LOCUS V038133 8766 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V038133 VERSION V038133 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 2303..2851 /label="Ube2m(NM_001108471)" /note="Ube2m(NM_001108471)" /gene="Ube2m" CDS 2858..2881 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" promoter 2925..3136 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3149..3417 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3446..3455 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3476..4138 /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 4208..4261 /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 4262..4861 /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 4862..5449 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5523..5756 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5828..5949 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5968..6103 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6141..6157) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6165..6181 /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(6189..6219) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6234..6255 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6543..7131) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7302..8162) /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(8163..8267) /gene="bla" /label="AmpR promoter" primer_bind 8741..8757 /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 gaattcgcca ccatgatcaa gctgttctcg ctgaagcagc agaagaagga 2341 ggaggagtcg gccggcggca ccaagggcag cagcaagaag gcgtcggcgg cgcagctgcg 2401 gattcagaag gacattaacg agctgaacct gcccaagacg tgtgacatca gcttctcaga 2461 cccagacgac ctcctcaact tcaaactggt tatctgtcct gatgaaggct tctacaagag 2521 cggcaagttt gtgttcagct ttaaggtggg acagggttac ccacatgacc ctcccaaggt 2581 gaagtgtgaa acaatggttt atcatcccaa cattgacctc gagggcaacg tctgcctcaa 2641 catcctcaga gaggactgga agccagtcct tacgataaac tccataattt atggcctgca 2701 gtatctcttc ttggagccca accccgagga cccactgaac aaggaggctg ccgaggtcct 2761 gcagaacaat cggcggctgt ttgaacaaaa cgtgcagcgc tccatgagag gtggttacat 2821 cgggtccacc tacttcgagc gctgcctgaa aggatccgat tacaaggacg acgatgacaa 2881 gtagcggccg cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc 2941 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag 3001 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg 3061 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt 3121 tgccgccaga acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg 3181 ccctacctga ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg 3241 tgcctcctga actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct 3301 ttgtccggcg ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac 3361 cctgcttgct caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa 3421 gctgtgaccg gcgcctacgc tagacgccac catggagagc gacgagagcg gcctgcccgc 3481 catggagatc gagtgccgca tcaccggcac cctgaacggc gtggagttcg agctggtggg 3541 cggcggagag ggcaccccca agcagggccg catgaccaac aagatgaaga gcaccaaagg 3601 cgccctgacc ttcagcccct acctgctgag ccacgtgatg ggctacggct tctaccactt 3661 cggcacctac cccagcggct acgagaaccc cttcctgcac gccatcaaca acggcggcta 3721 caccaacacc cgcatcgaga agtacgagga cggcggcgtg ctgcacgtga gcttcagcta 3781 ccgctacgag gccggccgcg tgatcggcga cttcaaggtg gtgggcaccg gcttccccga 3841 ggacagcgtg atcttcaccg acaagatcat ccgcagcaac gccaccgtgg agcacctgca 3901 ccccatgggc gataacgtgc tggtgggcag cttcgcccgc accttcagcc tgcgcgacgg 3961 cggctactac agcttcgtgg tggacagcca catgcacttc aagagcgcca tccaccccag 4021 catcctgcag aacgggggcc ccatgttcgc cttccgccgc gtggaggagc tgcacagcaa 4081 caccgagctg ggcatcgtgg agtaccagca cgccttcaag acccccatcg ccttcgccag 4141 atcccgcgct cagtcgtcca attctgccgt ggacggcacc gccggacccg gctccaccgg 4201 atctcgcgag ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc 4261 tatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt 4321 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga 4381 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga 4441 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga 4501 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg 4561 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga 4621 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg 4681 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct 4741 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc 4801 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg 4861 aaatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 4921 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 4981 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 5041 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 5101 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 5161 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 5221 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct 5281 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 5341 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 5401 ccgccttcgc cctcagacga gtcggatctc cctttggccg cctccccgcc tggtaccttt 5461 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg 5521 actggaaggg ctaattcact cccaacgaaa ataagatctg ctttttgctt gtactgggtc 5581 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct 5641 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga 5701 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag 5761 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag 5821 tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa 5881 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa 5941 tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccag ttccgcccat 6001 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc 6061 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagactt ttgcagagac 6121 ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca 6181 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg 6241 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg 6301 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc 6361 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 6421 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 6481 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 6541 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 6601 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 6661 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 6721 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 6781 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 6841 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 6901 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 6961 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt 7021 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 7081 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 7141 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 7201 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 7261 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 7321 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 7381 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 7441 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 7501 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 7561 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 7621 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 7681 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 7741 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 7801 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 7861 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 7921 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 7981 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 8041 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 8101 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 8161 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 8221 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 8281 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg 8341 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac 8401 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc 8461 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca 8521 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg 8581 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga 8641 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga 8701 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc 8761 aagctg //