LOCUS       sequence-256886-        6966 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 6966)
  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 6966)
  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..6966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        201..481
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             482..1156
                     /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(1287..1742)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1424..1445)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1636..1655
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1872..1894
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1886..1903
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1887..1903
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1913..1932
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1913..1931
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2416..2935
                     /gene="S. cerevisiae TPI1"
                     /label=TPI1 promoter
                     /note="strong constitutive promoter for yeast triose 
                     phosphate isomerase"
     primer_bind     complement(3708..3724)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3708..3724)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4706..4726)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4706..4724)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4745..4761)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4745..4761)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4758..4780)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4769..4785
                     /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(4793..4799,4800..4817,4818..4823))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4976..4993)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5147..5735)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5227..5246)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5906..6697,6698..6766))
                     /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     6529..6548
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6767..6871)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6939..6957
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac agcttttcaa aacgacatta ctatatatat aatataggaa gcatttaata
      241 gacagcatcg taatatatgt gtactttgca gttatgacgc cagatggcag tagtggaaga
      301 tattctttat tgaaaaatag cttgtcacct tacgtacaat cttgatccgg agcttttctt
      361 tttttgccga ttaagaatta attcggtcga aaaaagaaaa ggagagggcc aagagggagg
      421 gcattggtga ctattgagca cgtgagtata cgtgattaag cacacaaagg cagcttggag
      481 tatgtctgtt attaatttca caggtagttc tggtccattg gtgaaagttt gcggcttgca
      541 gagcacagag gccgcagaat gtgctctaga ttccgatgct gacttgctgg gtattatatg
      601 tgtgcccaat agaaagagaa caattgaccc ggttattgca aggaaaattt caagtcttgt
      661 aaaagcatat aaaaatagtt caggcactcc gaaatacttg gttggcgtgt ttcgtaatca
      721 acctaaggag gatgttttgg ctctggtcaa tgattacggc attgatatcg tccaactgca
      781 tggagatgag tcgtggcaag aataccaaga gttcctcggt ttgccagtta ttaaaagact
      841 cgtatttcca aaagactgca acatactact cagtgcagct tcacagaaac ctcattcgtt
      901 tattcccttg tttgattcag aagcaggtgg gacaggtgaa cttttggatt ggaactcgat
      961 ttctgactgg gttggaaggc aagagagccc cgaaagctta cattttatgt tagctggtgg
     1021 actgacgcca gaaaatgttg gtgatgcgct tagattaaat ggcgttattg gtgttgatgt
     1081 aagcggaggt gtggagacaa atggtgtaaa agactctaac aaaatagcaa atttcgtcaa
     1141 aaatgctaag aaatagaaaa ctgtattata agtaaatgca tgtatactaa actcacaaat
     1201 tagagcttca atttaattat atcagttatt accctatgcg gtgtgaaata ccgcacagat
     1261 gcgtaaggag aaaataccgc atcaggaaat tgtaaacgtt aatattttgt taaaattcgc
     1321 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     1381 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     1441 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     1501 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     1561 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     1621 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     1681 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     1741 gtcgcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     1801 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     1861 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga
     1921 ctcactatag ggcgaattgg gtacccaaga ataagggaac aagattttgg taatttccaa
     1981 aaaatcaata gcatgcagga cgttatgaag aagagatcta cgtatggtca tttcttcttc
     2041 agattccctc atggagaaag tgcggcagat gtatatgaca gagtcgccag tttccaagag
     2101 actttattca ggcacttcca tgataggcaa gagagaagac ccagagatgt tgttgtccta
     2161 gttacacatg gtatttattc cagagtattc ctgatgaaat ggtttagatg gacatacgaa
     2221 gagtttgaat cgtttaccaa tgttcctaac gggagcgtaa tggtgatgga actggacgaa
     2281 tccatcaata gatacgtcct gaggaccgtg ctacccaaat ggactgattg tgagggagac
     2341 ctaactacat agtgtttaaa gattacggat atttaactta cttagaataa tgccattttt
     2401 ttgagttata ataatcctac gttagtgtga gcgggattta aactgtgagg accttaatac
     2461 attcagacac ttctgcggta tcaccctact tattcccttc gagattatat ctaggaaccc
     2521 atcaggttgg tggaagatta cccgttctaa gacttttcag cttcctctat tgatgttaca
     2581 cctggacacc ccttttctgg catccagttt ttaatcttca gtggcatgtg agattctccg
     2641 aaattaatta aagcaatcac acaattctct cggataccac ctcggttgaa actgacaggt
     2701 ggtttgttac gcatgctaat gcaaaggagc ctatatacct ttggctcggc tgctgtaaca
     2761 gggaatataa agggcagcat aatttaggag tttagtgaac ttgcaacatt tactattttc
     2821 ccttcttacg taaatatttt tctttttaat tctaaatcaa tctttttcaa ttttttgttt
     2881 gtattctttt cttgcttaaa tctataacta caaaaaacac atacataaac taaaaatggc
     2941 tagaactttc tttgtcgaca tgagcgaact gatcaaagag aacatgcaca tgaagctgta
     3001 catggaaggc accgttgaca accaccactt taagtgcacc tctgagggtg agggtaagcc
     3061 ctacgaaggc acccaaacca tgcgtatcaa agttgtggag ggcggtccac tgccattcgc
     3121 ttttgacatt ctggccacca gcttcctgta cggttccaaa accttcatta accatactca
     3181 gggcattcct gatttcttca aacagagctt tccggaaggt ttcacctggg agcgtgtcac
     3241 cacgtatgaa gatggtggtg tgttgaccgc cacccaagat acctccctgc aagatggctg
     3301 tctgatctat aacgtgaaaa ttcgtggcgt caactttact agcaatggtc ccgtgatgca
     3361 gaagaaaacc ctgggttggg aggcctttac agaaaccctg tatcctgccg atggtggcct
     3421 ggagggccgt aacgacatgg cactgaagct ggttggtggc agccatttga tcgcaaatgc
     3481 caagaccact tacagaagca agaaacccgc aaaaaatctg aagatgccgg gtgtttacta
     3541 tgtcgactac cgtctggaaa gaattaaaga agcaaataat gagacttacg tggagcagca
     3601 cgaggttgca gtcgcgagat attgcgactt gcctagcaag ctgggtcata aactgaattc
     3661 caaactatag atcgataagc ttgatatcga attcctgcag cccgggggat ccactagttc
     3721 tagaaactaa gattaatata attatataaa aatattatct tcttttcttt atatctagtg
     3781 ttatgtaaaa taaattgatg actacggaaa gcttttttat attgtttctt tttcattctg
     3841 agccacttaa atttcgtgaa tgttcttgta agggacggta gatttacaag tgatacaaca
     3901 aaaagcaagg cgctttttct aataaaaaga agaaaagcat ttaacaattg aacacctcta
     3961 tatcaacgaa gaatattact ttgtctctaa atccttgtaa aatgtgtacg atctctatat
     4021 gggttactca taagtgtacc gaagactgca ttgaaagttt atgttttttc actggaggcg
     4081 tcattttcgc gttgagaaga tgttcttatc caaatttcaa ctgttatata gaagagcaaa
     4141 aaattgccaa aaaaaacaac atttattcat ttgaaatata aaatttgggc ttctatattt
     4201 taatattgct tttcaattac tgttattaaa tgtaagtact gtgtctatga aaatatatgc
     4261 aaatgctaag aaaaatccta aaaatttgaa tatgagatat tcctcagtat ttctttttca
     4321 tcctttcttc tgcggctcta gccctttgtt ctctcatcaa tctgcgtctc tgttcatcgg
     4381 tcaaagaatt caaattttgt tgctgaattg aaggaatatt ggttaacttt ttgcctggac
     4441 ccctctctgc atctttagca aatctttgtt tttccttggc ggcttctaat tccaatttaa
     4501 tcagtttttc tgccttttca ttctctattg cttttttctt ttcattgatt tcatcaactt
     4561 gttctctgct caatggctga ccgatatgag tttcgtccca attagtgaaa gactctccaa
     4621 gattgtatct tgcaatttga aacgcgtgtc tcacgtcgtc gtttctattc aaatcccacc
     4681 cgcggtggag ctccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat
     4741 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatag
     4801 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgaggtaa ctcacattaa
     4861 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     4921 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     4981 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5041 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5101 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5161 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5221 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5281 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5341 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5401 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5461 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5521 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5581 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5641 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5701 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5761 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5821 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5881 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5941 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6001 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6061 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6121 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6181 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6241 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6301 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6361 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6421 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6481 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6541 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6601 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6661 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6721 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6781 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6841 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     6901 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     6961 ttcgtc
//
