LOCUS V028588 8644 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V028588 VERSION V028588 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..2791 /label="Cep19(NM_025892)" /note="Cep19(NM_025892)" /gene="Cep19" promoter 2842..3053 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3066..3334 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 3369..4076 /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 4086..4139 /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 4140..4739 /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 4740..5327 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5401..5634 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5706..5827 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 5846..5981 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6019..6035) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6043..6059 /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(6067..6097) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6112..6133 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6421..7009) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7180..8040) /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(8041..8145) /gene="bla" /label="AmpR promoter" primer_bind 8619..8635 /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 ccatgaaata cattgccaag aaatgtggag ttaggttcca 2341 gcctccagct gtgatcttga tttatgagaa tgaaaccgaa ggaaagagcc gccagcgtat 2401 catgcctgtc cgaaactttt caaagttctc agattgcacc agagctgcgg aacagttaaa 2461 gaataaccca cggcacaaga gttacctgga acaggtgccc ctgaagcagc tggagaagct 2521 gtttgttttt ttgcgaggtt ccttgcaggg gcagagcttg gcagaaacaa tggaacagat 2581 tcggcgggaa acgaccatcg atcccgagga agacctgaac aaactggacg acaaggagct 2641 cgccaaaagg aagagcatca tggatgagct tttcgagaaa aatcagaaga gaaaggacga 2701 ccccactttt gtgtacgacg tcgaggtgga gttccctcag gatgaacagc tgctgtcctg 2761 cagctgggac acagcgtcag tggatgactt ctgaggatcc gcggccgcga aggatctgcg 2821 atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg 2881 ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta aactgggaaa 2941 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg 3001 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca cagctgaagc 3061 ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc cgccatccac 3121 gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg 3181 tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc ccttggagcc 3241 tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa ctctacgtct 3301 ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg cctacgctag 3361 acgccaccat ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc 3421 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg 3481 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc 3541 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg 3601 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt 3661 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc 3721 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg 3781 gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg 3841 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact 3901 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct 3961 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac 4021 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagggat 4081 ctcgcgaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat cccggcccta 4141 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac 4201 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc 4261 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca 4321 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga 4381 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt 4441 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc 4501 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca 4561 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg 4621 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg 4681 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgaa 4741 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc 4801 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta 4861 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt 4921 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg 4981 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta 5041 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt 5101 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg 5161 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca 5221 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtctcc 5281 gccttcgccc tcagacgagt cggatctccc tttggccgcc tccccgcctg gtacctttaa 5341 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac 5401 tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc 5461 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 5521 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 5581 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta 5641 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg 5701 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 5761 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 5821 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc 5881 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc 5941 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg 6001 cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 6061 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 6121 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 6181 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 6241 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 6301 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 6361 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 6421 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 6481 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 6541 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 6601 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 6661 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 6721 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 6781 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 6841 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac 6901 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 6961 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 7021 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 7081 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 7141 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 7201 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 7261 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 7321 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 7381 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 7441 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 7501 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 7561 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 7621 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 7681 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 7741 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 7801 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 7861 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 7921 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 7981 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 8041 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 8101 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 8161 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 8221 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 8281 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 8341 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 8401 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 8461 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 8521 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 8581 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa 8641 gctg //