LOCUS V027594 8501 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V027594 VERSION V027594 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 50..429 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 430..633 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 775..2427 /label="GLDN(NM_181789)" /note="GLDN(NM_181789)" /gene="GLDN" CDS 2443..2499 /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 2500..3210 /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" misc_feature 3277..3865 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 3931..3979 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 4181..4609 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4623..4952 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 5019..5813 /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 5989..6110 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6159..6175) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6183..6199 /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(6207..6237) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6252..6273 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6561..7149) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7320..8180) /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(8181..8285) /gene="bla" /label="AmpR promoter" ORIGIN 1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 601 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg 661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg 721 actctagagc tagcgtttaa acttaagctt ggtaccgagc tcggatccgc caccatggcc 781 cgaggcgctg agggaggccg tggggacgcg ggttggggcc tgcgtggcgc cctggcggcc 841 gtggcgctgc tctcggcgct caacgctgcg ggcacggtgt tcgcgctgtg ccagtggcgc 901 gggctgagct cggcgctgcg ggctttggag gcgcagcggg gccgggagca gcgcgaggac 961 agtgccctgc gctccttcct ggccgagttg agccgcgcgc cgcgcggggc gtccgcacca 1021 ccccaagacc cggccagctc agctcgcaac aagcgcagcc acagcggcga gcccgcgccg 1081 catatccgcg ccgagagcca tgacatgctg atgatgatga cctactccat ggtgccgatc 1141 cgagtgatgg tggacctgtg caacagcacc aagggcatct gcctcacagg accttctgga 1201 ccaccaggac ctccgggagc cggcgggttg ccaggacaca acggattgga tggacagcct 1261 ggtcctcagg gcccaaaagg agaaaaagga gcaaatggaa aaagaggaaa aatggggata 1321 cctggagctg caggaaatcc aggggaaagg ggagaaaagg gagaccatgg tgaactgggc 1381 ctgcagggaa atgagggccc accagggcag aagggagaaa agggtgacaa aggagatgtg 1441 tccaacgacg tgctcctggc aggtgccaaa ggtgaccaag gcccacccgg tccacctggg 1501 cccccaggcc ctccaggtcc tccagggccc cctggaagca gaagagccaa aggccctcgg 1561 cagccaagca tgttcaacgg ccagtgccca ggtgagactt gtgccatacc aaatgatgat 1621 accttggttg gaaaagctga tgagaaagcc agtgaacacc attccccaca agcagaatcc 1681 atgatcactt ccattggaaa cccagtgcaa gtactgaaag tgacagagac atttgggact 1741 tggataagag agtctgctaa caagagtgat gaccggattt gggtgacaga gcatttttca 1801 ggcatcatgg ttaaggaatt caaggatcag ccctcacttc tgaatggcag ttacacgttc 1861 atccaccttc catactattt ccatggctgt gggcacgttg tttacaacaa ctctctctac 1921 taccacaaag ggggttctaa taccctagtg agatttgaat ttggccagga aacatcccaa 1981 actctgaagc ttgaaaatgc cttgtatttt gatcgaaaat acctttttgc aaattccaaa 2041 acttacttca atctagctgt agatgaaaag ggcctttgga ttatctatgc gtcaagtgtg 2101 gacggctcga gcattcttgt agcacaactg gatgagagga cattctcagt ggtgcaacac 2161 gtcaatacca cgtaccctaa atccaaggct ggcaacgcct tcattgcccg aggaatcctc 2221 tatgtcacag acaccaaaga tatgagggtc acatttgcct ttgatttgtt aggagggaaa 2281 cagatcaatg caaactttga tttaagaact tcccagtctg ttcttgccat gttagcatac 2341 aacatgagag atcagcattt atattcatgg gaagatggcc atttaatgct ttatcctgtg 2401 cagtttttgt caactacctt aaatcaggaa ttcggaagcg gagctactaa cttcagcctg 2461 ctgaagcagg ctggcgacgt ggaggagaac cctggaccta tggtgagcaa gggcgaggag 2521 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg 2581 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag 2641 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc 2701 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac 2761 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc 2821 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg 2881 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc 2941 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag 3001 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag 3061 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc 3121 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc 3181 accggcggca tggacgagct gtacaagtaa ctcgagtcta gagggcccct taagggttcg 3241 atccctaccg gttagtaatg agtttgatat ctcgacaatc aacctctgga ttacaaaatt 3301 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct 3361 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg 3421 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc 3481 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt 3541 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc 3601 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg 3661 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg 3721 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc 3781 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg 3841 atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa acacggaagg 3901 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg 3961 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat 4021 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac 4081 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc 4141 atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca 4201 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 4261 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 4321 caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg gcacctcgac 4381 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt 4441 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 4501 acaacactca accctatctc ggtctattct tttgatttat aagggatttt ggggatttcg 4561 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga 4621 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 4681 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca 4741 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc 4801 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt 4861 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag 4921 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt 4981 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc 5041 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga 5101 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt 5161 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat 5221 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg 5281 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg 5341 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc 5401 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga 5461 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag 5521 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc 5581 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg 5641 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata 5701 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg 5761 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac 5821 tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat 5881 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg 5941 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt 6001 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt 6061 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt 6121 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga 6181 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc 6241 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc 6301 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 6361 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 6421 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 6481 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 6541 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 6601 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 6661 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 6721 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt 6781 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 6841 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 6901 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 6961 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc 7021 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 7081 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 7141 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 7201 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 7261 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 7321 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 7381 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 7441 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 7501 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 7561 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 7621 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 7681 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 7741 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 7801 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 7861 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 7921 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 7981 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 8041 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 8101 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 8161 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 8221 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 8281 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc 8341 ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc 8401 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa 8461 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g //