LOCUS V031957 9746 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V031957 VERSION V031957 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..3696 /label="G3bp1(NM_013716)" /note="G3bp1(NM_013716)" /gene="G3bp1" CDS 3706..3762 /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 3763..4473 /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" promoter 4515..4726 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4739..5007 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 5040..5834 /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 5841..6429 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6503..6736 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6808..6929 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6948..7083 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7121..7137) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7145..7161 /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(7169..7199) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7214..7235 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7523..8111) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8282..9142) /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(9143..9247) /gene="bla" /label="AmpR promoter" primer_bind 9721..9737 /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 catggttatg gagaaaccca gtcccctgct ggtcgggcgg 2341 gagtttgtgc gacagtacta cactctgctg aaccaggccc cggacatgtt gcacagattt 2401 tatggaaaga actcttccta tgcccacggg ggcttggatt ccaacgggaa gccagcagat 2461 gcagtctacg ggcagaagga aatccacagg aaagtgatgt cacaaaactt caccaactgc 2521 cacaccaaga tccgccacgt ggatgctcac gcaactctga atgacggggt ggtggtccaa 2581 gtgatggggc tactgtccaa caacaaccag gctctgcgga ggttcatgca gacctttgtc 2641 cttgctcctg agggctctgt tgccaacaaa ttctatgttc acaacgacat cttcaggtac 2701 caagatgagg tcttcggtgg ctttgtcaca gagcctcaag aggaatccga ggaagaagta 2761 gaagaacctg aagaaagaca gcagacacca gaggtggtgc ctgatgattc tggaactttc 2821 tatgatcaga ctgtcagcaa tgacttggag gagcatttag aggagcctgt agtggaacca 2881 gaaccggagc cagagccgga gccggagccg gaacctgtgt ccgacattca agaggacaag 2941 cctgaggctg cattggaaga agctgctccc gacgatgtgc agaagagcac ttccccggcc 3001 ccggctgatg tggccccggc acaggaggac ttgaggacgt tttcttgggc atctgtgacg 3061 agtaagaacc ttcctcccag tggggctgtt ccagtgacgg ggacaccacc tcatgtggtc 3121 aaagtgccag cgtcacagcc ccgtccagaa tctaaacctg actctcagat cccaccacaa 3181 aggcctcaga gagatcagag agttcgagag cagcgaatca atatccctcc tcagagagga 3241 ccccgaccaa tccgtgaggc tggtgaacca ggagatgtgg aacctcgaag gatggtgagg 3301 caccctgaca gtcaccagct cttcattggg aacctgccac atgaggttga caagtcggaa 3361 ctgaaggatt ttttccaaaa ttttggcaat gtggtggagc tgcgcatcaa cagtggtggg 3421 aagttaccca atttcggttt tgttgtgttt gatgattctg aacctgttca gaaggtcctt 3481 agtaataggc ccatcatgtt ccgaggcgcg gtccgcctga atgtggaaga gaagaagact 3541 cgagctgcca gagaaggcga ccgcagggat aaccgccttc ggggacctgg aggcccccgt 3601 ggtgggccga gtggtgggat gagaggccct ccccgcggag gcatggtgca gaaaccaggc 3661 tttggcgtgg gcagggggat tacaactcca aggcagggat ccggcgctac taacttcagc 3721 ctgctgaagc aggctggtga cgtggaggag aaccctggac ctatggtgag caagggcgag 3781 gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc 3841 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc 3901 cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg 3961 tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac 4021 tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac 4081 ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag 4141 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg 4201 ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc 4261 aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac 4321 aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc 4381 acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac 4441 tccaccggcg gcatggacga gctgtacaag taagcggccg cgaaggatct gcgatcgctc 4501 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg 4561 ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt 4621 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt 4681 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg 4741 ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt 4801 gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt 4861 aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag 4921 actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc 4981 gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc taggccacca 5041 tggctgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg 5101 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag 5161 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc 5221 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc 5281 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg 5341 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc 5401 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca 5461 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag 5521 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg 5581 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg 5641 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca 5701 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc 5761 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg 5821 acgagttctt ctgagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg 5881 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt 5941 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc 6001 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt 6061 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga 6121 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct 6181 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat 6241 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct 6301 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc 6361 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg 6421 cctccccgcc tggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac 6481 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaaa ataagatctg 6541 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc 6601 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg 6661 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg 6721 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca 6781 aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa 6841 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt 6901 ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa 6961 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 7021 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag 7081 gcctagactt ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt 7141 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag 7201 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt 7261 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag 7321 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 7381 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 7441 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 7501 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 7561 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 7621 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 7681 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 7741 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 7801 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 7861 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 7921 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct 7981 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 8041 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa 8101 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa 8161 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt 8221 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca 8281 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 8341 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc 8401 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa 8461 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc 8521 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca 8581 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat 8641 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag 8701 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac 8761 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt 8821 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt 8881 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc 8941 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat 9001 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca 9061 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga 9121 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg 9181 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg 9241 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga 9301 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg 9361 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg 9421 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct 9481 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa 9541 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc 9601 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag 9661 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt 9721 gtaaaacgac ggccagtgcc aagctg //