LOCUS V024765 9436 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024765 VERSION V024765 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 1798..1915 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2005..2208 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2297..3478 /label="Pa2g4(NM_011119)" /note="Pa2g4(NM_011119)" /gene="Pa2g4" CDS 3485..3550 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3595..3806 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3819..4087 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4116..4125 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4146..4808 /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 4878..4931 /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 4932..5531 /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 5532..6119 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6193..6426 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6498..6619 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6638..6773 /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 ggttaacttt 1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc 1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga 1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac 1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga 2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg 2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg 2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac 2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc 2281 tagcgaattc gccaccatgt cgggcgaaga cgagcagcag gagcaaacta tcgccgagga 2341 cctggtcgtg accaagtata agatgggggg cgacatcgcc aaccgggtgc ttcgatcttt 2401 ggtggaggct tccagctcag gtgtgtctgt gctgagcttg tgtgagaaag gtgatgccat 2461 gattatggaa gagacaggga agatcttcaa gaaggaaaag gagatgaaga aaggtattgc 2521 ctttcctacc agcatttccg taaataactg tgtgtgtcac ttctcccctt tgaagagtga 2581 ccaggactat atactcaagg aaggtgactt ggtaaaaatt gaccttgggg ttcatgtgga 2641 tggcttcatt gccaatgtgg ctcacacttt tgtaattggt gtagctcagg ggacccaggt 2701 aacagggcgg aaagcagatg tcattaaggc cgctcaccta tgtgctgaag ctgccttacg 2761 actggtcaaa cctggaaacc agaacacaca agtgactgaa gcatggaaca aggttgctca 2821 ctcatttaac tgcacaccaa tagaaggtat gctgtcacac caattgaagc agcatgtgat 2881 tgatggagag aagacgatta tccagaaccc tacagaccag cagaagaagg accatgaaaa 2941 ggcagaattt gaggtgcatg aggtttatgc tgtagatgtc ctcgtcagct caggagaagg 3001 caaggccaaa gatgcaggac agagaaccac catctacaag cgagacccct ctaaacaata 3061 tggcctgaaa atgaaaactt cacgtgcctt tttcagtgag gtggaacggc gttttgatgc 3121 catgccgttt actttaagag catttgaaga tgagaagaag gctcgaatgg gtgtggtaga 3181 gtgtgccaaa catgagttac tacagccatt taacgttctc tatgagaagg agggtgaatt 3241 tgttgcccag tttaaattta cagttctact catgcccaac ggccccatgc ggataaccag 3301 tggtcccttt gagcctgacc tgtacaagtc tgagatggag gttcaagatg cagagctgaa 3361 ggctcttctc cagagttctg caagtagaaa aacccagaaa aagaagaaaa agaaggcctc 3421 caagactgta gagaatgcca ccagtggaga aaccttagaa gagaatggag ctggggacgg 3481 atccgactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga 3541 cgatgacaag tgagcggccg cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag 3601 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaacgggtg 3661 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt 3721 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc 3781 gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc tctccttcac 3841 gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc 3901 ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga 3961 gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg ctctccacgc 4021 tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt ctgcgccgtt 4081 acagatccaa gctgtgaccg gcgcctacgc tagacgccac catggagagc gacgagagcg 4141 gcctgcccgc catggagatc gagtgccgca tcaccggcac cctgaacggc gtggagttcg 4201 agctggtggg cggcggagag ggcaccccca agcagggccg catgaccaac aagatgaaga 4261 gcaccaaagg cgccctgacc ttcagcccct acctgctgag ccacgtgatg ggctacggct 4321 tctaccactt cggcacctac cccagcggct acgagaaccc cttcctgcac gccatcaaca 4381 acggcggcta caccaacacc cgcatcgaga agtacgagga cggcggcgtg ctgcacgtga 4441 gcttcagcta ccgctacgag gccggccgcg tgatcggcga cttcaaggtg gtgggcaccg 4501 gcttccccga ggacagcgtg atcttcaccg acaagatcat ccgcagcaac gccaccgtgg 4561 agcacctgca ccccatgggc gataacgtgc tggtgggcag cttcgcccgc accttcagcc 4621 tgcgcgacgg cggctactac agcttcgtgg tggacagcca catgcacttc aagagcgcca 4681 tccaccccag catcctgcag aacgggggcc ccatgttcgc cttccgccgc gtggaggagc 4741 tgcacagcaa caccgagctg ggcatcgtgg agtaccagca cgccttcaag acccccatcg 4801 ccttcgccag atcccgcgct cagtcgtcca attctgccgt ggacggcacc gccggacccg 4861 gctccaccgg atctcgcgag ggcagaggaa gtcttctaac atgcggtgac gtggaggaga 4921 atcccggccc tatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc 4981 ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg 5041 tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg 5101 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga 5161 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg 5221 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc 5281 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca 5341 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc 5401 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca 5461 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc 5521 ccggtgcctg aaatcaacct ctggattaca aaatttgtga aagattgact ggtattctta 5581 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta 5641 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt 5701 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg 5761 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt 5821 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag 5881 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc 5941 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc 6001 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc 6061 ttccgcgtct ccgccttcgc cctcagacga gtcggatctc cctttggccg cctccccgcc 6121 tggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag 6181 aaaagggggg actggaaggg ctaattcact cccaacgaaa ataagatctg ctttttgctt 6241 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga 6301 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc 6361 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct 6421 ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa 6481 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata 6541 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 6601 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccag 6661 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc 6721 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagactt 6781 ttgcagagac ggcccaaatt cgtaatcatg 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 //