LOCUS V041248 9803 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V041248 VERSION V041248 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..3679 /label="RORB(NM_006914)" /note="RORB(NM_006914)" /gene="RORB" CDS 3686..3751 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" promoter 3795..4006 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4019..4287 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4322..5029 /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 5039..5092 /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 5099..5893 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" misc_feature 5900..6488 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6560..6793 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6865..6986 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7005..7140 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7178..7194) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7202..7218 /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(7226..7256) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7271..7292 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7580..8168) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8339..9199) /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(9200..9304) /gene="bla" /label="AmpR promoter" primer_bind 9778..9794 /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 ccatgcgagc acaaattgaa gtgataccat gcaaaatttg 2341 tggcgataag tcctctggga tccactacgg agtcatcaca tgtgaaggct gcaagggatt 2401 ctttaggagg agccagcaga acaatgcttc ttattcctgc ccaaggcaga gaaactgttt 2461 aattgacaga acgaacagaa accgttgcca acactgccga ctgcagaagt gtcttgccct 2521 aggaatgtca agagatgctg tgaagtttgg gaggatgtcc aagaagcaaa gggacagcct 2581 gtatgctgag gtgcagaagc accagcagcg gctgcaggaa cagcggcagc agcagagtgg 2641 ggaggcagaa gcccttgcca gggtgtacag cagcagcatt agcaacggcc tgagcaacct 2701 gaacaacgag accagcggca cttatgccaa cgggcacgtc attgacctgc ccaagtctga 2761 gggttattac aacgtcgatt ccggtcagcc gtcccctgat cagtcaggac ttgacatgac 2821 tggaatcaaa cagataaagc aagaacctat ctatgacctc acatccgtac ccaacttgtt 2881 tacctatagc tctttcaaca atgggcagtt agcaccaggg ataaccatga ctgaaatcga 2941 ccgaattgca cagaacatca ttaagtccca tttggagaca tgtcaataca ccatggaaga 3001 gctgcaccag ctggcgtggc agacccacac ctatgaagaa attaaagcat atcaaagcaa 3061 gtccagggaa gcactgtggc aacaatgtgc catccagatc actcacgcca tccaatacgt 3121 ggtggagttt gcaaagcgga taacaggctt catggagctc tgtcaaaatg atcaaattct 3181 acttctgaag tcaggttgct tggaagtggt tttagtgaga atgtgccgtg ccttcaaccc 3241 attaaacaac actgttctgt ttgaaggaaa atatggagga atgcaaatgt tcaaagcctt 3301 aggttctgat gacctagtga atgaagcatt tgactttgca aagaatttgt gttccttgca 3361 gctgaccgag gaggagatcg ctttgttctc atctgctgtt ctgatatctc cagaccgagc 3421 ctggcttata gaaccaagga aagtccagaa gcttcaggaa aaaatttatt ttgcacttca 3481 acatgtgatt cagaagaatc acctggatga tgagaccttg gcaaagttaa tagccaagat 3541 accaaccatc acggcagttt gcaacttgca cggggagaag ctgcaggtat ttaagcaatc 3601 tcatccagag atagtgaata cactgtttcc tccgttatac aaggagctct ttaatcctga 3661 ctgtgccacc ggctgcaaag gatccgacta caaagaccat gacggtgatt ataaagatca 3721 tgacatcgac tacaaggatg acgatgacaa gtagcggccg cgaaggatct gcgatcgctc 3781 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg 3841 ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt 3901 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt 3961 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg 4021 ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt 4081 gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt 4141 aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag 4201 actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc 4261 gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagacgccac 4321 catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa 4381 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg 4441 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc 4501 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca 4561 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg 4621 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga 4681 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt 4741 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg 4801 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc 4861 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt 4921 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga 4981 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagg gatctcgcga 5041 gggcagagga agtcttctaa catgcggtga cgtggaggag aatcccggcc ctgccaccat 5101 ggctgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg 5161 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc 5221 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca 5281 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct 5341 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga 5401 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg 5461 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat 5521 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga 5581 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg 5641 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg 5701 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat 5761 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct 5821 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga 5881 cgagttcttc tgagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg 5941 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta 6001 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct 6061 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt 6121 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac 6181 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg 6241 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc 6301 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg 6361 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct 6421 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc 6481 ctccccgcgg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt 6541 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagata agatctgctt 6601 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa 6661 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt 6721 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg 6781 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag 6841 aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa 6901 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt 6961 ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc 7021 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg 7081 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc 7141 tagacttttg cagagacggc ccaaattcgt aatcatggtc atagctgttt cctgtgtgaa 7201 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct 7261 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc 7321 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg 7381 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc 7441 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag 7501 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa 7561 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc 7621 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc 7681 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg 7741 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt 7801 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc 7861 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc 7921 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag 7981 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg 8041 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa 8101 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 8161 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 8221 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 8281 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 8341 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 8401 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 8461 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 8521 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 8581 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 8641 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 8701 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 8761 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 8821 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 8881 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 8941 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 9001 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 9061 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 9121 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 9181 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 9241 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 9301 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat 9361 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg 9421 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg 9481 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc 9541 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac 9601 cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca 9661 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg 9721 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta 9781 aaacgacggc cagtgccaag ctg //