LOCUS       sequence-219789-        4784 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  integration of ura4+-loxM3 cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4784)
  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 4784)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene."; 
            date: "2008-01-15"; volume: "407(1-2)"; pages: "63-7"
COMMENT     Sequence Label: pAW12
FEATURES             Location/Qualifiers
     source          1..4784
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             534..1328
                     /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(1832..1848)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1833..1849)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1878..1898)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1878..1896)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1917..1933)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1917..1933)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1930..1952)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1941..1957
                     /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(join(1965..1971,1972..1989,1990..1995))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2010..2031
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2148..2165)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2319..2907)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2399..2418)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3078..3869,3870..3938))
                     /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     3701..3720
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3939..4043)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4069..4524)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4206..4227)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4418..4437
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4651..4673
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4665..4682
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4666..4682
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4692..4711
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4692..4710
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     4743..4759
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4743..4759
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 aagcttagct acaaatccca ctggctatat gtatgcattt gtgttaaaaa agtttgtata
       61 gattatttaa tctactcagc attctttctc taaataggaa tttgttactt aatggagaaa
      121 aaaatgtttc gatttaccta gtgtatttgt ttgtatactc acgtttaatt tcaaacatcc
      181 attctatctt gtgtaatttt tggcatggtg aaaaagataa tcagccttat aatctttaca
      241 aaagtaagaa attctgtaaa taagccttaa tgcccttgct ttaaattaaa atggttcttt
      301 ttcatgataa tgtttgcact ttgtgaatat attttagata gttctgtgag gtataattaa
      361 gatgttttag agacttatac aattttgtct ttataaattc ttaattgatt ttaccatccc
      421 agtttaacta tgcttcgtcg gcatctctgc acatgtcgtg ttttcttacc gtattgtcct
      481 accaagaacc tcttttttgc ttggatcgaa attaaaggtt taaaagcaaa gttatggatg
      541 ctagagtatt tcaaagctat tcagctagag ctgaggggat gaaaaatccc attgccaagg
      601 aattgttggc tttgatggaa gaaaagcaaa gcaacttgtc agtcgcggtc gatttgacga
      661 agaaatccga aatcttagaa ttggtagata aaattggacc ctatgtctgt gttatcaaga
      721 cacatattga cgttgtcgag gatttcgacc aggatatggt agaaaaactg gtggccttag
      781 gtaaaaagca tcgttttctt atctttgagg atcgcaaatt cgcagacatt ggaaataccg
      841 tcaagctaca atatgcatct ggtgtgtaca aaattgcttc ttgggctcat atcacaaatt
      901 gccatacagt gtcaggcgag ggtattatac aaggcctcaa agaagttggt ttacctttgg
      961 gacgtggtct cttgcttttg gctgaaatgt cttccaaagg ctctttggct actggttcct
     1021 acacagagaa aaccttagaa tggtttgaga agcataccga tttttgcttt ggctttatag
     1081 ctggtcgtcg atttcctaac cttcaaagcg actacataac tatgtcccct ggtatcggct
     1141 tggatgttaa aggagacggg ctgggacagc aatatcgtac tcctgaagaa gtgattgtaa
     1201 actgcggtag cgatatcatc attgttggtc gtggagtcta tggagctggt cgtaatcctg
     1261 ttgtcgaagc caagagatat agagaagctg gttggaaggc atatcagcaa agactttctc
     1321 agcattaaaa aaagactaat gtaaaatttt tttggttggt tattgaaaaa gtcgatgcct
     1381 tgtttgcgtt tgttttccta ggcgttttat gtcagaaggc atttagaatt agtatacaag
     1441 tactctttgg taaaatttta tgtagcgact aaaatattaa ctattataga taaacacctt
     1501 gggaataaaa agtaatttgc tatagtaatt tattaaacat gctcctacaa cattaccaca
     1561 atcttttctc ttggattgac attgaataag aaaagagtga atttttttag acttgtaatg
     1621 ataactatgt acaaagccaa tgaaagatgt atgtagatga atgtaaaata ccatgtagac
     1681 aaacaagata aaacttggtt ataaacattg gtgttggaac agaataaatt agatgtcaaa
     1741 aagtttcgtc aatatcacaa gctataactt cgtatatggt attatatacg aagttatgtt
     1801 taaacgagct cgaattcatc atcaagctta tcgataccgt cgacctcgag ggggggcccg
     1861 gtacccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
     1921 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccggg
     1981 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     2041 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     2101 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     2161 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2221 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2281 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2341 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2401 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2461 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2521 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2581 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2641 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     2701 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     2761 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2821 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     2881 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     2941 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3001 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3061 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3121 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3181 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3241 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3301 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3361 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     3421 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     3481 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     3541 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     3601 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     3661 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     3721 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     3781 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     3841 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     3901 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     3961 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4021 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg
     4081 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     4141 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     4201 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     4261 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     4321 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     4381 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     4441 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     4501 ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag
     4561 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     4621 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     4681 gtgagcgcgc gtaatacgac tcactatagg gcgaattgga gctccaccgc ggtggcggcc
     4741 gctctagaac tagtggatcc cccgggctgc aggaattcga tatc
//
