LOCUS Exported 6794 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS HA GSK3 beta K85A pcDNA3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6794) TITLE GSK3 plasmids REFERENCE 2 (bases 1 to 6794) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6794 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 60..439 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 440..643 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 594..614 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 688..707 /label=T7 /note="T7 promoter, forward primer" promoter 688..706 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 1627..1638 /codon_start=1 /product="WELQut protease recognition and cleavage site" /label=WELQut site /translation="WELQ" CDS 2073..2099 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" promoter complement(2172..2190) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2173..2190) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(2210..2227) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2216..2440 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2486..2914 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2573..2592) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2783..2804 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2923..2943) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2928..3257 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3108..3243 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3170..3189 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3324..4118 /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(3378..3397) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3988..4007 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4292..4413 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4329..4348) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4383..4402 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4462..4478) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4462..4478) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4475..4497) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4486..4502 /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(4510..4540) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4555..4576 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4693..4710) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4864..5452) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4944..4963) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5623..6483) /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 6246..6265 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6484..6588) /gene="bla" /label=AmpR promoter primer_bind complement(6663..6682) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg 61 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc 121 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa 181 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac 241 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca 301 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg 361 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt 421 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg 481 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg 541 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat 601 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga 661 acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga cccaagcttg 721 gtaccgagct cggatccact agtaacggcc gccagtgtgc tggaattccg gaggaaggaa 781 ggaaaaggtg attcgcgaag agagtgatca tgtcagggcg gcccagaacc acctcctttg 841 cggagagctg caagccggtg cagcagcctt cagcttttgg cagcatgaaa gttagcagag 901 acaaggacgg cagcaaggtg acaacagtgg tggcaactcc tgggcagggt ccagacaggc 961 cacaagaagt cagctataca gacactaaag tgattggaaa tggatcattt ggtgtggtat 1021 atcaagccaa actttgtgat tcaggagaac tggtcgccat cgcgaaagta ttgcaggaca 1081 agagatttaa gaatcgagag ctccagatca tgagaaagct agatcactgt aacatagtcc 1141 gattgcgtta tttcttctac tccagtggtg agaagaaaga tgaggtctat cttaatctgg 1201 tgctggacta tgttccggaa acagtataca gagttgccag acactatagt cgagccaaac 1261 agacgctccc tgtgatttat gtcaagttgt atatgtatca gctgttccga agtttagcct 1321 atatccattc ctttggaatc tgccatcggg atattaaacc gcagaacctc ttgttggatc 1381 ctgatactgc tgtattaaaa ctctgtgact ttggaagtgc aaagcagctg gtccgaggag 1441 aacccaatgt ttcgtatatc tgttctcggt actatagggc accagagttg atctttggag 1501 ccactgatta tacctctagt atagatgtat ggtctgctgg ctgtgtgttg gctgagctgt 1561 tactaggaca accaatattt ccaggggata gtggtgtgga tcagttggta gaaataatca 1621 aggtcctggg aactccaaca agggagcaaa tcagagaaat gaacccaaac tacacagaat 1681 ttaaattccc tcaaattaag gcacatcctt ggactaaggt cttccgaccc cgaactccac 1741 cggaggcaat tgcactgtgt agccgtctgc tggagtatac accaactgcc cgactaacac 1801 cactggaagc ttgtgcacat tcattttttg atgaattacg ggacccaaat gtcaaactac 1861 caaatgggcg agacacacct gcactcttca acttcaccac tcaagaactg tcaagtaatc 1921 cacctctggc taccatcctt attcctcctc atgctcggat tcaagcagct gcttcaaccc 1981 ccacaaatgc cacagcagcg tcagatgcta atactggaga ccgtggacag accaataatg 2041 ctgcttctgc atcagcttcc aactccacta gttacccata cgatgttcca gattacgcaa 2101 gcttgggtgg tcccaactag tacagtcccg acggatcctc tagagtcgag catgcatcta 2161 gagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg 2221 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa 2281 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt 2341 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa 2401 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc 2461 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt 2521 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc 2581 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg 2641 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat 2701 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg 2761 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct 2821 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa 2881 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag 2941 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 3001 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 3061 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa 3121 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 3181 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag 3241 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag 3301 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg 3361 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg 3421 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc 3481 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga 3541 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc 3601 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag 3661 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat 3721 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg 3781 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca 3841 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct 3901 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg 3961 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg 4021 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc 4081 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat 4141 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta 4201 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg 4261 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta 4321 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 4381 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag 4441 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 4501 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 4561 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 4621 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 4681 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 4741 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 4801 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 4861 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 4921 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 4981 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 5041 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 5101 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 5161 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 5221 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 5281 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt 5341 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 5401 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 5461 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 5521 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 5581 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 5641 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 5701 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 5761 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 5821 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 5881 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 5941 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 6001 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 6061 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 6121 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 6181 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 6241 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 6301 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 6361 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 6421 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 6481 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 6541 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 6601 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc 6661 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg 6721 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac 6781 aattgcatga agaa //