LOCUS V019724 9313 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019724 VERSION V019724 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 2294..3484 /label="DNAJA1(NM_001539)" /note="DNAJA1(NM_001539)" /gene="DNAJA1" promoter 3534..3745 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3758..4026 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4061..4753 /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" CDS 4754..4807 /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 4808..5407 /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 5408..5996 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6070..6303 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6375..6496 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6515..6650 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6688..6704) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6712..6728 /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(6736..6766) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6781..6802 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7090..7678) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7849..8709) /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(8710..8814) /gene="bla" /label="AmpR promoter" primer_bind 9288..9304 /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 agagctagcc accatggtga aagaaacaac ttactacgat gttttggggg tcaaacccaa 2341 tgctactcag gaagaattga aaaaggctta taggaaactg gctttgaagt accatcctga 2401 taagaaccca aatgaaggag agaagtttaa acagatttct caagcttacg aagttctctc 2461 tgatgcaaag aaaagggaat tatatgacaa aggaggagaa caggcaatta aagagggtgg 2521 agcaggtggc ggttttggct cccccatgga catctttgat atgttttttg gaggaggagg 2581 aaggatgcag agagaaagga gaggtaaaaa tgttgtacat cagctctcag taaccctaga 2641 agacttatat aatggtgcaa caagaaaact ggctctgcaa aagaatgtga tttgtgacaa 2701 atgtgaaggt agaggaggta agaaaggagc agtagagtgc tgtcccaatt gccgaggtac 2761 tggaatgcaa ataagaattc atcagatagg acctggaatg gttcagcaaa ttcagtctgt 2821 gtgcatggag tgccagggcc atggggagcg gatcagtcct aaagatagat gtaaaagctg 2881 caacggaagg aagatagttc gagagaagaa aattttagaa gttcatattg acaaaggcat 2941 gaaagatggc cagaagataa cattccatgg tgaaggagac caagaaccag gactggagcc 3001 aggcgatatt atcattgtgt tagatcagaa ggaccatgct gtttttactc gacgaggaga 3061 agaccttttc atgtgtatgg acatacagct cgttgaagca ctgtgtggct tccagaagcc 3121 aatatctact cttgacaacc gaaccatcgt catcacctct catccaggtc agattgtcaa 3181 gcatggagat atcaagtgtg tactaaatga aggcatgcca atttatcgta gaccatatga 3241 aaagggtcgc ctaatcatcg aatttaaggt aaactttcct gagaatggct ttctctctcc 3301 tgataaactg tctttgctgg aaaaactcct acccgagagg aaggaagtgg aagagactga 3361 tgagatggac caagtagaac tggtggactt tgatccaaat caggaaagac ggcgccacta 3421 caatggagaa gcatatgagg atgatgaaca tcatcccaga ggtggtgttc agtgtcagac 3481 ctcttaagga tccgcggccg caaggatctg cgatcgctcc ggtgcccgtc agtgggcaga 3541 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc 3601 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt 3661 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg 3721 caacgggttt gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg 3781 cgcccgccgc cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc 3841 cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag 3901 accgggcctt tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct 3961 ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta 4021 cagatccaag ctgtgaccgg cgcctacgct agacgccacc atgagcgagc tgatcaagga 4081 gaacatgcac atgaagctgt acatggaggg caccgtgaac aaccaccact tcaagtgcac 4141 atccgagggc gaaggcaagc cctacgaggg cacccagacc atgaagatca aggtggtcga 4201 gggcggccct ctccccttcg ccttcgacat cctggctacc agcttcatgt acggcagcaa 4261 agccttcatc aaccacaccc agggcatccc cgacttcttt aagcagtcct tccctgaggg 4321 cttcacatgg gagagaatca ccacatacga agacgggggc gtgctgaccg ctacccagga 4381 caccagcttc cagaacggct gcatcatcta caacgtcaag atcaacgggg tgaacttccc 4441 atccaacggc cctgtgatgc agaagaaaac acgcggctgg gaggccaaca ccgagatgct 4501 gtaccccgct gacggcggcc tgagaggcca cagccagatg gccctgaagc tcgtgggcgg 4561 gggctacctg cactgctcct tcaagaccac atacagatcc aagaaacccg ctaagaacct 4621 caagatgccc ggcttccact tcgtggacca cagactggaa agaatcaagg aggccgacaa 4681 agagacctac gtcgagcagc acgagatggc tgtggccaag tactgcgacc tccctagcaa 4741 actggggcac agagagggca gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc 4801 cggccctatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag 4861 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga 4921 tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg 4981 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac 5041 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt 5101 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc 5161 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg 5221 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc 5281 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt 5341 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg 5401 tgcctgaaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta 5461 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc 5521 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga 5581 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac 5641 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc 5701 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc 5761 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg 5821 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc 5881 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc 5941 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg 6001 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa 6061 aggggggact ggaagggcta attcactccc aacgaaaata agatctgctt tttgcttgta 6121 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc 6181 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt 6241 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta 6301 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat 6361 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca 6421 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac 6481 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccagttc 6541 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc 6601 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg 6661 cagagacggc ccaaattcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc 6721 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 6781 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 6841 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 6901 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 6961 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 7021 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 7081 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 7141 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 7201 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 7261 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 7321 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 7381 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 7441 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 7501 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 7561 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 7621 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 7681 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 7741 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 7801 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 7861 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 7921 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 7981 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 8041 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 8101 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 8161 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 8221 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 8281 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 8341 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 8401 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 8461 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 8521 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 8581 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 8641 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 8701 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 8761 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 8821 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 8881 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct 8941 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 9001 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc 9061 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg 9121 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga 9181 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc 9241 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc 9301 cagtgccaag ctg //