LOCUS V044629 8890 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V044629 VERSION V044629 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..3018 /codon_start=1 /note="mammalian codon-optimized" /product="the original enhanced GFP (Yang et al., 1996)" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV LLEFVTAAGITLGMDELYK" /label="EGFP" CDS 3055..3816 /label="MEOX1(NM_004527)" /note="MEOX1(NM_004527)" /gene="MEOX1" promoter 3860..4071 /note="core promoter for human elongation factor EF-1α" /label="EF-1α core promoter" LTR 4084..4352 /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" /label="5' LTR (truncated)" CDS 4379..4978 /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 4985..5573 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" LTR 5647..5880 /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" /label="3' LTR (ΔU3)" polyA_signal 5952..6073 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin 6092..6227 /note="SV40 origin of replication" /label="SV40 ori" primer_bind complement(6265..6281) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6289..6305 /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(6313..6343) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6358..6379 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6667..7255) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7426..8286) /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(8287..8391) /gene="bla" /label="AmpR promoter" primer_bind 8865..8881 /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 catggtgagc aagggcgagg agctgttcac cggggtggtg 2341 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag 2401 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag 2461 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc 2521 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac 2581 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg 2641 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag 2701 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc 2761 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag 2821 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc 2881 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac 2941 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc 3001 atggacgagc tgtacaaggg cggcggaggg agtggaggag ggggaagtgg atccatggat 3061 cccgcggcca gcagctgcat gaggagcctc cagcccccag cccctgtctg gggctgcctt 3121 cgaaaccccc actcggaagg caatggggcc tcagggctac cccactaccc gcccaccccg 3181 ttctccttcc accagaaacc agacttcctg gcgacagcga cggcagcgta ccctgacttc 3241 tcagcctcct gcctggcagc caccccacac agcctgcccc aggaggagca catcttcact 3301 gagcagcacc ccgctttccc acagtccccc aactggcact tccctgtctc agacgcccgg 3361 cgcaggccca actcaggccc ggcagggggt tccaaggaaa tggggaccag cagcctgggc 3421 ctggtggaca ccacaggagg cccaggcgat gactacgggg tgcttgggag cactgccaat 3481 gagacagaga agaaatcatc caggcggaga aaggagagtt cagacaacca ggagaacaga 3541 gggaagccgg agggcagcag caaagcccgc aaggagagga cggccttcac caaggagcag 3601 ctgcgagagc tggaggcaga gtttgcccat cataactacc tgactcggct ccgcagatat 3661 gagattgcgg taaacctgga cctctctgag cgccaggtca aagtgtggtt ccagaaccga 3721 aggatgaagt ggaagcgtgt gaagggaggt cagcccatct cccccaatgg gcaggaccct 3781 gaggatgggg actccacagc ctctccaagt tcagagtagc ggccgcgaag gatctgcgat 3841 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 3901 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 3961 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 4021 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt 4081 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc 4141 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc 4201 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta 4261 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt 4321 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagat 4381 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg 4441 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg acccggaccg 4501 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat 4561 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag 4621 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc 4681 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc 4741 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag 4801 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga 4861 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga 4921 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgagt 4981 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 5041 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 5101 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 5161 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 5221 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 5281 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 5341 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct 5401 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 5461 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 5521 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggtac 5581 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg 5641 ggggactgga agggctaatt cactcccaac gaaaataaga tctgcttttt gcttgtactg 5701 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac 5761 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt 5821 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca 5881 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag 5941 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 6001 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 6061 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccagttccgc 6121 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc 6181 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag 6241 agacggccca aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct 6301 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 6361 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 6421 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 6481 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 6541 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 6601 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 6661 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 6721 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 6781 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 6841 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 6901 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 6961 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 7021 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 7081 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc 7141 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 7201 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 7261 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 7321 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 7381 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 7441 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 7501 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 7561 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 7621 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 7681 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 7741 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 7801 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 7861 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 7921 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 7981 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 8041 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 8101 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 8161 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 8221 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 8281 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 8341 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 8401 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa 8461 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg 8521 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca 8581 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc 8641 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt 8701 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg 8761 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag 8821 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag 8881 tgccaagctg //