 tgcacctgac tgatgctgag 2341 aaggctgctg tctctggcct gtggggaaag gtgaacgccg atgaagttgg tggtgaggcc 2401 ctgggcaggc tgctggttgt ctacccttgg acccagcggt actttgatag ctttggagac 2461 ctatcctctg cctctgctat catgggtaat gccaaagtga aggcccatgg caagaaagtg 2521 ataactgcct ttaacgatgg cctgaatcac ttggacagcc tcaagggcac ctttgccagc 2581 ctcagtgagc tccactgtga caagctgcat gtggatcctg agaacttcag gctcctgggc 2641 aatatgatcg tgattgtgct gggccaccac ctgggcaagg atttcacccc cgctgcacag 2701 gctgccttcc agaaggtggt ggctggagtg gccgctgccc tggctcacaa gtaccacgga 2761 tccgactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac 2821 gatgacaagt agcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag 2881 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc 2941 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt 3001 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc 3061 aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc 3121 gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc 3181 gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga 3241 ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt 3301 tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac 3361 agatccaagc tgtgaccggc gcctacgcta gacgccacca tggagagcga cgagagcggc 3421 ctgcccgcca tggagatcga gtgccgcatc accggcaccc tgaacggcgt ggagttcgag 3481 ctggtgggcg gcggagaggg cacccccaag cagggccgca tgaccaacaa gatgaagagc 3541 accaaaggcg ccctgacctt cagcccctac ctgctgagcc acgtgatggg ctacggcttc 3601 taccacttcg gcacctaccc cagcggctac gagaacccct tcctgcacgc catcaacaac 3661 ggcggctaca ccaacacccg catcgagaag tacgaggacg gcggcgtgct gcacgtgagc 3721 ttcagctacc gctacgaggc cggccgcgtg atcggcgact tcaaggtggt gggcaccggc 3781 ttccccgagg acagcgtgat cttcaccgac aagatcatcc gcagcaacgc caccgtggag 3841 cacctgcacc ccatgggcga taacgtgctg gtgggcagct tcgcccgcac cttcagcctg 3901 cgcgacggcg gctactacag cttcgtggtg gacagccaca tgcacttcaa gagcgccatc 3961 caccccagca tcctgcagaa cgggggcccc atgttcgcct tccgccgcgt ggaggagctg 4021 cacagcaaca ccgagctggg catcgtggag taccagcacg ccttcaagac ccccatcgcc 4081 ttcgccagat cccgcgctca gtcgtccaat tctgccgtgg acggcaccgc cggacccggc 4141 tccaccggat ctcgcgaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat 4201 cccggcccta tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc 4261 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc 4321 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc 4381 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc 4441 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag 4501 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg 4561 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag 4621 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc 4681 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc 4741 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc 4801 ggtgcctgaa atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac 4861 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt 4921 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat 4981 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca 5041 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc 5101 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg 5161 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct 5221 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct 5281 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt 5341 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct 5401 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga 5461 aaagggggga ctggaagggc taattcactc ccaacgaaga taagatctgc tttttgcttg 5521 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa 5581 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct 5641 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc 5701 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat 5761 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag 5821 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 5881 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc 5941 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 6001 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc 6061 ttttttggag gcctagactt ttgcagagac caaattcgta atcatgtcat agctgtttcc 6121 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 6181 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 6241 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 6301 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 6361 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 6421 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 6481 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 6541 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 6601 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 6661 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 6721 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 6781 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 6841 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 6901 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 6961 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 7021 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 7081 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 7141 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 7201 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 7261 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 7321 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 7381 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 7441 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 7501 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 7561 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 7621 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 7681 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 7741 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 7801 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 7861 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 7921 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 7981 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 8041 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 8101 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 8161 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 8221 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat 8281 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg 8341 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta 8401 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg 8461 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg 8521 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag 8581 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc 8641 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg 8701 acgttgtaaa acgacggcca gtgccaagct g //