LOCUS Exported 6627 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCMV6M-Pak1 K299R SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6627) AUTHORS Sells MA, Knaus UG, Bagrodia S, Ambrose DM, Bokoch GM, Chernoff J TITLE Human p21-activated kinase (Pak1) regulates actin organization in mammalian cells. JOURNAL Curr Biol. 1997 Mar 1. 7(3):202-10. PUBMED 9395435 REFERENCE 2 (bases 1 to 6627) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6627 /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" CDS 656..685 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS complement(1230..1247) /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(2850..2869) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" primer_bind complement(3297..3317) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3302..3631 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3482..3617 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3544..3563 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(3669..3688) /label=T7 /note="T7 promoter, forward primer" promoter complement(3670..3688) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3702..3718) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3702..3718) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3715..3737) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3726..3742 /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(3750..3780) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3795..3816 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3933..3950) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4104..4692) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4184..4203) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4863..5723) /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 5486..5505 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5724..5828) /gene="bla" /label=AmpR promoter rep_origin 5855..6310 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5942..5961) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6152..6173 /label=F1ori-F /note="F1 origin, forward primer" primer_bind 6436..6458 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 6450..6467 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 6451..6467 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" 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 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc 421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa 541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc 601 agaattaatt cagatctggt accacgcgta tcgataagct tcgatcgtcg acatggaaca 661 gaaactcatc tctgaagagg atctgtctag aggatccatg tcaaataacg gcctagacat 721 tcaagacaaa cccccagccc ctccgatgag aaataccagc actatgattg gagccggcag 781 caaagatgct ggaaccctaa accatggttc taaacctctg cctccaaacc cagaggagaa 841 gaaaaagaag gaccgatttt accgatccat tttacctgga gataaaacaa ataaaaagaa 901 agagaaagag cggccagaga tttctctccc ttcagatttt gaacacacaa ttcatgtcgg 961 ttttgatgct gtcacagggg agtttacggg aatgccagag cagtgggccc gcttgcttca 1021 gacatcaaat atcactaagt cggagcagaa gaaaaacccg caggctgttc tggatgtgtt 1081 ggagttttac aactcgaaga agacatccaa cagccagaaa tacatgagct ttacagataa 1141 gtcagctgag gattacaatt cttctaatgc cttgaatgtg aaggctgtgt ctgagactcc 1201 tgcagtgcca ccagtttcag aagatgagga tgatgatgat gatgatgcta ccccaccacc 1261 agtgattgct ccacgcccag agcacacaaa atctgtatac acacggtctg tgattgaacc 1321 acttcctgtc actccaactc gggacgtggc tacatctccc atttcaccta ctgaaaataa 1381 caccactcca ccagatgctt tgacccggaa tactgagaag cagaagaaga agcctaaaat 1441 gtctgatgag gagatcttgg agaaattacg aagcatagtg agtgtgggcg atcctaagaa 1501 gaaatataca cggtttgaga agattggaca aggtgcttca ggcaccgtgt acacagcaat 1561 ggatgtggcc acaggacagg aggtcgcgat taggcagatg aatcttcagc agcagcccaa 1621 gaaagagctg attattaatg agatcctggt catgagggaa aacaagaacc caaacattgt 1681 gaattacttg gacagttacc tcgtgggaga tgagctgtgg gttgttatgg aatacttggc 1741 tggaggctcc ttgacagatg tggtgacaga aacttgcatg gatgaaggcc aaattgcagc 1801 tgtgtgccgt gagtgtctgc aggctctgga gttcttgcat tcgaaccagg tcattcacag 1861 agacatcaag agtgacaata ttctgttggg aatggatggc tctgtcaagc taactgactt 1921 tggattctgt gcacagataa ccccagagca gagcaaacgg agcaccatgg taggaacccc 1981 atactggatg gcaccagagg ttgtgacacg aaaggcctat gggcccaagg ttgacatctg 2041 gtccctgggc atcatggcca tcgaaatgat tgaaggggag cctccatacc tcaatgaaaa 2101 ccctctgaga gccttgtacc tcattgccac caatgggacc ccagaacttc agaacccaga 2161 gaagctgtca gctatcttcc gggactttct gaaccgctgt ctcgagatgg atgtggagaa 2221 gagaggttca gctaaagagc tgatacagca tcaattcctg aagattgcca agcccctctc 2281 cagcctcact ccactgattg ctgcagctaa ggaggcaaca aagaacaatc actaaaacca 2341 cactcacccc agcctcattg tgccaagcct tctgtgagat aaatgcacat ttcagaaatt 2401 ccaactcctg atgccctctt ctccttgcct tgcttctccc atttcctgat ctagcactcc 2461 tcaagacttt gatccttgga aaccgtgtgt ccagcattga agagaactgc aactgaatga 2521 ctaatcagat gatggccatt tctaaataag gaatttcctc ccaattcatg gatatgaggg 2581 tggtttatga ttaagggttt atataaataa atgtttctag tcttccgggc cttcgtggcc 2641 tcgaggaatt ccgtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt 2701 tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga 2761 ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt 2821 tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc 2881 acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc 2941 tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg 3001 gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat 3061 ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc 3121 tgtccttctg attttaaaat aactatacca gcaggaggac gtccagacac agcataggct 3181 acctggccat gcccaaccgg tgggacattt gagttgcttg cttggcactg tcctctcatg 3241 cgttgggtcc actcagtaga tgcctgttga attgggtacg cggccagctt ggctgtggaa 3301 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag 3361 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag 3421 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc 3481 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt 3541 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg 3601 aggctttttt ggaggcctag gcttttgcaa aaagctcctc gaggaactga aaaaccagaa 3661 agttaattcc ctatagtgag tcgtattaaa ttcgtaatca tgtcatagct gtttcctgtg 3721 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa 3781 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct 3841 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga 3901 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc 3961 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 4021 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 4081 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 4141 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 4201 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 4261 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 4321 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 4381 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 4441 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 4501 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc 4561 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 4621 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 4681 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 4741 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 4801 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 4861 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc 4921 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc 4981 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata 5041 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc 5101 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc 5161 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca 5221 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa 5281 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca 5341 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt 5401 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt 5461 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg 5521 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga 5581 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc 5641 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg 5701 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag 5761 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg 5821 gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc 5881 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc 5941 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct 6001 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa 6061 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc 6121 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca 6181 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat 6241 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg 6301 cttacaattt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg 6361 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg 6421 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct 6481 atacattgaa tcaatattgg caattagcca tattagtcat tggttatata gcataaatca 6541 atattggcta ttggccattg catacgttgt atctatatca taatatgtac atttatattg 6601 gctcatgtcc aatatgaccg ccatgtt //