LOCUS Exported 6177 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION for generation of AID strains using CRISPR tagging technology. ACCESSION . VERSION . KEYWORDS pLinker-AID-Ty-HXGPRT-LoxP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6177) AUTHORS Long S, Brown KM, Drewry LL, Anthony B, Phan IQH, Sibley LD TITLE Calmodulin-like proteins localized to the conoid regulate motility and cell invasion by Toxoplasma gondii. JOURNAL PLoS Pathog. 2017 May 5;13(5):e1006379. doi: 10.1371/journal.ppat.1006379. PUBMED 28475612 REFERENCE 2 (bases 1 to 6177) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6177 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 7..40 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind 1977..2010 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." primer_bind complement(2054..2073) /label=T7 /note="T7 promoter, forward primer" promoter complement(2055..2073) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2080..2097) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2080..2096) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2089..2111) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 2238..2693 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2325..2344) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2535..2556 /label=F1ori-F /note="F1 origin, forward primer" promoter 2719..2823 /gene="bla" /label=AmpR promoter CDS 2824..3684 /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 complement(3042..3061) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3855..4443 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4344..4363 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4597..4614 /label=L4440 /note="L4440 vector, forward primer" protein_bind 4731..4752 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4767..4797 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4805..4821 /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)." primer_bind 4810..4832 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 4829..4845 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 4829..4845 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 4866..4884 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" CDS 4989..5672 /codon_start=1 /gene="Arabidopsis thaliana AXR3" /product="auxin-responsive IAA17 protein" /label=auxin-inducible degron /note="acts as an auxin-inducible degron (AID) in the presence of TIR1" /translation="MGSVELNLRETELCLGLPGGDTVAPVTGNKRGFSETVDLKLNLNN EPANKEGSTTHDVVTFDSKEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSG GPEAAAFVKVSMDGAPYLRKIDLRMYKSYDELSNALSNMFSSFTMGKHGGEEGMIDFMN ERKLMDLVNSWDYVPSYEDKDGDWMLVGDVPWPMFVDTCKRLRLMKGSDAIGLAPRAME KCKSRA" primer_bind 6158..6174 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 6159..6175 /label=KS primer /note="common sequencing primer, one of multiple similar variants" ORIGIN 1 aagcttataa cttcgtatag catacattat acgaagttat cagcacgaaa ccttgcattc 61 aaacccgccc gcggaagatc cgatcttgct gctgttcgca gtcccagtag cgtcctgtcg 121 gccgcgccgt ctctgttggt gggcagccgc tacacctgtt atctgactgc cgtgcgcgaa 181 aatgacgcca tttttgggaa aatcggggaa cttcattctt taaaagtatg cggaggtttc 241 ctttttcttc tgttcgtttc tttttctcgg gtttgataac cgtgttcgat gtaagcactt 301 tccgtctctc ctccgtgctt tgttcgacat cgagaccagg tgtgcagatc cttcgcttgt 361 cgatccggag acgcgtgtct cgtagaacct tttcatttta ccacacggca gtgcggagca 421 ctgctctgag tgcagcaggg acgggtgaag tttcgcttta gtagtgcgtt tctgctctac 481 ggggcgttgt cagatccagc aaaatggcgt ccaaacccat tgaagactac ggcaagggca 541 agggccgtat tgagcccatg tatatccccg acaacacctt ctacaacgct gatgactttc 601 ttgtgccccc ccactgcaag ccctacattg acaaaatcct cctccctggt ggattggtca 661 aggacagagt tgagaagttg gcgtatgaca tccacagaac ttacttcggc gaggagttgc 721 acatcatttg catcctgaaa ggctctcgcg gcttcttcaa ccttctgatc gactaccttg 781 ccaccataca gaagtacagt ggtcgtgagt ccagcgtgcc ccccttcttc gagcactatg 841 tccgcctgaa gtcctaccag aacgacaaca gcacaggcca gctcaccgtc ttgagcgacg 901 acttgtcaat ctttcgcgac aagcacgttc tgattgttga ggacatcgtc gacaccggtt 961 tcaccctcac cgagttcggt gagcgcctga aagccgtcgg tcccaagtcg atgagaatcg 1021 ccaccctcgt cgagaagcgc acagatcgct ccaacagctt gaagggcgac ttcgtcggct 1081 tcagcattga agacgtctgg atcgttggtt gctgctacga cttcaacgag atgttccgcg 1141 acttcgacca cgtcgccgtc ctgagcgacg ccgctcgcaa aaagttcgag aagtaaaccc 1201 tgcatagccc acagaagctg cccgtctctc gttttcctct cttttcggag ggatcaggga 1261 gagtgcctcg gatcggagag agctgacgag ggggtgccag agacccctgt gtcctttatc 1321 gaagaaaagg gatgactctt catgtggcat ttcacacagt ctcacctcgc cttgttttct 1381 ttttgtcaat cagaacgaaa gcgagttgcg ggtgacgcag atgtgcgtgt atccactcgt 1441 gaatgcgtta tcgttctgta tgccgctaga gtgctggact gttgctgtct gcccacgaca 1501 gcagacaact ttccttctat gcacttgcag gatggtgcag cgcaaacgac ggagagaaag 1561 gagcaccctc tcagtttccc tacgatgtgc tgtcagtttc gactcttcac cgcgaacgat 1621 tggcgatacg tctctgttga cttgttaggc tccgaccacg aagctccctt aactagataa 1681 gccgcgacac ctaagtgtac accatttgca gatcgataat ctgcgaccgc tgaatccgtc 1741 cagatcagta aaaccgcacc acctaagtgt aaaccttgtt taggtcgata aaatgctacc 1801 aacccccacc cacaatcgag ccttgagcgt ttctgcgcac gcgttggcct acgtgacttg 1861 ctgatgcctg cctctggcca ttcatgccag tcagtgcgca taaaaatgtg gacacagtcg 1921 gttgacaagt gttctggcag gctacagtga caccgcggtg gagggggatc attgaaataa 1981 cttcgtatag catacattat acgaagttat ctgcagtcta gagcggccgc caccgcggtg 2041 gagctccaat tcgccctata gtgagtcgta ttacaattca ctggccgtcg ttttacaacg 2101 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt 2161 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag 2221 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt 2281 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt 2341 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc 2401 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga 2461 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc 2521 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt 2581 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct 2641 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc 2701 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat 2761 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag 2821 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt 2881 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt 2941 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc 3001 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta 3061 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac 3121 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa 3181 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg 3241 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc 3301 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg 3361 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta 3421 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg 3481 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg 3541 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc 3601 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt 3661 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt 3721 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc 3781 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag 3841 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa 3901 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg 3961 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag 4021 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg 4081 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga 4141 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc 4201 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc 4261 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga 4321 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt 4381 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg 4441 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac 4501 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga 4561 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg 4621 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc 4681 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt 4741 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt 4801 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag 4861 ctcgaaatta accctcacta aagggaacaa aagctggtac cctgtacttc caatccaatt 4921 taattaaaat tggaagtgga ggacgggaat tcgggcccgc tagcaagggc tcgggctcga 4981 cccagctgat gggcagtgtc gagctgaatc tgagggagac tgagctgtgt cttggtcttc 5041 ccggtggaga tacagtggct ccggtaaccg gaaacaagag agggttctca gagacggttg 5101 atctgaagct aaatctgaat aatgagcctg caaacaagga aggatctacg actcatgacg 5161 tcgtgacttt tgattccaag gagaagagtg cttgtcctaa agatccagcc aaacctccgg 5221 ccaaggcaca agttgtggga tggccaccgg tgagatcata ccggaagaac gtgatggttt 5281 cctgccaaaa atcaagcggt ggcccggagg cggcggcgtt cgtgaaggta tcaatggacg 5341 gagcaccgta cttgaggaaa atcgatttga ggatgtataa aagctacgat gagctttcta 5401 atgctttgtc caacatgttc agctctttta ccatgggcaa acatggagga gaagaaggaa 5461 tgatagactt catgaatgag aggaaattga tggatttggt gaatagctgg gactatgttc 5521 cctcttatga agacaaagac ggtgattgga tgctcgtcgg cgacgttcct tggccaatgt 5581 tcgtcgatac atgcaagcgt ttacgtctca tgaaaggatc ggatgccatt ggtctcgctc 5641 cgagggcgat ggagaagtgc aagagcagag ctgggcgcgc cggctctggc ggcgaggtcc 5701 acacgaacca ggacccgctc gattaacccg ggcatatgta gaaaagttgt aacgttagta 5761 aacgtaacac acgggtataa tagccattta atggcgccgt atatttttca ctccccctct 5821 ccgataggag gaagaaagac gatccgtctg cgaatgtata ttacatcgac cggccatagt 5881 acgagaagtg gtgccggtat tgttctgtcc ggggggtgta ccatcctatt ttttgtatat 5941 gtcgttcttg tcggggagca acagcccatc ttccggctat tgtctcgtat cagttgtgaa 6001 acagactgcg ttttgccgaa aatgagtcct atgtccgatg gctatttgta ccgcttcgta 6061 ccatttcgct tgtcagctgt aatcaaccga attcatttgg cgagacatga ttccgattaa 6121 attctttact gcgcgtgctc ggatccacta gttctactcg aggtcgacgg tatcgat //