LOCUS Exported 6609 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS HA-Cdk9 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6609) AUTHORS Wang Y, Dow EC, Liang YY, Ramakrishnan R, Liu H, Sung TL, Lin X, Rice AP TITLE Phosphatase PPM1A regulates phosphorylation of Thr-186 in the Cdk9 T-loop. JOURNAL J Biol Chem. 2008 Nov 28. 283(48):33578-84. PUBMED 18829461 REFERENCE 2 (bases 1 to 6609) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6609 /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 898..978 /codon_start=1 /product="three tandem HA epitope tags" /label=3xHA /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA" promoter complement(2161..2179) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2162..2179) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(2199..2216) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2205..2429 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2475..2903 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2562..2581) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2772..2793 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2912..2932) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2917..3247 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3098..3233 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3160..3179 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3314..4108 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3368..3387) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3978..3997 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4282..4403 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4319..4338) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4373..4392 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4452..4468) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4452..4468) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4465..4487) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4476..4492 /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(4500..4530) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4545..4566 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4683..4700) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4854..5442) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4934..4953) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5613..6473) /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 6236..6255 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6474..6578) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttatgtac 901 ccatacgacg tcccagacta cgcttaccca tacgacgtcc cagactacgc ttacccatac 961 gacgtcccag actacgctac cggtgcaaag cagtacgact cggtggagtg ccctttttgt 1021 gatgaagttt ccaaatacga gaagctcgcc aagatcggcc aaggcacctt cggggaggtg 1081 ttcaaggcca ggcaccgcaa gaccggccag aaggtggctc tgaagaaggt gctgatggaa 1141 aacgagaagg aggggttccc cattacagcc ttgcgggaga tcaagatcct tcagcttcta 1201 aaacacgaga atgtggtcaa cttgattgag atttgtcgaa ccaaagcttc cccctataac 1261 cgctgcaagg gtagtatata cctggtgttc gacttctgcg agcatgacct tgctgggctg 1321 ttgagcaatg ttttggtcaa gttcacgctg tctgagatca agagggtgat gcagatgctg 1381 cttaacggcc tctactacat ccacagaaac aagatcctgc atagggacat gaaggctgct 1441 aatgtgctta tcactcgtga tggggtcctg aagctggcag actttgggct ggcccgggcc 1501 ttcagcctgg ccaagaacag ccagcccaac cgctacacca accgtgtggt gacactctgg 1561 taccggcccc cggagctgtt gctcggggag cgggactacg gcccccccat tgacctgtgg 1621 ggtgctgggt gcatcatggc agagatgtgg acccgcagcc ccatcatgca gggcaacacg 1681 gagcagcacc aactcgccct catcagtcag ctctgcggct ccatcacccc tgaggtgtgg 1741 ccaaacgtgg acaactatga gctgtacgaa aagctggagc tggtcaaggg ccagaagcgg 1801 aaggtgaagg acaggctgaa ggcctatgtg cgtgacccat acgcactgga cctcatcgac 1861 aagctgctgg tgctggaccc tgcccagcgc atcgacagcg atgacgccct caaccacgac 1921 ttcttctggt ccgaccccat gccctccgac ctcaagggca tgctctccac ccacctgacg 1981 tccatgttcg agtacttggc accaccgcgc cggaagggca gccagatcac ccagcagtcc 2041 accaaccaga gtcgcaatcc cgccaccacc aaccagacgg agtttgagcg cgtcttctga 2101 gaattctgca gatatccatc acactggcgg ccgctcgagc atgcatctag agggccctat 2161 tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg 2221 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc 2281 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc 2341 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag 2401 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc 2461 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac 2521 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc 2581 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt 2641 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg 2701 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac 2761 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta 2821 ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat 2881 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag 2941 tccccaggct ccccaggcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac 3001 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa 3061 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag 3121 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc 3181 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt 3241 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg 3301 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt 3361 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt 3421 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc 3481 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc 3541 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga 3601 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat 3661 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca 3721 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga 3781 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc 3841 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat 3901 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga 3961 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg 4021 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt 4081 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa 4141 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg 4201 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg 4261 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc 4321 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 4381 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg 4441 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 4501 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 4561 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg 4621 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 4681 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 4741 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 4801 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 4861 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 4921 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 4981 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 5041 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 5101 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 5161 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 5221 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 5281 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 5341 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 5401 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 5461 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 5521 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 5581 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 5641 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 5701 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 5761 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 5821 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 5881 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 5941 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 6001 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 6061 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 6121 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 6181 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 6241 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 6301 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 6361 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 6421 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 6481 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 6541 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 6601 cctgacgtc //