LOCUS V040813 8526 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040813 VERSION V040813 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..2803 /label="GEMIN6(NM_024775)" /note="GEMIN6(NM_024775)" /gene="GEMIN6" CDS 2810..2875 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 2919..3130 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3143..3411 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 3446..4153 /codon_start=1 /note="mammalian codon-optimized" /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /transl_table=1 /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" /label="mCherry" CDS 4163..4216 /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 4217..4615 /codon_start=1 /gene="Aspergillus terreus BSD" /note="confers resistance to blasticidin" /product="blasticidin S deaminase" /transl_table=1 /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG*" /label="BSD" misc_feature 4622..5209 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5283..5516 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5588..5709 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5728..5863 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(5901..5917) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5925..5941 /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(5949..5979) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5994..6015 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6303..6891) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7062..7922) /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(7923..8027) /gene="bla" /label="AmpR promoter" primer_bind 8501..8517 /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 ccatgagtga atggatgaag aaaggcccct tagaatggca 2341 agattacatt tacaaagagg tccgagtgac agccagtgag aagaatgagt ataaaggatg 2401 ggttttaact acagacccag tctctgccaa tattgtcctt gtgaacttcc ttgaagatgg 2461 cagcatgtct gtgaccggaa ttatgggaca tgctgtgcag actgttgaaa ctatgaatga 2521 aggggaccat agagtgaggg agaagctgat gcatttgttc acgtctggag actgcaaagc 2581 atacagccca gaggatctgg aagagagaaa gaacagccta aagaaatggc ttgagaagaa 2641 ccacatcccc atcactgaac agggagacgc tccaaggact ctctgtgtgg ctggggtcct 2701 gactatagac ccaccatatg gtccagaaaa ttgcagcagc tctaatgaga ttattctgtc 2761 gcgtgttcag gatcttattg aaggacatct tacagcttcc caaggatccg actacaaaga 2821 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtagcg 2881 gccgcgaagg atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac 2941 agtccccgag aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg 3001 cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg 3061 agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc 3121 cagaacacag ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac 3181 ctgaggccgc catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc 3241 ctgaactgcg tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc 3301 ggcgctccct tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct 3361 tgctcaactc tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg 3421 accggcgcct acgctagacg ccaccatggt gagcaagggc gaggaggata acatggccat 3481 catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt 3541 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa 3601 ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta 3661 cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt 3721 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt 3781 gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac 3841 caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc 3901 cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct 3961 gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt 4021 gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga 4081 ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga 4141 cgagctgtac aagggatctc gcgagggcag aggaagtctt ctaacatgcg gtgacgtgga 4201 ggagaatccc ggccctatgg ccaagccttt gtctcaagaa gaatccaccc tcattgaaag 4261 agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg ccagcgcagc 4321 tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta ctgggggacc 4381 ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca acctgacttg 4441 tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac ggtgccgaca 4501 ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg atggacagcc 4561 gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg gctaagtcga 4621 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 4681 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 4741 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 4801 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 4861 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 4921 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 4981 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct 5041 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 5101 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 5161 ccgccttcgc cctcagacga gtcggatctc cctttggccg cctccccgcc tggtaccttt 5221 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg 5281 actggaaggg ctaattcact cccaacgaag ataagatctg ctttttgctt gtactgggtc 5341 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct 5401 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga 5461 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag 5521 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag 5581 tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa 5641 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa 5701 tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccag ttccgcccat 5761 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc 5821 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagactt ttgcagagac 5881 ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca 5941 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg 6001 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg 6061 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc 6121 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 6181 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 6241 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 6301 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 6361 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 6421 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 6481 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 6541 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 6601 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 6661 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 6721 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt 6781 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 6841 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 6901 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 6961 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 7021 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 7081 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 7141 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 7201 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 7261 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 7321 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 7381 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 7441 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 7501 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 7561 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 7621 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 7681 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 7741 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 7801 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 7861 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 7921 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 7981 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 8041 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg 8101 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac 8161 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc 8221 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca 8281 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg 8341 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga 8401 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga 8461 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc 8521 aagctg //