LOCUS V042771 8427 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V042771 VERSION V042771 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 236..615 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 616..819 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 864..882 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 965..3106 /label="Cdh13(NM_019707)" /note="Cdh13(NM_019707)" /gene="Cdh13" CDS 3107..3172 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" CDS 3179..3232 /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 3257..3976 /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" polyA_signal 4028..4252 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 4298..4726 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 4740..5069 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 5136..5930 /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 6104..6225 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6274..6290) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6298..6314 /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(6322..6352) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6367..6388 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(6676..7261) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7432..8292) /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(8293..8397) /gene="bla" /label="AmpR promoter" ORIGIN 1 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat 61 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc 121 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg 181 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat 241 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat 301 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac 361 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc 421 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg 481 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat 541 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc 601 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt 661 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac 721 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc 781 ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc 841 actgcttact ggcttatcga aattaatacg actcactata gggagaccca agctggctag 901 cgtttaaact taagcttggt accgagctcg gatccactag tccagtgtgg tggaattcgc 961 caccatgcag ccgagaactc cgctcaccct gtgcgtcctg ctgtcccagg tgctcctggt 1021 cacgtctgca gatgatctgg agtgcacccc tggattccag cggaaagtgt tacacatcca 1081 ccagcctgcc gaattcatcg aggaccagcc tgtcctaaac ttgaccttca atgactgcaa 1141 gggcaacgag aagctgcact acgaggtctc aagcccacac ttcaaggtga acagcgacgg 1201 caccttagtg gctctcagga acatcactgc ggtgggcagg accctgtttg tccatgcgag 1261 gactcctcat gctgaagaca tggcagaact cgtgattgtc gggggcaaag acatccaggg 1321 ctccttgcag gatatcttta aatttgcaag gacatctcct gtcccaagac aaaagaggtc 1381 cattgtggtg tcccccatct tgatcccaga gaaccagaga cagcccttcc ccagagatgt 1441 cggcaaggta gtcgatagcg acagacccga agggtccaag ttccggctca ccgggaaggg 1501 agtggatcaa gaccctaaag gaactttcag aatcaatgag aacacaggca gtgtctccgt 1561 gacacggacc ctggacagag aaacgatcgc tacttatcaa ctgtatgtgg aaaccacgga 1621 tgccagtggc aaaactctgg aagggccagt gcctctggaa gtcattgtga ttgaccagaa 1681 cgacaacaga cccatcttcc gggaaggccc ttacatcggc catgtcatgg aagggtcacc 1741 cacagggacc acggtgatgc ggatgacagc gtttgatgca gatgacccgg ctactgacaa 1801 tgctctcctg aggtacaaca tccgtcagca gacgcctgac aagccatctc ctaacatgtt 1861 ctacatcgat cccgagaaag gagacattgt caccgtggtg tcacctgcgc tgctggaccg 1921 ggagactctg gaaaacccca agtatgaact gatcatcgaa gctcaagata tggcaggatt 1981 ggatgtcgga ctgacaggca cagccacagc caccatcgtg atcgatgaca aaaatgatca 2041 ctcaccaaaa ttcaccaaga aagagtttca agccacagtg gaggagggag ctgtgggggt 2101 cattgtcaac ttgacagtgg aagacaaaga cgaccctacc acaggagcct ggagggctgc 2161 atacaccatc atcaatggaa atcctgggca gagcttcgag atccacacca acccacagac 2221 caacgagggc atgctctctg tggtcaagcc cctggactat gagatctctg cctttcacac 2281 cctgctgatc aaagtggaga atgaggaccc actggtaccc gacgtctcct atggccccag 2341 ctccacggca actgtccaca tcacagtcct ggatgtcaac gagggaccag tcttctaccc 2401 ggaccccatg atggtgacta aacaggagaa catctctgtg ggcagtgtgc tgctgacagt 2461 gaatgccaca gacccagact ccctgcaaca tcaaactatc aggtactcta tctacaagga 2521 cccagcaggc tggctgagta tcaaccccat caatggaacg gtggacacca cagctgtgtt 2581 ggaccgggag tccccattcg tccacaacag tgtgtacacg gccctgttcc tggccatcga 2641 cagtggcaac cctcctgcaa ctggcaccgg gactctgctg ataaccctag aggacattaa 2701 tgacaacgct cctgtcattt acccaactgt ggctgaggtc tgcgatgatg ccagaaacct 2761 cagcgtggtc attctggggg cttcggacaa ggaccttcac cccaacacag accccttcaa 2821 gtttgagatt cataaacaga cagtccctga taaagtctgg aagatctcca aaatcaacaa 2881 cacccacgcc ctcgtgagcc ttcttcagaa tctgaacaag gcaaactaca acctgcccat 2941 catggtgaca gattcaggga agccacccat gacgaacatc acggacctca gggtgcaggt 3001 gtgctcttgc aagaactcca aagtggactg caacggggct ggggccctgc acctcagcct 3061 cagcttgctg ctgctcttct ctctcctcag cttattgtca ggtctggact acaaagacca 3121 tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca agccgcggga 3181 gggcagagga agtctgctaa catgcggtga cgtcgaggag aatcctggcc cagcccggga 3241 tccaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat 3301 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga 3361 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc 3421 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta 3481 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca 3541 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt 3601 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg 3661 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc 3721 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg 3781 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct 3841 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa 3901 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga 3961 cgagctgtac aagtaaagcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca 4021 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc 4081 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg 4141 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg 4201 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag 4261 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta 4321 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg 4381 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa 4441 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc 4501 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt 4561 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca 4621 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc 4681 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg 4741 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 4801 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa 4861 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca 4921 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt 4981 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag 5041 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg 5101 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg 5161 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa 5221 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg 5281 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt 5341 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa 5401 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc 5461 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg 5521 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg 5581 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg 5641 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg 5701 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact 5761 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg 5821 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc 5881 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct 5941 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac 6001 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat 6061 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc 6121 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc 6181 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc 6241 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg 6301 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg 6361 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc 6421 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt 6481 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct 6541 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga 6601 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc 6661 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg 6721 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg 6781 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt 6841 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt 6901 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg 6961 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact 7021 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt 7081 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct 7141 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac 7201 cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 7261 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 7321 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 7381 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 7441 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 7501 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 7561 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 7621 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 7681 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 7741 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 7801 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 7861 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 7921 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 7981 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 8041 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 8101 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 8161 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 8221 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 8281 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 8341 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 8401 atttccccga aaagtgccac ctgacgt //