LOCUS Exported 7239 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS PKC iota WT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7239) AUTHORS Soh JW, Weinstein IB TITLE Roles of specific isoforms of protein kinase C in the transcriptional control of cyclin D1 and related genes. JOURNAL J Biol Chem. 2003 Sep 5. 278(36):34709-16. PUBMED 12794082 REFERENCE 2 (bases 1 to 7239) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7239 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 381..584 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 535..555 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 629..648 /label=T7 /note="T7 promoter, forward primer" promoter 629..647 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 684..693 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2463..2489 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" promoter complement(2558..2576) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2559..2576) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(2596..2613) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2602..2826 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2872..3300 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2959..2978) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3169..3190 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3309..3329) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3314..3643 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3494..3629 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3556..3575 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3710..4504 /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(3764..3783) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4374..4393 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4678..4799 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4715..4734) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4769..4788 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4848..4864) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4848..4864) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4861..4883) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4872..4888 /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(4896..4926) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4941..4962 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5079..5096) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5250..5838) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5330..5349) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6009..6869) /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 6632..6651 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6870..6974) /gene="bla" /label=AmpR promoter primer_bind complement(7049..7068) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag 601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt 661 ggtaccgagc tcggatcctc gaggccacca tggaattctc ccacacggtc gcaggcggcg 721 gcagcgggga ccattcccac caggtccggg tgaaagccta ctaccgcggg gatatcatga 781 taacacattt tgaaccttcc atctcctttg agggcctttg caatgaggtt cgagacatgt 841 gttcttttga caacgaacag ctcttcacca tgaaatggat agatgaggaa ggagacccgt 901 gtacagtatc atctcagttg gagttagaag aagcctttag actttatgag ctaaacaagg 961 attctgaact cttgattcat gtgttccctt gtgtaccaga acgtcctggg atgccttgtc 1021 caggagaaga taaatccatc taccgtagag gtgcacgccg ctggagaaag ctttattgtg 1081 ccaatggcca cactttccaa gccaagcgtt tcaacaggcg tgctcactgt gccatctgca 1141 cagaccgaat atggggactt ggacgccaag gatataagtg catcaactgc aaactcttgg 1201 ttcataagaa gtgccataaa ctcgtcacaa ttgaatgtgg gcggcattct ttgccacagg 1261 aaccagtgat gcccatggat cagtcatcca tgcattctga ccatgcacag acagtaattc 1321 catataatcc ttcaagtcat gagagtttgg atcaagttgg tgaagaaaaa gaggcaatga 1381 acaccaggga aagtggcaaa gcttcatcca gtctaggtct tcaggatttt gatttgctcc 1441 gggtaatagg aagaggaagt tatgccaaag tactgttggt tcgattaaaa aaaacagatc 1501 gtatttatgc aatgaaagtt gtgaaaaaag agcttgttaa tgatgatgag gatattgatt 1561 gggtacagac agagaagcat gtgtttgagc aggcatccaa tcatcctttc cttgttgggc 1621 tgcattcttg ctttcagaca gaaagcagat tgttctttgt tatagagtat gtaaatggag 1681 gagacctaat gtttcatatg cagcgacaaa gaaaacttcc tgaagaacat gccagatttt 1741 actctgcaga aatcagtcta gcattaaatt atcttcatga gcgagggata atttatagag 1801 atttgaaact ggacaatgta ttactggact ctgaaggcca cattaaactc actgactacg 1861 gcatgtgtaa ggaaggatta cggccaggag atacaaccag cactttctgt ggtactccta 1921 attacattgc tcctgaaatt ttaagaggag aagattatgg tttcagtgtt gactggtggg 1981 ctcttggagt gctcatgttt gagatgatgg caggaaggtc tccatttgat attgttggga 2041 gctccgataa ccctgaccag aacacagagg attatctctt ccaagttatt ttggaaaaac 2101 aaattcgcat accacgttct ctgtctgtaa aagctgcaag tgttctgaag agttttctta 2161 ataaggaccc taaggaacga ttgggttgtc atcctcaaac aggatttgct gatattcagg 2221 gacacccgtt cttccgaaat gttgattggg atatgatgga gcaaaaacag gtggtacctc 2281 cctttaaacc aaatatttct ggggaatttg gtttggacaa ctttgattct cagtttacta 2341 atgaacctgt ccagctcact ccagatgacg atgacattgt gaggaagatt gatcagtctg 2401 aatttgaagg ttttgagtat atcaatcctc ttttgatgtc tgcagaagaa tgtgtcgaat 2461 tctatcctta cgacgtgcct gactacgcct aaggatccaa ttctgcagat atccatcaca 2521 ctggcggccg ctcgagcatg catctagagg gccctattct atagtgtcac ctaaatgcta 2581 gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct 2641 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg 2701 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc 2761 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct 2821 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct 2881 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg 2941 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg 3001 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac 3061 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct 3121 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt 3181 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt 3241 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt 3301 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag 3361 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc 3421 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc 3481 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg 3541 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca 3601 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg 3661 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca 3721 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg 3781 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg 3841 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc 3901 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt 3961 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc 4021 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca 4081 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc 4141 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg 4201 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct 4261 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc 4321 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc 4381 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta 4441 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt 4501 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga 4561 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac 4621 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac 4681 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat 4741 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat 4801 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt 4861 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag 4921 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg 4981 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg 5041 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc 5101 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc 5161 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg 5221 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 5281 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 5341 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 5401 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 5461 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 5521 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 5581 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 5641 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt 5701 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 5761 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 5821 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 5881 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 5941 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 6001 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 6061 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 6121 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 6181 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 6241 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 6301 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 6361 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 6421 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 6481 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 6541 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 6601 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 6661 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 6721 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 6781 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 6841 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 6901 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 6961 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat 7021 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc 7081 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag 7141 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt 7201 ttgcgctgct tcgcgatgta cgggccagat atacgcgtt //