LOCUS       sequence-121986-        7326 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  complementation of yeast auxotrophic marker MET15.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7326)
  AUTHORS   Li D, Song JZ, Shan MH, Li SP, Liu W, Li H, Zhu J, Wang Y, Lin J, 
            Xie Z
  TITLE     A fluorescent tool set for yeast Atg proteins.
  JOURNAL   Autophagy. 2015;11(6):954-60. doi: 10.1080/15548627.2015.1040971.
  PUBMED    25998947
REFERENCE   2  (bases 1 to 7326)
  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: 
            "Autophagy."; date: "2015"; volume: "11(6):954-60. doi"; pages: " 
            10.1080/15548627.2015.1040971"
COMMENT     Sequence Label: BS-Met15
FEATURES             Location/Qualifiers
     source          1..7326
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2119..2469
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     CDS             2470..3804
                     /codon_start=1
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /product="O-acetylserine and O-acetylhomoserine 
                     sulfhydrylase, required for sulfur amino acid synthesis"
                     /label=MET17
                     /translation="MPSHFDTVQLHAGQENPGDNAHRSRAVPIYATTSYVFENSKHGSQ
                     LFGLEVPGYVYSRFQNPTSNVLEERIAALEGGAAALAVSSGQAAQTLAIQGLAHTGDNI
                     VSTSYLYGGTYNQFKISFKRFGIEARFVEGDNPEEFEKVFDERTKAVYLETIGNPKYNV
                     PDFEKIVAIAHKHGIPVVVDNTFGAGGYFCQPIKYGADIVTHSATKWIGGHGTTIGGII
                     VDSGKFPWKDYPEKFPQFSQPAEGYHGTIYNEAYGNLAYIVHVRTELLRDLGPLMNPFA
                     SFLLLQGVETLSLRAERHGENALKLAKWLEQSPYVSWVSYPGLASHSHHENAKKYLSNG
                     FGGVLSFGVKDLPNADKETDPFKLSGAQVVDNLKLASNLANVGDAKTLVIAPYFTTHKQ
                     LNDKEKLASGVTKDLIRVSVGIEFIDDIIADFQQSFETVFAGQKP"
     primer_bind     complement(5085..5101)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5085..5101)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5138..5158)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5138..5156)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5177..5193)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5177..5193)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5190..5212)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5201..5217
                     /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(5225..5231,5232..5249,5250..5255))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5270..5291
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5408..5425)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5579..6167)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5659..5678)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6338..7129,7130..7198))
                     /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     6961..6980
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7199..7303)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtacctc
      661 aattctcacc gctgttgtaa ctggagcttt aaggtgttat ctaaggaagg ataaaagaac
      721 ttaaacaaca acaaacaaaa aattaagtta caatgccagc attattaaaa agattattgt
      781 ttcaagtggg tcctcatcca aacgaaagaa cattcacctt atcctctgtt tcaactgatg
      841 ggcactatat ttccttgaga ccatttgtta agccaagcgg tgatgagtta tcttttcctt
      901 tcgaatgggc ctttgccggt acaaacgaaa cagttaaagc taatgatcaa ggaaacggtg
      961 tcgttactca agatttcaat ttctggttgg atacaaatgt gtacttgaac gttccaaaca
     1021 cccatcgtgg cgaagtgaac accacttgga aaaattggga ttctggttgt gtcgaggaaa
     1081 caggcgctgt ttacccattc ggtgccgaca aagaaagcgt ctctttcaga gaattgtggc
     1141 aaccagttga cccatcaaga gaagatctag tcatcgtctc accaaacaat gagaagttct
     1201 cgtcgaatgc taggtcaatt gtcctcaaag ttactgacga agcttatgat ggtttggtta
     1261 ttgttattgg tagatggatt caagggtttt tgtcccaaaa gaataataac actattgaag
     1321 gcttgaactt catcagatta cttgaaaaag attcaggtaa atctgagttc ttattaagct
     1381 acggtaagga agtaaacaaa attccacaaa gctacgaaaa tttgaagaaa ggttccactg
     1441 taaccagcaa tgggttgaac tgggaagtta ttgaatatca cgcttaataa aggagaataa
     1501 atcgttttct actttcttct gctgctataa taagcaccta tgggatctat atagtatttt
     1561 tataacgata gactttataa aagaaaatac ctaagtgaaa atttggtgaa ttttgagata
     1621 attgttggga ttccattttt aataaggcaa taatattagg tatgtagaat atactagaag
     1681 ttctcctcga ggatttagga atccataaaa gggaatctgc aattctacac aattctataa
     1741 atattattat catcgtttta tatgttaata ttcattgatc ctattacatt atcaatcctt
     1801 gcgtttcagc ttccactaat ttagatgact atttctcatc atttgcgtca tcttctaaca
     1861 ccgtatatga taatatacta gtaacgtaaa tactagttag tagatgatag ttgattttta
     1921 ttccaacact aagaaataat ttcgccattt cttgaatgta tttaaagata tttaatgcta
     1981 taatagacat ttaaatccaa ttcttccaac atacaatggg agtttggccg agtggtttaa
     2041 ggcgtcagat ttaggtggat ttaacctcta aaatctctga tatcttcgga tgcaagggtt
     2101 cgaatccctt agctctcatt attttttgct ttttctcttg aggtcacatg atcgcaaaat
     2161 ggcaaatggc acgtgaagct gtcgatattg gggaactgtg gtggttggca aatgactaat
     2221 taagttagtc aaggcgccat cctcatgaaa actgtgtaac ataataaccg aagtgtcgaa
     2281 aaggtggcac cttgtccaat tgaacacgct cgatgaaaaa aataagatat atataaggtt
     2341 aagtaaagcg tctgttagaa aggaagtttt tcctttttct tgctctcttg tcttttcatc
     2401 tactatttcc ttcgtgtaat acagggtcgt cagatacata gatacaattc tattaccccc
     2461 atccatacaa tgccatctca tttcgatact gttcaactac acgccggcca agagaaccct
     2521 ggtgacaatg ctcacagatc cagagctgta ccaatttacg ccaccacttc ttatgttttc
     2581 gaaaactcta agcatggttc gcaattgttt ggtctagaag ttccaggtta cgtctattcc
     2641 cgtttccaaa acccaaccag taatgttttg gaagaaagaa ttgctgcttt agaaggtggt
     2701 gctgctgctt tggctgtttc ctccggtcaa gccgctcaaa cccttgccat ccaaggtttg
     2761 gcacacactg gtgacaacat cgtttccact tcttacttat acggtggtac ttataaccag
     2821 ttcaaaatct cgttcaaaag atttggtatc gaggctagat ttgttgaagg tgacaatcca
     2881 gaagaattcg aaaaggtctt tgatgaaaga accaaggctg tttatttgga aaccattggt
     2941 aatccaaagt acaatgttcc ggattttgaa aaaattgttg caattgctca caaacacggt
     3001 attccagttg tcgttgacaa cacatttggt gccggtggtt acttctgtca gccaattaaa
     3061 tacggtgctg atattgtaac acattctgct accaaatgga ttggtggtca tggtactact
     3121 atcggtggta ttattgttga ctctggtaag ttcccatgga aggactaccc agaaaagttc
     3181 cctcaattct ctcaacctgc cgaaggatat cacggtacta tctacaatga agcctacggt
     3241 aacttggcat acatcgttca tgttagaact gaactattaa gagatttggg tccattgatg
     3301 aacccatttg cctctttctt gctactacaa ggtgttgaaa cattatcttt gagagctgaa
     3361 agacacggtg aaaatgcatt gaagttagcc aaatggttag aacaatcccc atacgtatct
     3421 tgggtttcat accctggttt agcatctcat tctcatcatg aaaatgctaa gaagtatcta
     3481 tctaacggtt tcggtggtgt cttatctttc ggtgtaaaag acttaccaaa tgccgacaag
     3541 gaaactgacc cattcaaact ttctggtgct caagttgttg acaatttaaa gcttgcctct
     3601 aacttggcca atgttggtga tgccaagacc ttagtcattg ctccatactt cactacccac
     3661 aaacaattaa atgacaaaga aaagttggca tctggtgtta ccaaggactt aattcgtgtc
     3721 tctgttggta tcgaatttat tgatgacatt attgcagact tccagcaatc ttttgaaact
     3781 gttttcgctg gccaaaaacc atgagtgtgc gtaatgagtt gtaaaattat gtataaacct
     3841 actttctctc acaagtacta tacttttata aaacgaactt tattgaaatg aatatccttt
     3901 ttttcccttg ttacatgtcg tgactcgtac tttgaaccta aattgttcta acatcaaaga
     3961 acagtgttaa ttcgcagtcg agaagaaaaa tatggtgaac aagactcatc tacttcatga
     4021 gactacttta cgcctcctat aaagctgtca cactggataa atttattgta ggaccaagtt
     4081 acaaaagagg atgatggagg tttctttaca ataaagaagc acatgtgtgt taacgttttt
     4141 agtatttgct tgttatgtaa atcaggaaaa cttcgcggga tttggttgga tgctactttc
     4201 catacaataa atattataga tctaaaaagc caaattacaa gtaaagatta gtaaagctgt
     4261 tggaattcca tcgttgataa aaatgttagt tattaaatat aaaagtcaga ataggtgaac
     4321 ttggatttaa ttgttggcat ttcgttgctg ctagaggcca taatattaga tagccaggac
     4381 atactagttc tcctcgtggt ataggaatcc ataaaatgga attggtgatt ctatgtgata
     4441 tattcacatt cttactacat tatcaatcct tgcacttcag cttcctctaa cctcgatgac
     4501 atcttctcat aacttatgtc atcatctaac gccgtctatt ataatatatt gatagtataa
     4561 gtattagttg atagacaata gtggattttt attccaacag tgtctttgtt cgtctcagat
     4621 atagtcggat tgccctttta agcaatcaat agtgttttat ttgcaacaat gtcgtcatag
     4681 tttaatatgt cctataagat gttaacttgc tcaacattca acaaagtttg gtctcttggc
     4741 ccagttggtt aaggcaccgt gctaataacg cggggatcag cggttcgatc ccgctagaga
     4801 ccatttattt ttaaattcga ccgtctgaca gacggtgcac ttattactta attttttttt
     4861 ttttttaatc ctcgttaaac tgagtacata atcagcaaca atataatata aaagttttgc
     4921 ttttttatgt acgtacaata ggggtactta aataaataaa taaacaataa aacaatcaat
     4981 aaataaataa tatacgatca taaattacga gatatatttc atagtcttga cttctccagt
     5041 cgacggtatc gataagcttg atatcgaatt cctgcagccc gggggatcca ctagttctag
     5101 agcggccgcc accgcggtgg agctccagct tttgttccct ttagtgaggg ttaattgcgc
     5161 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     5221 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5281 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5341 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5401 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5461 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5521 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5581 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5641 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5701 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5761 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5821 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5881 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5941 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6001 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6061 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6121 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6181 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6241 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6301 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6361 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6421 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6481 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6541 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6601 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6661 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6721 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6781 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6841 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6901 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6961 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7021 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7081 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7141 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7201 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7261 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7321 tgccac
//
