LOCUS V021725 8836 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V021725 VERSION V021725 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..2950 /label="FGF19(NM_005117)" /note="FGF19(NM_005117)" /gene="FGF19" promoter 2995..3206 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3219..3487 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3516..3525 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3546..4208 /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 4278..4331 /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 4332..4931 /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 4932..5519 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5593..5826 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5898..6019 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6038..6173 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6211..6227) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6235..6251 /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(6259..6289) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6304..6325 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6613..7201) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7372..8232) /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(8233..8337) /gene="bla" /label="AmpR promoter" primer_bind 8811..8827 /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 ccatgcggag cgggtgtgtg gtggtccacg tatggatcct 2341 ggccggcctc tggctggccg tggccgggcg ccccctcgcc ttctcggacg cggggcccca 2401 cgtgcactac ggctggggcg accccatccg cctgcggcac ctgtacacct ccggccccca 2461 cgggctctcc agctgcttcc tgcgcatccg tgccgacggc gtcgtggact gcgcgcgggg 2521 ccagagcgcg cacagtttgc tggagatcaa ggcagtcgct ctgcggaccg tggccatcaa 2581 gggcgtgcac agcgtgcggt acctctgcat gggcgccgac ggcaagatgc aggggctgct 2641 tcagtactcg gaggaagact gtgctttcga ggaggagatc cgcccagatg gctacaatgt 2701 gtaccgatcc gagaagcacc gcctcccggt ctccctgagc agtgccaaac agcggcagct 2761 gtacaagaac agaggctttc ttccactctc tcatttcctg cccatgctgc ccatggtccc 2821 agaggagcct gaggacctca ggggccactt ggaatctgac atgttctctt cgcccctgga 2881 gaccgacagc atggacccat ttgggcttgt caccggactg gaggccgtga ggagtcccag 2941 ctttgagaag taagcggccg cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag 3001 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaacgggtg 3061 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt 3121 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc 3181 gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc tctccttcac 3241 gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc 3301 ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga 3361 gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg ctctccacgc 3421 tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt ctgcgccgtt 3481 acagatccaa gctgtgaccg gcgcctacgc tagacgccac catggagagc gacgagagcg 3541 gcctgcccgc catggagatc gagtgccgca tcaccggcac cctgaacggc gtggagttcg 3601 agctggtggg cggcggagag ggcaccccca agcagggccg catgaccaac aagatgaaga 3661 gcaccaaagg cgccctgacc ttcagcccct acctgctgag ccacgtgatg ggctacggct 3721 tctaccactt cggcacctac cccagcggct acgagaaccc cttcctgcac gccatcaaca 3781 acggcggcta caccaacacc cgcatcgaga agtacgagga cggcggcgtg ctgcacgtga 3841 gcttcagcta ccgctacgag gccggccgcg tgatcggcga cttcaaggtg gtgggcaccg 3901 gcttccccga ggacagcgtg atcttcaccg acaagatcat ccgcagcaac gccaccgtgg 3961 agcacctgca ccccatgggc gataacgtgc tggtgggcag cttcgcccgc accttcagcc 4021 tgcgcgacgg cggctactac agcttcgtgg tggacagcca catgcacttc aagagcgcca 4081 tccaccccag catcctgcag aacgggggcc ccatgttcgc cttccgccgc gtggaggagc 4141 tgcacagcaa caccgagctg ggcatcgtgg agtaccagca cgccttcaag acccccatcg 4201 ccttcgccag atcccgcgct cagtcgtcca attctgccgt ggacggcacc gccggacccg 4261 gctccaccgg atctcgcgag ggcagaggaa gtcttctaac atgcggtgac gtggaggaga 4321 atcccggccc tatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc 4381 ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg 4441 tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg 4501 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga 4561 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg 4621 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc 4681 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca 4741 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc 4801 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca 4861 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc 4921 ccggtgcctg aaatcaacct ctggattaca aaatttgtga aagattgact ggtattctta 4981 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta 5041 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt 5101 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg 5161 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt 5221 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag 5281 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc 5341 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc 5401 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc 5461 ttccgcgtct ccgccttcgc cctcagacga gtcggatctc cctttggccg cctccccgcc 5521 tggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag 5581 aaaagggggg actggaaggg ctaattcact cccaacgaaa ataagatctg ctttttgctt 5641 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga 5701 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc 5761 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct 5821 ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa 5881 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata 5941 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 6001 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccag 6061 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc 6121 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagactt 6181 ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 6241 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 6301 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 6361 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 6421 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 6481 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 6541 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 6601 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 6661 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 6721 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 6781 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 6841 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 6901 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 6961 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 7021 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct 7081 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 7141 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 7201 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 7261 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 7321 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 7381 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 7441 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 7501 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 7561 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 7621 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 7681 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 7741 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 7801 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 7861 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 7921 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 7981 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 8041 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 8101 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 8161 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 8221 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 8281 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 8341 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta 8401 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa 8461 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag 8521 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta 8581 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag 8641 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg 8701 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc 8761 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac 8821 ggccagtgcc aagctg //