LOCUS V045325 9785 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V045325 VERSION V045325 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..1920 /note="central polypurine tract and central termination sequence of HIV-1" /label="cPPT/CTS" promoter 2010..2213 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 2302..3852 /label="CHRND(NM_000751)" /note="CHRND(NM_000751)" /gene="CHRND" CDS 3889..3954 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 3955..4011 /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 porcine teschovirus-1 polyprotein" /transl_table=1 /translation="ATNFSLLKQAGDVEENPGP" /label="P2A" CDS 4012..4707 /codon_start=1 /note="mammalian codon-optimized" /product="red fluorescent protein from Entacmaea quadricolor" /transl_table=1 /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC DLPSKLGHR*" /label="TurboRFP" promoter 4755..4966 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4979..5247 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 5274..5873 /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 5880..6468 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6542..6775 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6847..6968 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6987..7122 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7160..7176) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7184..7200 /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(7208..7238) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7253..7274 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7562..8150) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8321..9181) /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(9182..9286) /gene="bla" /label="AmpR promoter" primer_bind 9760..9776 /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 attacaaaat tcaaaatttt 1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca 1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc 2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat 2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag 2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct 2281 agagctagcg aattcgccac catggagggg ccagtgctga cactggggct gctggctgcc 2341 ctggcggtgt gtggcagctg ggggctgaac gaggaggagc ggctgatccg gcacctgttt 2401 caagagaagg gctacaacaa ggagctccgg cccgtggcac acaaagagga gagtgtggac 2461 gttgccctgg ccctcacact ctccaacctc atctccctga aagaagttga ggagaccctc 2521 actaccaatg tgtggataga gcacggctgg acagacaacc ggctgaagtg gaatgctgaa 2581 gaatttggaa acatcagtgt cctgcgcctc cccccggaca tggtgtggct cccagagatt 2641 gtgctggaga acaacaatga cggctccttc cagatctcct actcctgcaa cgtgcttgtc 2701 taccactacg gcttcgtgta ctggctgcca cctgccatct tccgctcctc ctgccccatc 2761 tctgtcacct atttcccctt cgactggcag aactgctccc tcaagttcag ttccctcaag 2821 tatacggcca aagagatcac cctgagcctg aaacaggatg ccaaggagaa ccgcacctac 2881 cccgtggagt ggatcatcat tgatcctgaa ggcttcacag agaacgggga gtgggagata 2941 gtccaccggc cggccagggt caacgtggac cccagagccc ctctggacag ccccagccgc 3001 caggacatca ccttctacct catcatccgc cgcaagcccc tcttctacat catcaacatc 3061 ctggtgccct gcgtgctcat ctccttcatg gtcaacctgg tcttctacct accggctgac 3121 agtggtgaga agacatcagt ggccatctcg gtgctcctgg ctcagtctgt cttcctgctg 3181 ctcatctcca agcgtctgcc tgccacatcc atggccatcc cccttatcgg caagttcctg 3241 ctcttcggca tggtgctggt caccatggtt gtggtgatct gtgtcatcgt gctcaacatc 3301 cacttccgaa cacccagcac ccatgtgctg tctgaggggg tcaagaagct cttcctggag 3361 accctgccgg agctcctgca catgtcccgc ccagcagagg atggacccag ccctggggcc 3421 ctggtgcgga ggagcagctc cctgggatac atctccaagg ccgaggagta cttcctgctc 3481 aagtcccgca gtgacctcat gttcgagaag cagtcagagc ggcatgggct ggccaggcgc 3541 ctcaccactg cacgccggcc cccagcaagc tctgagcagg cccagcagga actcttcaat 3601 gagctgaagc cagctgtgga tggggcaaac ttcattgtta accacatgag ggaccagaac 3661 aattacaatg aggagaaaga cagctggaac cgagtggccc gcacagtgga ccgcctctgc 3721 ctgtttgtgg tgacgcctgt catggtggtg ggcacagcct ggatcttcct gcagggcgtt 3781 tacaaccagc caccacccca gccttttcct ggggacccct actcctacaa cgtgcaggac 3841 aagcgcttca tcggtggagg cggatcagga ggcggtggat ctctcgagga ctacaaagac 3901 catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga caaggctact 3961 aacttcagcc tgctgaagca ggctggagac gtggaggaga accctggacc tatgagcgag 4021 ctgatcaagg agaacatgca catgaagctg tacatggagg gcaccgtgaa caaccaccac 4081 ttcaagtgca catccgaggg cgaaggcaag ccctacgagg gcacccagac catgaagatc 4141 aaggtggtcg agggcggccc tctccccttc gccttcgaca tcctggctac cagcttcatg 4201 tacggcagca aagccttcat caaccacacc cagggcatcc ccgacttctt taagcagtcc 4261 ttccctgagg gcttcacatg ggagagaatc accacatacg aagacggggg cgtgctgacc 4321 gctacccagg acaccagctt ccagaacggc tgcatcatct acaacgtcaa gatcaacggg 4381 gtgaacttcc catccaacgg ccctgtgatg cagaagaaaa cacgcggctg ggaggccaac 4441 accgagatgc tgtaccccgc tgacggcggc ctgagaggcc acagccagat ggccctgaag 4501 ctcgtgggcg ggggctacct gcactgctcc ttcaagacca catacagatc caagaaaccc 4561 gctaagaacc tcaagatgcc cggcttccac ttcgtggacc acagactgga aagaatcaag 4621 gaggccgaca aagagaccta cgtcgagcag cacgagatgg ctgtggccaa gtactgcgac 4681 ctccctagca aactggggca cagatgagga tccgcggccg cgaaggatct gcgatcgctc 4741 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg 4801 ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt 4861 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt 4921 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg 4981 ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt 5041 gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt 5101 aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag 5161 actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc 5221 gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagatgaccg 5281 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc 5341 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca 5401 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca 5461 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg 5521 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc 5581 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt 5641 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg 5701 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct 5761 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg 5821 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgagtcgaca 5881 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc 5941 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta 6001 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt 6061 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg 6121 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta 6181 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt 6241 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg 6301 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca 6361 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc 6421 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggtaccttta 6481 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga 6541 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct 6601 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 6661 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 6721 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt 6781 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt 6841 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 6901 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 6961 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt 7021 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct 7081 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg 7141 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 7201 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 7261 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 7321 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 7381 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 7441 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 7501 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 7561 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 7621 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 7681 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 7741 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 7801 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 7861 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 7921 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 7981 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 8041 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 8101 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 8161 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 8221 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 8281 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 8341 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 8401 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 8461 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 8521 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 8581 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 8641 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 8701 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 8761 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 8821 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 8881 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 8941 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 9001 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 9061 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 9121 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 9181 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 9241 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 9301 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 9361 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca 9421 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc 9481 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag 9541 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga 9601 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat 9661 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat 9721 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca 9781 agctg //