LOCUS V022759 9196 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022759 VERSION V022759 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..3238 /label="SFRP1(NM_003012)" /note="SFRP1(NM_003012)" /gene="SFRP1" CDS 3245..3310 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3355..3566 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3579..3847 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 3876..3885 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 3906..4568 /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 4638..4691 /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 4692..5291 /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 5292..5879 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5953..6186 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6258..6379 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6398..6533 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6571..6587) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6595..6611 /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(6619..6649) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6664..6685 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6973..7561) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7732..8592) /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(8593..8697) /gene="bla" /label="AmpR promoter" primer_bind 9171..9187 /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 gccaccatgg gcatcgggcg cagcgagggg ggccgccgcg gggcagccct 2341 gggcgtgctg ctggcgctgg gcgcggcgct tctggccgtg ggctcggcca gcgagtacga 2401 ctacgtgagc ttccagtcgg acatcggccc gtaccagagc gggcgcttct acaccaagcc 2461 acctcagtgc gtggacatcc ccgcggacct gcggctgtgc cacaacgtgg gctacaagaa 2521 gatggtgctg cccaacctgc tggagcacga gaccatggcg gaggtgaagc agcaggccag 2581 cagctgggtg cccctgctca acaagaactg ccacgccggc acccaggtct tcctctgctc 2641 gctcttcgcg cccgtctgcc tggaccggcc catctacccg tgtcgctggc tctgcgaggc 2701 cgtgcgcgac tcgtgcgagc cggtcatgca gttcttcggc ttctactggc ccgagatgct 2761 taagtgtgac aagttccccg agggggacgt ctgcatcgcc atgacgccgc ccaatgccac 2821 cgaagcctcc aagccccaag gcacaacggt gtgtcctccc tgtgacaacg agttgaaatc 2881 tgaggccatc attgaacatc tctgtgccag cgagtttgca ctgaggatga aaataaaaga 2941 agtgaaaaaa gaaaatggcg acaagaagat tgtccccaag aagaagaagc ccctgaagtt 3001 ggggcccatc aagaagaagg acctgaagaa gcttgtgctg tacctgaaga atggggctga 3061 ctgtccctgc caccagctgg acaacctcag ccaccacttc ctcatcatgg gccgcaaggt 3121 gaagagccag tacttgctga cggccatcca caagtgggac aagaaaaaca aggagttcaa 3181 aaacttcatg aagaaaatga aaaaccatga gtgccccacc tttcagtccg tgtttaaggg 3241 atccgactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga 3301 cgatgacaag tgagcggccg cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag 3361 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaacgggtg 3421 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt 3481 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc 3541 gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc tctccttcac 3601 gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc 3661 ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga 3721 gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg ctctccacgc 3781 tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt ctgcgccgtt 3841 acagatccaa gctgtgaccg gcgcctacgc tagacgccac catggagagc gacgagagcg 3901 gcctgcccgc catggagatc gagtgccgca tcaccggcac cctgaacggc gtggagttcg 3961 agctggtggg cggcggagag ggcaccccca agcagggccg catgaccaac aagatgaaga 4021 gcaccaaagg cgccctgacc ttcagcccct acctgctgag ccacgtgatg ggctacggct 4081 tctaccactt cggcacctac cccagcggct acgagaaccc cttcctgcac gccatcaaca 4141 acggcggcta caccaacacc cgcatcgaga agtacgagga cggcggcgtg ctgcacgtga 4201 gcttcagcta ccgctacgag gccggccgcg tgatcggcga cttcaaggtg gtgggcaccg 4261 gcttccccga ggacagcgtg atcttcaccg acaagatcat ccgcagcaac gccaccgtgg 4321 agcacctgca ccccatgggc gataacgtgc tggtgggcag cttcgcccgc accttcagcc 4381 tgcgcgacgg cggctactac agcttcgtgg tggacagcca catgcacttc aagagcgcca 4441 tccaccccag catcctgcag aacgggggcc ccatgttcgc cttccgccgc gtggaggagc 4501 tgcacagcaa caccgagctg ggcatcgtgg agtaccagca cgccttcaag acccccatcg 4561 ccttcgccag atcccgcgct cagtcgtcca attctgccgt ggacggcacc gccggacccg 4621 gctccaccgg atctcgcgag ggcagaggaa gtcttctaac atgcggtgac gtggaggaga 4681 atcccggccc tatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc 4741 ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg 4801 tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg 4861 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga 4921 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg 4981 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc 5041 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca 5101 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc 5161 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca 5221 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc 5281 ccggtgcctg aaatcaacct ctggattaca aaatttgtga aagattgact ggtattctta 5341 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta 5401 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt 5461 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg 5521 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt 5581 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag 5641 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc 5701 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc 5761 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc 5821 ttccgcgtct ccgccttcgc cctcagacga gtcggatctc cctttggccg cctccccgcc 5881 tggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag 5941 aaaagggggg actggaaggg ctaattcact cccaacgaaa ataagatctg ctttttgctt 6001 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga 6061 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc 6121 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct 6181 ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa 6241 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata 6301 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 6361 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccag 6421 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc 6481 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagactt 6541 ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 6601 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 6661 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 6721 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 6781 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 6841 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 6901 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 6961 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 7021 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 7081 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 7141 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 7201 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 7261 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 7321 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 7381 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct 7441 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 7501 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 7561 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 7621 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 7681 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 7741 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 7801 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 7861 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 7921 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 7981 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 8041 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 8101 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 8161 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 8221 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 8281 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 8341 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 8401 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 8461 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 8521 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 8581 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 8641 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 8701 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta 8761 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa 8821 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag 8881 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta 8941 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag 9001 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg 9061 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc 9121 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac 9181 ggccagtgcc aagctg //