LOCUS       sequence-267532-        6963 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of mTagBFP2-SKL as an additional copy in genome, use 
            auxotrophic marker TRP1(Saccharomyces cerevisiae). Marker for yeast 
            peroxisome..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6963)
  AUTHORS   Zhu J, Zhang ZT, Tang SW, Zhao BS, Li H, Song JZ, Li D, Xie Z
  TITLE     A Validated Set of Fluorescent-Protein-Based Markers for Major 
            Organelles in Yeast (Saccharomyces cerevisiae).
  JOURNAL   MBio. 2019 Sep 3;10(5). pii: mBio.01691-19. doi: 
            10.1128/mBio.01691-19.
  PUBMED    31481383
REFERENCE   2  (bases 1 to 6963)
  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: "MBio."; 
            date: "2019-09-3"; volume: "10(5). pii: mBio.01691-19. doi"; pages: 
            " 10.1128/mBio.01691-19"
COMMENT     Sequence Label: ZJOM160
FEATURES             Location/Qualifiers
     source          1..6963
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     10..29
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        10..28
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        513..1032
                     /gene="S. cerevisiae TPI1"
                     /label=TPI1 promoter
                     /note="strong constitutive promoter for yeast triose 
                     phosphate isomerase"
     primer_bind     complement(1805..1821)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1805..1821)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2803..2823)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2803..2821)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2842..2858)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2842..2858)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2855..2877)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2866..2882
                     /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(2890..2896,2897..2914,2915..2920))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2935..2956
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3073..3090)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3244..3832)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3324..3343)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4003..4794,4795..4863))
                     /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     4626..4645
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4864..4968)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5036..5054
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5092..5114)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5214..5233
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5264..5545
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             5546..6220
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(6351..6806)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6488..6509)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6700..6719
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6932..6954
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6946..6963
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6947..6963
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gagcgcgcgt aatacgactc actatagggc gaattgggta cccaagaata agggaacaag
       61 attttggtaa tttccaaaaa atcaatagca tgcaggacgt tatgaagaag agatctacgt
      121 atggtcattt cttcttcaga ttccctcatg gagaaagtgc ggcagatgta tatgacagag
      181 tcgccagttt ccaagagact ttattcaggc acttccatga taggcaagag agaagaccca
      241 gagatgttgt tgtcctagtt acacatggta tttattccag agtattcctg atgaaatggt
      301 ttagatggac atacgaagag tttgaatcgt ttaccaatgt tcctaacggg agcgtaatgg
      361 tgatggaact ggacgaatcc atcaatagat acgtcctgag gaccgtgcta cccaaatgga
      421 ctgattgtga gggagaccta actacatagt gtttaaagat tacggatatt taacttactt
      481 agaataatgc catttttttg agttataata atcctacgtt agtgtgagcg ggatttaaac
      541 tgtgaggacc ttaatacatt cagacacttc tgcggtatca ccctacttat tcccttcgag
      601 attatatcta ggaacccatc aggttggtgg aagattaccc gttctaagac ttttcagctt
      661 cctctattga tgttacacct ggacacccct tttctggcat ccagttttta atcttcagtg
      721 gcatgtgaga ttctccgaaa ttaattaaag caatcacaca attctctcgg ataccacctc
      781 ggttgaaact gacaggtggt ttgttacgca tgctaatgca aaggagccta tatacctttg
      841 gctcggctgc tgtaacaggg aatataaagg gcagcataat ttaggagttt agtgaacttg
      901 caacatttac tattttccct tcttacgtaa atatttttct ttttaattct aaatcaatct
      961 ttttcaattt tttgtttgta ttcttttctt gcttaaatct ataactacaa aaaacacata
     1021 cataaactaa aaatggctag aactttcttt gtcgacatga gcgaactgat caaagagaac
     1081 atgcacatga agctgtacat ggaaggcacc gttgacaacc accactttaa gtgcacctct
     1141 gagggtgagg gtaagcccta cgaaggcacc caaaccatgc gtatcaaagt tgtggagggc
     1201 ggtccactgc cattcgcttt tgacattctg gccaccagct tcctgtacgg ttccaaaacc
     1261 ttcattaacc atactcaggg cattcctgat ttcttcaaac agagctttcc ggaaggtttc
     1321 acctgggagc gtgtcaccac gtatgaagat ggtggtgtgt tgaccgccac ccaagatacc
     1381 tccctgcaag atggctgtct gatctataac gtgaaaattc gtggcgtcaa ctttactagc
     1441 aatggtcccg tgatgcagaa gaaaaccctg ggttgggagg cctttacaga aaccctgtat
     1501 cctgccgatg gtggcctgga gggccgtaac gacatggcac tgaagctggt tggtggcagc
     1561 catttgatcg caaatgccaa gaccacttac agaagcaaga aacccgcaaa aaatctgaag
     1621 atgccgggtg tttactatgt cgactaccgt ctggaaagaa ttaaagaagc aaataatgag
     1681 acttacgtgg agcagcacga ggttgcagtc gcgagatatt gcgacttgcc tagcaagctg
     1741 ggtcataaac tgaattccaa actatagatc gataagcttg atatcgaatt cctgcagccc
     1801 gggggatcca ctagttctag aaactaagat taatataatt atataaaaat attatcttct
     1861 tttctttata tctagtgtta tgtaaaataa attgatgact acggaaagct tttttatatt
     1921 gtttcttttt cattctgagc cacttaaatt tcgtgaatgt tcttgtaagg gacggtagat
     1981 ttacaagtga tacaacaaaa agcaaggcgc tttttctaat aaaaagaaga aaagcattta
     2041 acaattgaac acctctatat caacgaagaa tattactttg tctctaaatc cttgtaaaat
     2101 gtgtacgatc tctatatggg ttactcataa gtgtaccgaa gactgcattg aaagtttatg
     2161 ttttttcact ggaggcgtca ttttcgcgtt gagaagatgt tcttatccaa atttcaactg
     2221 ttatatagaa gagcaaaaaa ttgccaaaaa aaacaacatt tattcatttg aaatataaaa
     2281 tttgggcttc tatattttaa tattgctttt caattactgt tattaaatgt aagtactgtg
     2341 tctatgaaaa tatatgcaaa tgctaagaaa aatcctaaaa atttgaatat gagatattcc
     2401 tcagtatttc tttttcatcc tttcttctgc ggctctagcc ctttgttctc tcatcaatct
     2461 gcgtctctgt tcatcggtca aagaattcaa attttgttgc tgaattgaag gaatattggt
     2521 taactttttg cctggacccc tctctgcatc tttagcaaat ctttgttttt ccttggcggc
     2581 ttctaattcc aatttaatca gtttttctgc cttttcattc tctattgctt ttttcttttc
     2641 attgatttca tcaacttgtt ctctgctcaa tggctgaccg atatgagttt cgtcccaatt
     2701 agtgaaagac tctccaagat tgtatcttgc aatttgaaac gcgtgtctca cgtcgtcgtt
     2761 tctattcaaa tcccacccgc ggtggagctc cagcttttgt tccctttagt gagggttaat
     2821 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     2881 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     2941 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     3001 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3061 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3121 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3181 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3241 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3301 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3361 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3421 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3481 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3541 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3601 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3661 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3721 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3781 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3841 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3901 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3961 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4021 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4081 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4141 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4201 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4261 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4321 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4381 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4441 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4501 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4561 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4621 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4681 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4741 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4801 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4861 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4921 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4981 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     5041 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     5101 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     5161 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     5221 agagcagatt gtactgagag tgcaccatac cacagctttt caaaacgaca ttactatata
     5281 tataatatag gaagcattta atagaacagc atcgtaatat atgtgtactt tgcagttatg
     5341 acgccagatg gcagtagtgg aagatattct ttattgaaaa atagcttgtc accttacgta
     5401 caatcttgat ccggagcttt tctttttttg ccgattaaga attaattcgg tcgaaaaaag
     5461 aaaaggagag ggccaagagg gagggcattg gtgactattg agcacgtgag tatacgtgat
     5521 taagcacaca aaggcagctt ggagtatgtc tgttattaat ttcacaggta gttctggtcc
     5581 attggtgaaa gtttgcggct tgcagagcac agaggccgca gaatgtgctc tagattccga
     5641 tgctgacttg ctgggtatta tatgtgtgcc caatagaaag agaacaattg acccggttat
     5701 tgcaaggaaa atttcaagtc ttgtaaaagc atataaaaat agttcaggca ctccgaaata
     5761 cttggttggc gtgtttcgta atcaacctaa ggaggatgtt ttggctctgg tcaatgatta
     5821 cggcattgat atcgtccaac tgcatggaga tgagtcgtgg caagaatacc aagagttcct
     5881 cggtttgcca gttattaaaa gactcgtatt tccaaaagac tgcaacatac tactcagtgc
     5941 agcttcacag aaacctcatt cgtttattcc cttgtttgat tcagaagcag gtgggacagg
     6001 tgaacttttg gattggaact cgatttctga ctgggttgga aggcaagaga gccccgaaag
     6061 cttacatttt atgttagctg gtggactgac gccagaaaat gttggtgatg cgcttagatt
     6121 aaatggcgtt attggtgttg atgtaagcgg aggtgtggag acaaatggtg taaaagactc
     6181 taacaaaata gcaaatttcg tcaaaaatgc taagaaatag aaaactgtat tataagtaaa
     6241 tgcatgtata ctaaactcac aaattagagc ttcaatttaa ttatatcagt tattacccta
     6301 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag
     6361 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     6421 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     6481 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     6541 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     6601 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     6661 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     6721 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     6781 gcttaatgcg ccgctacagg gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa
     6841 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     6901 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     6961 agt
//
