LOCUS Exported 4835 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION integration of loxP-ura4+-loxM3 cassette. ACCESSION . VERSION . KEYWORDS pAW1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4835) AUTHORS Watson AT1, Garcia V, Bone N, Carr AM, Armstrong J. TITLE Gene tagging and gene replacement using recombinase-mediated cassette exchange in Schizosaccharomyces pombe. JOURNAL Gene. 2008 Jan 15;407(1-2):63-74 PUBMED 18054176 REFERENCE 2 (bases 1 to 4835) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4835 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 22..55 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS 588..1382 /codon_start=1 /gene="Schizosaccharomyces pombe ura4" /product="orotidine 5'-phosphate decarboxylase, required for uracil biosynthesis" /label=S. pombe ura4 /translation="MDARVFQSYSARAEGMKNPIAKELLALMEEKQSNLSVAVDLTKKS EILELVDKIGPYVCVIKTHIDVVEDFDQDMVEKLVALGKKHRFLIFEDRKFADIGNTVK LQYASGVYKIASWAHITNCHTVSGEGIIQGLKEVGLPLGRGLLLLAEMSSKGSLATGSY TEKTLEWFEKHTDFCFGFIAGRRFPNLQSDYITMSPGIGLDVKGDGLGQQYRTPEEVIV NCGSDIIIVGRGVYGAGRNPVVEAKRYREAGWKAYQQRLSQH" primer_bind complement(1886..1902) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1887..1903) /label=pBluescriptKS /note="For pBluescript vector" primer_bind complement(1932..1952) /label=T3 /note="T3 promoter, forward primer" promoter complement(1932..1950) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(1971..1987) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1971..1987) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1984..2006) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1995..2011 /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(2019..2049) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2064..2085 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2202..2219) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2373..2961) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2453..2472) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3132..3992) /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 3755..3774 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3993..4097) /gene="bla" /label=AmpR promoter rep_origin complement(4123..4578) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4260..4281) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4472..4491 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4705..4727 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 4719..4736 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 4720..4736 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 4746..4765 /label=T7 /note="T7 promoter, forward primer" promoter 4746..4764 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind 4797..4813 /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind 4797..4813 /label=pBluescriptSK /note="For pBluescript vector" ORIGIN 1 cggatccccg ggttaattaa cataacttcg tatagcatac attatacgaa gttatagctt 61 agctacaaat cccactggct atatgtatgc atttgtgtta aaaaagtttg tatagattat 121 ttaatctact cagcattctt tctctaaata ggaatttgtt acttaatgga gaaaaaaatg 181 tttcgattta cctagtgtat ttgtttgtat actcacgttt aatttcaaac atccattcta 241 tcttgtgtaa tttttggcat ggtgaaaaag ataatcagcc ttataatctt tacaaaagta 301 agaaattctg taaataagcc ttaatgccct tgctttaaat taaaatggtt ctttttcatg 361 ataatgtttg cactttgtga atatatttta gatagttctg tgaggtataa ttaagatgtt 421 ttagagactt atacaatttt gtctttataa attcttaatt gattttacca tcccagttta 481 actatgcttc gtcggcatct ctgcacatgt cgtgttttct taccgtattg tcctaccaag 541 aacctctttt ttgcttggat cgaaattaaa ggtttaaaag caaagttatg gatgctagag 601 tatttcaaag ctattcagct agagctgagg ggatgaaaaa tcccattgcc aaggaattgt 661 tggctttgat ggaagaaaag caaagcaact tgtcagtcgc ggtcgatttg acgaagaaat 721 ccgaaatctt agaattggta gataaaattg gaccctatgt ctgtgttatc aagacacata 781 ttgacgttgt cgaggatttc gaccaggata tggtagaaaa actggtggcc ttaggtaaaa 841 agcatcgttt tcttatcttt gaggatcgca aattcgcaga cattggaaat accgtcaagc 901 tacaatatgc atctggtgtg tacaaaattg cttcttgggc tcatatcaca aattgccata 961 cagtgtcagg cgagggtatt atacaaggcc tcaaagaagt tggtttacct ttgggacgtg 1021 gtctcttgct tttggctgaa atgtcttcca aaggctcttt ggctactggt tcctacacag 1081 agaaaacctt agaatggttt gagaagcata ccgatttttg ctttggcttt atagctggtc 1141 gtcgatttcc taaccttcaa agcgactaca taactatgtc ccctggtatc ggcttggatg 1201 ttaaaggaga cgggctggga cagcaatatc gtactcctga agaagtgatt gtaaactgcg 1261 gtagcgatat catcattgtt ggtcgtggag tctatggagc tggtcgtaat cctgttgtcg 1321 aagccaagag atatagagaa gctggttgga aggcatatca gcaaagactt tctcagcatt 1381 aaaaaaagac taatgtaaaa tttttttggt tggttattga aaaagtcgat gccttgtttg 1441 cgtttgtttt cctaggcgtt ttatgtcaga aggcatttag aattagtata caagtactct 1501 ttggtaaaat tttatgtagc gactaaaata ttaactatta tagataaaca ccttgggaat 1561 aaaaagtaat ttgctatagt aatttattaa acatgctcct acaacattac cacaatcttt 1621 tctcttggat tgacattgaa taagaaaaga gtgaattttt ttagacttgt aatgataact 1681 atgtacaaag ccaatgaaag atgtatgtag atgaatgtaa aataccatgt agacaaacaa 1741 gataaaactt ggttataaac attggtgttg gaacagaata aattagatgt caaaaagttt 1801 cgtcaatatc acaagctata acttcgtata tggtattata tacgaagtta tgtttaaacg 1861 agctcgaatt catcatcaag cttatcgata ccgtcgacct cgaggggggg cccggtaccc 1921 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg 1981 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cgggagcata 2041 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 2101 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 2161 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 2221 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 2281 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 2341 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 2401 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 2461 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 2521 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 2581 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 2641 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 2701 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 2761 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta 2821 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 2881 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 2941 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 3001 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 3061 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 3121 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 3181 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 3241 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta 3301 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 3361 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 3421 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt 3481 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 3541 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 3601 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 3661 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 3721 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact 3781 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg 3841 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 3901 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 3961 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 4021 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 4081 caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata 4141 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg 4201 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc 4261 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa 4321 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt 4381 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac 4441 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta 4501 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg 4561 cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat 4621 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat 4681 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagc 4741 gcgcgtaata cgactcacta tagggcgaat tggagctcca ccgcggtggc ggccgctcta 4801 gaactagtgg atcccccggg ctgcaggaat tcgat //