LOCUS Exported 6959 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS F-Cdk8 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6959) AUTHORS Conaway RC, Sato S, Tomomori-Sato C, Yao T, Conaway JW TITLE The mammalian Mediator complex and its role in transcriptional regulation. JOURNAL Trends Biochem Sci. 2005 May . 30(5):250-5. PUBMED 15896743 REFERENCE 2 (bases 1 to 6959) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6959 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 907..930 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" primer_bind complement(2381..2398) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2387..2611 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2657..3085 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2744..2763) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2954..2975 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3094..3114) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3099..3428 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3279..3414 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3341..3360 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3477..4502 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" polyA_signal 4632..4753 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4669..4688) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4723..4742 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4802..4818) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4802..4818) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4815..4837) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4826..4842 /label=lac operator /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-beta-D-thiogalactopyranoside (IPTG)." promoter complement(4850..4880) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4895..4916 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5033..5050) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5204..5792) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5284..5303) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5963..6823) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 6586..6605 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6824..6928) /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 atgccagact acaaggacga tgatgacaag ctcgagatgg actatgactt taaagtgaag 961 ctgagcagcg agcgggagcg ggtcgaggac ctgtttgaat acgagggctg caaagttggc 1021 cgaggcactt atggtcacgt ctacaaagcc aagaggaaag atgggaagga tgataaagac 1081 tatgctttaa aacaaataga aggaactggg atctctatgt cggcatgtag agaaatagca 1141 ttacttcgag agcttaagca tccaaacgtc atttctcttc aaaaggtgtt tctgtctcat 1201 gctgatagga aggtgtggct tctgtttgac tatgctgaac atgacctctg gcatataatc 1261 aagtttcaca gagcttctaa agcaaacaag aagccagttc agttacctcg gggaatggtg 1321 aagtcactat tatatcagat cctagatggt attcactacc tgcatgctaa ctgggtgttg 1381 cacagagatt tgaaacctgc taatatttta gttatgggtg aaggtcctga gcgaggaaga 1441 gtaaaaattg ctgacatggg ctttgcccga ttatttaatt cacctttgaa gcctttagca 1501 gatttggatc cagtggttgt tacattctgg taccgagccc ctgaactact tcttggagca 1561 aggcattata ccaaagctat tgatatttgg gctatagggt gtatatttgc agaactacta 1621 acgtcagaac caatatttca ctgtcgacaa gaggacatca aaactagtaa tccttatcac 1681 catgaccagc tggacagaat attcaatgta atgggatttc ctgcagataa agattgggaa 1741 gatataaaaa agatgcctga acattcaaca ttaatgaaag atttcagaag aaatacgtat 1801 accaactgca gccttatcaa gtatatggaa aaacataaag ttaaaccaga tagtaaagca 1861 ttccacttgc ttcagaagct gcttaccatg gacccaataa agcgaattac ctcagaacag 1921 gctatgcagg acccctattt cttagaagac ccacttccta catcagacgt ttttgccggt 1981 tgtcaaatcc cttacccaaa acgagaattt ttaacggaag aagaacctga tgacaaagga 2041 gacaaaaaga accagcagca gcagcagggc aataaccaca ctaatggaac tggccaccca 2101 gggaatcaag acagcagtca cacacaggga cccccgttga agaaagtgag agttgttcct 2161 cctaccacta cctcaggtgg acttatcatg acctcagact atcagcgttc caatccacat 2221 gctgcctatc ccaaccctgg accaagcaca tcacagccgc agagcagcat gggatactca 2281 gctacctccc agcagcctcc acagtactca catcagacac atcggtactg aagatccggt 2341 accgtcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt 2401 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact 2461 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat 2521 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc 2581 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc 2641 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt 2701 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc 2761 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct 2821 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat 2881 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc 2941 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc 3001 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg 3061 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa 3121 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 3181 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 3241 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca 3301 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 3361 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct 3421 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgatga 3481 aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg 3541 tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag 3601 gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt 3661 atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg 3721 aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag 3781 acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc atggatgcga 3841 tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg 3901 gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact 3961 ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga 4021 tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca 4081 acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt 4141 tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta 4201 tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc 4261 tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca 4321 atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg 4381 ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg 4441 tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat 4501 agcacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa 4561 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct 4621 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca 4681 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca 4741 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat 4801 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag 4861 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg 4921 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa 4981 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca 5041 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 5101 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 5161 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 5221 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 5281 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 5341 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 5401 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 5461 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 5521 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 5581 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 5641 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 5701 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 5761 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 5821 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa 5881 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 5941 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga 6001 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac 6061 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg 6121 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg 6181 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt 6241 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct 6301 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat 6361 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta 6421 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca 6481 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat 6541 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac 6601 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa 6661 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt 6721 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg 6781 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat 6841 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt 6901 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc //