LOCUS V044180 8947 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044180 VERSION V044180 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..3007 /label="TMED9(NM_017510)" /note="TMED9(NM_017510)" /gene="TMED9" CDS 3014..3040 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3044..3070 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3074..3100 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" promoter 3145..3356 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 3369..3637 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 3672..4379 /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 4389..4442 /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 4443..5042 /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 5043..5630 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5704..5937 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6009..6130 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6149..6284 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6322..6338) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6346..6362 /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(6370..6400) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6415..6436 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6724..7312) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7483..8343) /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(8344..8448) /gene="bla" /label="AmpR promoter" primer_bind 8922..8938 /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 ccatggctgt ggagctgggc gtgctgctcg tccggccccg 2341 gcccggaacc gggctgggta gagtgatgcg gaccctcctg ctggtgctgt ggctggcgac 2401 gcgcggaagc gcgctctact ttcacatcgg agagacggag aagaagtgct ttattgagga 2461 gatcccggac gagaccatgg tcataggaaa ctaccggacg cagctgtatg acaagcagcg 2521 ggaggagtac cagccggcca ccccggggct tggcatgttt gtggaggtga aggacccaga 2581 ggacaaggtc atcctggccc ggcagtatgg ctccgagggc aggttcactt tcacttccca 2641 tacccctggt gagcaccaga tctgtcttca ctccaattcc accaagttct ccctctttgc 2701 tggaggcatg ctgagagttc acctggacat ccaggtaggt gaacatgcca atgactatgc 2761 agaaattgct gctaaagaca agttgagtga gttgcagcta cgagtgcgac agctggtgga 2821 acaagtggag cagatccaga aagagcagaa ctaccagcgg tggcgagagg agcgcttccg 2881 gcagaccagt gagagcacca accagcgggt gctgtggtgg tccattctgc agaccctcat 2941 cctcgtggcc atcggtgtct ggcagatgcg gcacctcaag agcttctttg aagccaagaa 3001 gcttgtggga tcctacccat acgacgtacc tgattatgca acttatcctt atgatgtacc 3061 tgattatgct agctatcctt atgatgtccc agactacgca taggcggccg cgaaggatct 3121 gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt 3181 tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg 3241 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa 3301 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga 3361 agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc 3421 cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg 3481 ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga 3541 gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg 3601 tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc 3661 tagacgccac catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca 3721 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg 3781 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg 3841 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct 3901 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca 3961 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct 4021 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg 4081 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc 4141 ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc 4201 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg 4261 cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg 4321 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagg 4381 gatctcgcga gggcagagga agtcttctaa catgcggtga cgtggaggag aatcccggcc 4441 ctatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg 4501 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg 4561 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg 4621 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg 4681 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg 4741 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg 4801 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg 4861 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc 4921 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg 4981 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct 5041 gaaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg 5101 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc 5161 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt 5221 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca 5281 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc 5341 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc 5401 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc 5461 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc 5521 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc 5581 tccgccttcg ccctcagacg agtcggatct ccctttggcc gcctccccgc ctggtacctt 5641 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg 5701 gactggaagg gctaattcac tcccaacgaa aataagatct gctttttgct tgtactgggt 5761 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc 5821 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg 5881 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta 5941 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga 6001 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 6061 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 6121 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca 6181 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc 6241 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctagact tttgcagaga 6301 cggcccaaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 6361 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 6421 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 6481 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 6541 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 6601 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 6661 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 6721 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 6781 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 6841 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 6901 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 6961 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 7021 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 7081 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 7141 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 7201 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 7261 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 7321 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 7381 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 7441 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 7501 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 7561 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 7621 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 7681 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 7741 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 7801 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 7861 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 7921 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 7981 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 8041 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 8101 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 8161 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 8221 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 8281 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 8341 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 8401 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 8461 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 8521 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca 8581 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc 8641 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc 8701 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag 8761 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg 8821 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 8881 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc 8941 caagctg //