LOCUS       sequence-267533-        6714 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of MNN9-mTagBFP2 as an additional copy in genome, use 
            auxotrophic marker TRP1(Kluyveromyces lactis). Marker for yeast 
            early golgi..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6714)
  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 6714)
  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: ZJOM100
FEATURES             Location/Qualifiers
     source          1..6714
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      2554..2741
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2609..2633)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    2793..2826
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3829..3862
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(3915..3934)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3916..3934)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4021..4038)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4192..4780)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4272..4291)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4951..5742,5743..5811))
                     /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     5574..5593
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5812..5916)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5984..6002
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6040..6062)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6162..6181
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6262..6280
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6262..6279
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gagctctagc aatgtagaca agaaaatgtt tatgagcata tgtaataaag tatggaatga
       61 atcaaaggag ctgacaaaaa aatcatcatc acatcacaga accggaaaat agtagccaca
      121 caggcactta taaacttaca tacactcgca tacactcacc tactcacaca cgcactcaca
      181 cacacactct cactcgcaca ccaaggacaa ttcagtacat acacaatttt tatataagtt
      241 tgataaacta aaagatgagg gtgagtattc accttattaa tagttacagc ctttatcgtc
      301 actagacacg acagcattgg tttgtctttt tttttttttt ttttcctttc catcgttgta
      361 gcggatttgg cgttgaagta ttttgatata agtaaaacag aacaccaaat caagaataat
      421 aaaggaggaa aagtcgtcaa ccaagcggat tctcaattgg tagaaaagct tttattattt
      481 tctgttagcc gctcttggga cccgttattt ctgaacctat ttttccttcc gcactttgtg
      541 aaagtaagaa gagaatagca tcaattttag tatttttcaa acagaagaat ttaaaagagc
      601 tagaataaaa agttaggaaa caatgtcact ttctcttgta tcgtaccgcc taagaaagaa
      661 cccgtgggtt aacatttttc tacctgtttt ggccatattt ctaatatata taattttttt
      721 ccagagagat caatctctgt tgggacttaa tggccagtcc atttcccaac acaaatgggc
      781 acacgaaaag gaaaacacat tttattttcc cttcaccaag aaatacaaaa tgccaaagta
      841 ctcttataag aaaaaaagcg gctggttgtt caacgatcac gtggaagata ttatcccaga
      901 aggtcatatt gcacattatg atttgaacaa attgcactct acgtcagaag cagctgtcaa
      961 taaggagcat attttgatat tgactccaat gcaaacattt catcaacaat actgggacaa
     1021 tttgctgcag ctaaattacc ctcgggaatt gattgaattg gggttcatta caccaagaac
     1081 agccactggt gacttggcct taaagaaatt ggagaatgct attaaaaagg ttcaaacgga
     1141 caagaaaact caaagattta gtaaaattac tattttgcga cagaattccc agagttttga
     1201 taagttgatg gagaaggaaa gacacgcttt agatgttcaa aaggaaagac gtgcagcaat
     1261 ggctttggcg cgcaatgaat tactattctc caccatagga cctcacactt cttgggtgct
     1321 gtggctagat gccgatatta tagagacacc accatcttta attcaagaca tgaccaaaca
     1381 caacaaagct atcttagctg caaacattta tcaaagattt tacgatgaag agaagaagca
     1441 accatcaatc agaccatacg atttcaacaa ctggcaagaa agtgacaccg gtttagaaat
     1501 agcctctcag atgggtgatg acgagattat tgtcgagggt tatgcagaaa ttgccactta
     1561 taggccatta atggctcatt tctacgatgc taatggcgta ccaggtgaag agatggcgct
     1621 ggatggtgtt ggtggaggct gtactttggt caaagcagaa gttcacagag acggtgccat
     1681 gttccctaat ttcccatttt atcacttgat tgaaacagaa ggttttgcta agatggcgaa
     1741 gagattaaac tacgatgtat ttggcttacc aaactatttg gtttatcaca tagaggaaga
     1801 gaaccatctt tctcttgaag ttgctgaaga aatgagcgaa ctgatcaaag agaacatgca
     1861 catgaagctg tacatggaag gcaccgttga caaccaccac tttaagtgca cctctgaggg
     1921 tgagggtaag ccctacgaag gcacccaaac catgcgtatc aaagttgtgg agggcggtcc
     1981 actgccattc gcttttgaca ttctggccac cagcttcctg tacggttcca aaaccttcat
     2041 taaccatact cagggcattc ctgatttctt caaacagagc tttccggaag gtttcacctg
     2101 ggagcgtgtc accacgtatg aagatggtgg tgtgttgacc gccacccaag atacctccct
     2161 gcaagatggc tgtctgatct ataacgtgaa aattcgtggc gtcaacttta ctagcaatgg
     2221 tcccgtgatg cagaagaaaa ccctgggttg ggaggccttt acagaaaccc tgtatcctgc
     2281 cgatggtggc ctggagggcc gtaacgacat ggcactgaag ctggttggtg gcagccattt
     2341 gatcgcaaat gccaagacca cttacagaag caagaaaccc gcaaaaaatc tgaagatgcc
     2401 gggtgtttac tatgtcgact accgtctgga aagaattaaa gaagcaaata atgagactta
     2461 cgtggagcag cacgaggttg cagtcgcgag atattgcgac ttgcctagca agctgggtca
     2521 taaactgaat tgaggcgcgc cacttctaaa taagcgaatt tcttatgatt tatgattttt
     2581 attattaaat aagttataaa aaaaataagt gtatacaaat tttaaagtga ctcttaggtt
     2641 ttaaaacgaa aattcttatt cttgagtaac tctttcctgt aggtcaggtt gctttctcag
     2701 gtatagtatg aggtcgctct tattgaccac acctctaccg gcagatccgc tagggataac
     2761 agggtaatat ctgcaggtcg acaaccctta atataacttc gtataatgta tgctatacga
     2821 agttattagg tctagagcgg ccgcggaacg atcattcact atatatatat caatttatat
     2881 atacgtatgt gtaattgaag aaagatacgt tttttcctct attgagaggc ctgctggatg
     2941 aatagcttta ccttttctaa atccttgata ccatcagtct ctactcctcc acttacatcg
     3001 acaccgatcg catttggtaa catattaatg gcaacagaaa cgttatcagg attcaatcca
     3061 ccagcgataa tgaattttat ctcgggatga cttgcagacc aactggaaat tgcactccaa
     3121 ttcaatttct caccagtgcc accttcacca gaatcgaaca acgtcagcac attgtctacg
     3181 tgttcataca ggtccagtag taattcacaa tcctgtggga actggaacct cttaatgatt
     3241 ggaattgaag atgggatcaa agatctgtat tctttaatat cttcatctcc atgtaattgt
     3301 atcacatcta gattatattc gtggtacagt tgaaggacat catcaacgga ctgatttcta
     3361 aacaccccga ctagtttagt acctttcacg ttctcttgtt ggtgaactgc agttgaaata
     3421 cctttcgcaa cagatgagtc aatggttctt ttcctaccgg gaacacaaat gatacctaag
     3481 taatcagcac catcatccac agcagtcttt gcagcttcaa cggtttgcaa accacacact
     3541 ttaacgagca tcacagtaga tttcgcaaga gggtagctcg ctcagagtac ccaagtaaat
     3601 gattagtaaa ctgatgtttg atagttcaat ttttcaatga aataacctta tattaaaatt
     3661 gatattacta ttatacaaaa ataaagaata aaggatttga gtttatacat aaaataccat
     3721 tattatttgt tcagtgagag atactggggt atatgggatg tgtgtagtga taccatgcaa
     3781 tcatgtatca aacatgggcc cggtaccgag ctctcgagaa cccttaatat aacttcgtat
     3841 aatgtatgct atacgaagtt attaggtgat atcagatcca ctagtggcct atgcggccgc
     3901 ggatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg ttaacctgca
     3961 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4021 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4081 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4141 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4201 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4261 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4321 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4381 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4441 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4501 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4561 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4621 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4681 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4741 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4801 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4861 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4921 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4981 ctcagcgatc tgtctatttc gttcatccat agttgcctga cttcccgtcg tgtagataac
     5041 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5101 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5161 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5221 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5281 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5341 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5401 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5461 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5521 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5581 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5641 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5701 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5761 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5821 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5881 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5941 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     6001 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     6061 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     6121 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     6181 actgagagtg caccatatgg acatattgtc gttagaacgc ggctacaatt aatacataac
     6241 cttatgtatc atacacatac gatttaggtg acactataga acgcggccgc cagctgaagc
     6301 ttcgtacgct gcaggtcgtg ggattcctgg gttaattaac atcgctagcg taggatcaag
     6361 tgcgaatgcc ggtcctacgc aacagcgcgc ggatctacgc ccagcagata tgatacctgc
     6421 tgaagaatac cactttgaac gggatgcatt actctcggcc aacacgaaag ccagaagcgc
     6481 aagcacaagc acaagtacaa gcacaagcac aaaccgggac aggagttcat ggcatgaggc
     6541 agaacccaat aaagacgaag aagagggtac ggacttggcc atcgaggacg gagcggtccc
     6601 cactcccact tttactacgt tccagcggac ctcgcagccg cagcagcaat cacctagtct
     6661 tcttcaaggc gaaatccgac tctcatcgca tatttttgcc ttcgagttcc ccct
//
