LOCUS V040475 6901 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040475 VERSION V040475 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 235..614 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 615..818 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 863..881 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 964..2391 /label="Gdf10(NM_145741)" /note="Gdf10(NM_145741)" /gene="Gdf10" CDS 2398..2463 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2501..2725 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2771..3199 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3213..3542 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3609..4403 /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 4577..4698 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4747..4763) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4771..4787 /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(4795..4825) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4840..4861 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5149..5734) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5905..6765) /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(6766..6870) /gene="bla" /label="AmpR promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc 961 accatggctc caggtcctgc tcggatcagc ttggggtccc agctgctgcc catggtgccg 1021 ctgctcctgc tgctgcgggg cgcaggctgc ggccacaggg gcccctcatg gtcctcattg 1081 ccctcggcag ctgccggtct gcagggggac agggactccc agcagtcacc cggggacgca 1141 gcagccgctc tgggcccagg cgcccaggac atggtcgcta tccacatgct caggctctat 1201 gagaagtaca accgaagagg tgctccaccg ggaggaggca acaccgtccg aagcttccgt 1261 gcccggctgg aaatgatcga ccaaaagcct gtgtatttct tcaacttgac ttccatgcaa 1321 gactcagaaa tgatcctcac agccgccttc cacttctact cagaacctcc acggtggccc 1381 cgggctcgtg aggtattctg caagccccga gctaagaacg catcctgccg cctcctgacc 1441 ccagggctgc ctgcacgctt gcacctaatc ttccgcagtc tttcccagaa caccgccact 1501 caggggctgc tccgcggggc catggccctg acgcctccac cacgtggcct gtggcaggcc 1561 aaggacatct cctcaatcat caaggctgcc cgaagggatg gagagctgct tctctctgct 1621 cagctggata ctggggagaa ggaccccgga gtgccacggc ccagttccca catgccctat 1681 atccttgtct acgccaatga cctggccatc tccgaaccca acagtgtagc agtgtcgcta 1741 cagagatacg acccatttcc agctggagac tttgagcctg gagcagcccc caacagctca 1801 gctgatcccc gcgtgcgcag ggcggctcag gtgtcaaaac ccctgcaaga caatgaactg 1861 ccggggctgg atgaaagacc agcgcctgcc ctgcatgccc agaatttcca caagcacgag 1921 ttctggtcca gtcctttccg ggcactgaaa ccccgcacgg ggcgcaaaga ccgcaagaag 1981 aaggaccagg acacattcac cgccgcctcc tctcaggtgc tggactttga cgagaagacg 2041 atgcagaaag ccaggaggcg gcagtgggat gagccccggg tctgctccag gaggtacctg 2101 aaggtggatt ttgcagacat cgggtggaat gaatggatca tctctcccaa atcctttgac 2161 gcctactact gtgctggggc ctgcgagttc cccatgccca agattgtccg cccatccaac 2221 catgccacca tccagagcat cgtcagagct gtgggcattg tccctggcat cccagagcca 2281 tgctgtgttc cagacaagat gaactccctt ggagtccttt tcctggatga aaatcggaat 2341 gcggttctga aggtgtaccc caatatgtcc gtagagacct gtgcctgtcg gctcgaggac 2401 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac 2461 aagtagtgag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag 2521 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact 2581 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt 2641 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat 2701 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg 2761 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc 2821 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc 2881 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg 2941 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca 3001 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc 3061 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct 3121 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa 3181 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc 3241 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt 3301 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 3361 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg 3421 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct 3481 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca 3541 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat 3601 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga 3661 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc 3721 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga 3781 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg 3841 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc 3901 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg 3961 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga 4021 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc 4081 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca 4141 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg 4201 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct 4261 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg 4321 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc 4381 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac 4441 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt 4501 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga 4561 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag 4621 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 4681 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta 4741 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat 4801 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt 4861 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta 4921 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc 4981 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 5041 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 5101 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 5161 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 5221 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 5281 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 5341 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 5401 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 5461 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 5521 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 5581 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 5641 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt ttgtttgcaa 5701 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 5761 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 5821 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 5881 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 5941 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 6001 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 6061 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 6121 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 6181 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 6241 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 6301 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 6361 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 6421 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 6481 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 6541 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 6601 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 6661 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 6721 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 6781 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 6841 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 6901 c //