LOCUS V046621 9902 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V046621 VERSION V046621 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 2317..3903 /label="Runx2(NM_001278483)" /note="Runx2(NM_001278483)" /gene="Runx2" CDS 3910..3936 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3940..3966 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" CDS 3970..3996 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /transl_table=1 /translation="YPYDVPDYA" /label="HA" promoter 4061..4272 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4285..4553 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" regulatory 4582..4591 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" /label="Kozak sequence" CDS 4612..5274 /codon_start=1 /product="green fluorescent protein 2 from Pontellina plumata, also known as ppluGFP2 (Shagin et al., 2004)" /transl_table=1 /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA" /label="CopGFP" CDS 5344..5397 /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 5398..5997 /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 5998..6585 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 6659..6892 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 6964..7085 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 7104..7239 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(7277..7293) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 7301..7317 /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(7325..7355) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 7370..7391 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7679..8267) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(8438..9298) /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(9299..9403) /gene="bla" /label="AmpR promoter" primer_bind 9877..9893 /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 ctgatacgaa ctcggaattc gccaccatgg cgtcaaacag cctcttcagt 2341 gcagtgacac cgtgtcagca aaacttcttt tgggatccga gcaccagccg gcgcttcagc 2401 cccccctcca gcagcctgca gcccggcaag atgagcgacg tgagcccggt agtggctgct 2461 cagcagcagc agcagcagca gcagcagcag cagcagcagc aacagcagca acagcagcag 2521 cagcagcagc aacagcagca gcagcagcag caggaggcgg ccgcagcagc agcagcggca 2581 gcggcggcgg cagcagcggc ggcagccgtg ccccggttga ggccgccgca cgacaaccgc 2641 accatggtgg agatcatcgc ggaccacccg gccgaactgg tccgcaccga cagccccaac 2701 ttcctgtgct ccgtgctgcc ctcgcactgg cggtgcaaca agaccctgcc cgtggccttc 2761 aaggttgtag ccctcggaga ggtacccgat gggaccgtgg ttaccgtcat ggccgggaat 2821 gatgagaact actctgccga gctacgaaat gcctctgctg ttatgaaaaa ccaagtggcc 2881 aggttcaacg atctgagatt tgtaggccgg agcggacgag gcaagagttt cactttgacc 2941 ataacggtct tcacaaatcc tccccaagtg gccacttacc acagagctat taaagtgaca 3001 gtggacggtc cccgggaacc aagaaggcac agacagaagc ttgatgactc taaacctagt 3061 ttgttctctg accgcctcag tgatttaggg cgcattcctc atcccagtat gagagtaggt 3121 gtcccgcctc agaacccacg gccctccctg aactcagcac caagtccttt taatccacaa 3181 ggacagagtc agattacaga tcccaggcag acacaatcct ccccaccgtg gtcctatgac 3241 cagtcttacc cctcttatct gagccagatg acgtccccat ccatccattc caccacgccg 3301 ctgtcttcca cacggggcac tgggctgcca gccatcaccg acgtacccag gcgtatttca 3361 gatgatgaca ctgccacctc tgacttctgc ctctggcctt cctctctcag taagaagagc 3421 caggcaggtg cttcagaact gggccctttt tcagacccca ggcagttccc aggcatttca 3481 tccctcactg agagccgctt ctccaaccca cgaatgcact atccagccac cttcacttac 3541 accccgccag tcacgtcagg catgtccctg ggcatgtccg ccaccactca ctaccacacg 3601 tacctgccac caccctaccc cggctcttcc caaagccaga gcggaccctt ccagaccagc 3661 agcactccat atctctacta tggtacttcg tcagcgtcct atcagttccc aatggtcccc 3721 gggggagacc ggtctccttc caggatggtc ccaccgtgca cgaccacctc gaatggcagc 3781 acgctattaa atccaaattt gcctaaccag aatgatggtg ttgacgctga tggaagccac 3841 agcagctccc caaccgtttt gaattctagc ggcagaatgg atgagtctgt ctggcgaccc 3901 tatctcgagt acccatacga cgtacctgat tatgcaactt atccttatga tgtacctgat 3961 tatgctagct atccttatga tgtcccagac tacgcatagg gatccctgcg atatccagcg 4021 cggccgcgaa ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc 4081 acagtccccg agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg 4141 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg 4201 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc 4261 gccagaacac agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct 4321 acctgaggcc gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc 4381 tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt 4441 ccggcgctcc cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg 4501 cttgctcaac tctacgtctt tgtttcgttt tctgttctgc gccgttacag atccaagctg 4561 tgaccggcgc ctacgctaga cgccaccatg gagagcgacg agagcggcct gcccgccatg 4621 gagatcgagt gccgcatcac cggcaccctg aacggcgtgg agttcgagct ggtgggcggc 4681 ggagagggca cccccaagca gggccgcatg accaacaaga tgaagagcac caaaggcgcc 4741 ctgaccttca gcccctacct gctgagccac gtgatgggct acggcttcta ccacttcggc 4801 acctacccca gcggctacga gaaccccttc ctgcacgcca tcaacaacgg cggctacacc 4861 aacacccgca tcgagaagta cgaggacggc ggcgtgctgc acgtgagctt cagctaccgc 4921 tacgaggccg gccgcgtgat cggcgacttc aaggtggtgg gcaccggctt ccccgaggac 4981 agcgtgatct tcaccgacaa gatcatccgc agcaacgcca ccgtggagca cctgcacccc 5041 atgggcgata acgtgctggt gggcagcttc gcccgcacct tcagcctgcg cgacggcggc 5101 tactacagct tcgtggtgga cagccacatg cacttcaaga gcgccatcca ccccagcatc 5161 ctgcagaacg ggggccccat gttcgccttc cgccgcgtgg aggagctgca cagcaacacc 5221 gagctgggca tcgtggagta ccagcacgcc ttcaagaccc ccatcgcctt cgccagatcc 5281 cgcgctcagt cgtccaattc tgccgtggac ggcaccgccg gacccggctc caccggatct 5341 cgcgagggca gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc cggccctatg 5401 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc 5461 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc 5521 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc 5581 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc 5641 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc 5701 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc 5761 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc 5821 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag 5881 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac 5941 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaaat 6001 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct 6061 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg 6121 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg 6181 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt 6241 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt 6301 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg 6361 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc 6421 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat 6481 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtctccgc 6541 cttcgccctc agacgagtcg gatctccctt tggccgcctc cccgcctggt acctttaaga 6601 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg 6661 gaagggctaa ttcactccca acgaaaataa gatctgcttt ttgcttgtac tgggtctctc 6721 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag 6781 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct 6841 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt 6901 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag 6961 aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 7021 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 7081 tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccagttcc gcccattctc 7141 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg 7201 agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc agagacggcc 7261 caaattcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc 7321 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct 7381 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 7441 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 7501 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 7561 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 7621 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 7681 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 7741 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 7801 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 7861 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 7921 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 7981 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 8041 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 8101 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct 8161 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 8221 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 8281 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 8341 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 8401 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 8461 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 8521 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 8581 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 8641 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 8701 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 8761 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 8821 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 8881 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 8941 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 9001 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 9061 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 9121 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 9181 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 9241 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 9301 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 9361 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 9421 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta 9481 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc 9541 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc 9601 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc 9661 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa 9721 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg 9781 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa 9841 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc 9901 tg //