LOCUS V023726 9485 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V023726 VERSION V023726 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..3528 /label="Krt33b(NM_013570)" /note="Krt33b(NM_013570)" /gene="Krt33b" CDS 3535..3600 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3644..3855 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3868..4136 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4165..4174 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4195..4857 /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 4927..4980 /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 4981..5580 /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 5581..6168 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6242..6475 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6547..6668 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6687..6822 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6860..6876) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6884..6900 /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(6908..6938) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6953..6974 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7262..7850) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8021..8881) /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(8882..8986) /gene="bla" /label="AmpR promoter" primer_bind 9460..9476 /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 gccaccatgc catacaactg ctgcctgccc 2341 gccatgagct gccgcaccag ctgctcttcc cggccctgcg tgccccccag ctgccatggc 2401 tgcaccctgc ccggggcctg caacatcccc gccaatgtgg gcaactgcaa ttggttctgt 2461 gagggctcct tcaatggcaa tgagagggag accatgcagt tcctgaatga ccgcctggcc 2521 tcctatatgg agaaggtgag gcagctggag agagagaatg cagagctgga atgtaagatc 2581 caagagagga accagcagca ggaccccctg gtgtgtcctg cctaccaggc ctacttcagg 2641 accattgagg agctgcagca gaagattctc tgtggcaagt ctgagaacgg caggctggtg 2701 gtgcagatcg acaatgccaa gctggctgct gatgacttca ggaccaagta tgagtcagag 2761 ctgtccctgc ggcagctggt ggagtcagac ctcaatggcc tgcgcaggat cctggacgaa 2821 ctgaccctct gcaagtctga tctggaggca caggtggagt ccctgaggca ggagctgctg 2881 tgtctcaagc aaaaccatga ggaggaagtc aacaccctgc gctgccagct tggagaccgc 2941 ctcaacgtag aggtggacgc tgctcccacc gtggacctga accgagtgct caacgagacc 3001 aggtgtcagt acgaggccct ggtggaaacc aaccgccggg aagtggagga atggttcacc 3061 acacagacag aggagctgaa caagcaggtg gtgtctagct cagagcagct gcagtcctgc 3121 caggcagaga tcatcgagct gagacgcaca gtcaatgccc tggagatcga gctgcaggcc 3181 cagcacagca tgagaaactc tctggagaat accctggcag agagtgaggc tcgctacagc 3241 tctcagctgt cccaggtgca gtgcctgatc accaacgtgg agtcccagct tggtgagatc 3301 cgggctgacc tggagcgtca gaaccaggag taccaggtgt tgctggatgt caaggcccgg 3361 ctggagtgtg agatcaacac gtataggggc ctgctggaga gcgaggactg caagctgccc 3421 tgcaacccct gtgccaccac caatgcctgt gacaagccca ttggaccctg tgtcacaaac 3481 ccttgtgtca cacgctcccg atgtggaccc tgcaattcct ttggctgcgg atccgactac 3541 aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag 3601 tagcggccgc gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg 3661 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg 3721 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt 3781 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt 3841 gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc 3901 cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt 3961 gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt 4021 tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc 4081 ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag 4141 ctgtgaccgg cgcctacgct agacgccacc atggagagcg acgagagcgg cctgcccgcc 4201 atggagatcg agtgccgcat caccggcacc ctgaacggcg tggagttcga gctggtgggc 4261 ggcggagagg gcacccccaa gcagggccgc atgaccaaca agatgaagag caccaaaggc 4321 gccctgacct tcagccccta cctgctgagc cacgtgatgg gctacggctt ctaccacttc 4381 ggcacctacc ccagcggcta cgagaacccc ttcctgcacg ccatcaacaa cggcggctac 4441 accaacaccc gcatcgagaa gtacgaggac ggcggcgtgc tgcacgtgag cttcagctac 4501 cgctacgagg ccggccgcgt gatcggcgac ttcaaggtgg tgggcaccgg cttccccgag 4561 gacagcgtga tcttcaccga caagatcatc cgcagcaacg ccaccgtgga gcacctgcac 4621 cccatgggcg ataacgtgct ggtgggcagc ttcgcccgca ccttcagcct gcgcgacggc 4681 ggctactaca gcttcgtggt ggacagccac atgcacttca agagcgccat ccaccccagc 4741 atcctgcaga acgggggccc catgttcgcc ttccgccgcg tggaggagct gcacagcaac 4801 accgagctgg gcatcgtgga gtaccagcac gccttcaaga cccccatcgc cttcgccaga 4861 tcccgcgctc agtcgtccaa ttctgccgtg gacggcaccg ccggacccgg ctccaccgga 4921 tctcgcgagg gcagaggaag tcttctaaca tgcggtgacg tggaggagaa tcccggccct 4981 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta 5041 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac 5101 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac 5161 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag 5221 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt 5281 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag 5341 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc 5401 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg 5461 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc 5521 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga 5581 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct 5641 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt 5701 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg 5761 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact 5821 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct 5881 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg 5941 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc 6001 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc 6061 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctc 6121 cgccttcgcc ctcagacgag tcggatctcc ctttggccgc ctccccgcct ggtaccttta 6181 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga 6241 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct 6301 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 6361 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 6421 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt 6481 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt 6541 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 6601 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 6661 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt 6721 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct 6781 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg 6841 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 6901 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 6961 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 7021 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 7081 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 7141 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 7201 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 7261 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 7321 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 7381 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 7441 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 7501 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 7561 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 7621 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 7681 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 7741 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 7801 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 7861 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 7921 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 7981 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 8041 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 8101 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 8161 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 8221 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 8281 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 8341 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 8401 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 8461 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 8521 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 8581 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 8641 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 8701 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 8761 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 8821 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 8881 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 8941 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 9001 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 9061 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 9121 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 9181 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 9241 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga 9301 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat 9361 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat 9421 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca 9481 agctg //