LOCUS Exported 8153 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS HA.PKD SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8153) AUTHORS Storz P, Toker A TITLE Protein kinase D mediates a stress-induced NF-kappaB activation and survival pathway. JOURNAL EMBO J. 2003 Jan 2. 22(1):109-20. PUBMED 12505989 REFERENCE 2 (bases 1 to 8153) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8153 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(11..28) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(182..770) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(262..281) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(941..1801) /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 1564..1583 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1802..1906) /gene="bla" /label=AmpR promoter primer_bind complement(1981..2000) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 2172..2551 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2552..2755 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2706..2726 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 2800..2819 /label=T7 /note="T7 promoter, forward primer" promoter 2800..2818 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter complement(5643..5661) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(5644..5661) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(5681..5698) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5687..5911 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 5957..6385 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6044..6063) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6254..6275 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(6394..6414) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 6399..6728 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 6579..6714 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6641..6660 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 6795..7589 /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(6849..6868) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 7459..7478 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 7763..7884 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(7800..7819) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 7854..7873 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(7933..7949) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7933..7949) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7946..7968) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7957..7973 /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(7981..8011) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 8026..8047 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." ORIGIN 1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 1921 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag 1981 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga 2041 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa 2101 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag 2161 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt 2221 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg 2281 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac 2341 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt 2401 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa 2461 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta 2521 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg 2581 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg 2641 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc 2701 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg 2761 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga 2821 gacccaagct tggtaccgag ctcggatcca tgtatcctta tgatgttctt gattatgcta 2881 gcgcccctcc ggtcctgcgg ccgcccagtc cgctgctgcc cgtggcggcg gcagctgccg 2941 cagcggccgc cgcactggtc ccagggtccg ggcccgggcc cgcgccgttc ttggctcctg 3001 tcgcggcccc ggtcgggggc atctcgttcc atctgcagat cggcctgagc cgtgagccgg 3061 tgctgctgct gcaggactcg tccggggact acagcctggc gcacgtccgc gagatggctt 3121 gctccattgt cgaccagaag ttccctgaat gtggtttcta cggaatgtat gataagatcc 3181 tgctttttcg ccatgaccct acctctgaaa acatccttca gctggtgaaa gcggccagtg 3241 atatccagga aggcgatctt attgaagtgg tcttgtcagc ttccgccacc tttgaagact 3301 ttcagattcg tccccacgct ctctttgttc attcatacag agctccagct ttctgtgatc 3361 actgtggaga aatgctgtgg gggctggtac gtcaaggtct taaatgtgaa gggtgtggtc 3421 tgaattacca taagagatgt gcatttaaaa tacccaacaa ttgcagcggt gtgaggcgga 3481 gaaggctctc aaacgtttcc ctcactgggg tcagcaccat ccgcacatca tctgctgaac 3541 tctctacaag tgcccctgat gagccccttc tgcaaaaatc accatcagag tcgtttattg 3601 gtcgagagaa gaggtcaaat tctcaatcat acattggacg accaattcac cttgacaaga 3661 ttttgatgtc taaagttaaa gtgccgcaca catttgtcat ccactcctac acccggccca 3721 cagtgtgcca gtactgcaag aagcttctga aggggctttt caggcagggc ttgcagtgca 3781 aagattgcag attcaactgc cataaacgtt gtgcaccgaa agtaccaaac aactgccttg 3841 gcgaagtgac cattaatgga gatttgctta gccctggggc agagtctgat gtggtcatgg 3901 aagaagggag tgatgacaat gatagtgaaa ggaacagtgg gctcatggat gatatggaag 3961 aagcaatggt ccaagatgca gagatggcaa tggcagagtg ccagaacgac agtggcgaga 4021 tgcaagatcc agacccagac cacgaggacg ccaacagaac catcagtcca tcaacaagca 4081 acaatatccc actcatgagg gtagtgcagt ctgtcaaaca cacgaagagg aaaagcagca 4141 cagtcatgaa agaaggatgg atggtccact acaccagcaa ggacacgctg cggaaacggc 4201 actattggag attggatagc aaatgtatta ccctctttca gaatgacaca ggaagcaggt 4261 actacaagga aattccttta tctgaaattt tgtctctgga accagtaaaa acttcagctt 4321 taattcctaa tggggccaat cctcattgtt tcgaaatcac tacggcaaat gtagtgtatt 4381 atgtgggaga aaatgtggtc aatccttcca gcccatcacc aaataacagt gttcccacca 4441 gtggcgttgg tgcagatgtg gccaggatgt gggagatagc catccagcat gcccttatgc 4501 ccgtcattcc caagggctcc tccgtgggta caggaaccaa cttgcacaga gatatctctg 4561 tgagtatttc agtatcaaat tgccagattc aagaaaatgt ggacatcagc acagtatatc 4621 agatttttcc tgatgaagta ctgggttctg gacagtttgg aattgtttat ggaggaaaac 4681 atcgtaaaac aggaagagat gtagctatta aaatcattga caaattacga tttccaacaa 4741 aacaagaaag ccagcttcgt aatgaggttg caattctaca gaaccttcat caccctggtg 4801 ttgtaaattt ggagtgtatg tttgagacgc ctgaaagagt gtttgttgtt atggaaaaac 4861 tccatggaga catgctggaa atgatcttgt caagtgaaaa gggcaggttg ccagagcaca 4921 taacgaagtt tttaattact cagatactcg tggctttgcg gcaccttcat tttaaaaata 4981 tcgttcactg tgacctcaaa ccagaaaatg tgttgctagc ctcagctgat ccttttcctc 5041 aggtgaaact ttgtgatttt ggttttgccc ggatcattgg agagaagtct ttccggaggt 5101 cagtggtggg tacccccgct tacctggctc ctgaggtcct aaggaacaag ggctacaatc 5161 gctctctaga catgtggtct gttggggtca tcatctatgt aagcctaagc ggcacattcc 5221 catttaatga agatgaagac atacacgacc aaattcagaa tgcagctttc atgtatccac 5281 caaatccctg gaaggaaata tctcatgaag ccattgatct tatcaacaat ttgctgcaag 5341 taaaaatgag aaagcgctac agtgtggata agaccttgag ccacccttgg ctacaggact 5401 atcagacctg gttagatttg cgagagctgg aatgcaaaat cggggagcgc tacatcaccc 5461 atgaaagtga tgacctgagg tgggagaagt atgcaggcga gcagcggctg cagtacccca 5521 cacacctgat caatccaagt gctagccaca gtgacactcc tgagactgaa gaaacagaaa 5581 tgaaagccct cggtgagcgt gtcagcatcc tctgactcga gcatgcatct agagggccct 5641 attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt gccttctagt 5701 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact 5761 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat 5821 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc 5881 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc 5941 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt 6001 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc 6061 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct 6121 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat 6181 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc 6241 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc 6301 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg 6361 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa 6421 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 6481 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 6541 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca 6601 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 6661 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct 6721 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat 6781 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg 6841 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg 6901 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg 6961 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc 7021 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg 7081 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca 7141 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc 7201 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg 7261 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg 7321 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata 7381 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg 7441 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat 7501 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct 7561 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca 7621 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt 7681 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat 7741 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag 7801 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt 7861 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt 7921 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 7981 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 8041 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 8101 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gct //