LOCUS       sequence-137235-        4377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  standard plasmids for exogenus pathway assembly haboring yeast 
            auxotrophic marker LEU2 which could be released and ligated to 
            series of TUs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4377)
  AUTHORS   Guo Y, Dong J, Zhou T, Auxillos J, Li T, Zhang W, Wang L, Shen Y, 
            Luo Y, Zheng Y, Lin J, Chen GQ, Wu Q, Cai Y, Dai J
  TITLE     YeastFab: the design and construction of standard biological parts 
            for metabolic engineering in Saccharomyces cerevisiae.
  JOURNAL   Nucleic Acids Res. 2015 May 8. pii: gkv464.
  PUBMED    25956650
REFERENCE   2  (bases 1 to 4377)
  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: "Nucleic 
            Acids Res. 2015 May 8. pii: gkv464."
COMMENT     Sequence Label: pMV-LEU2
FEATURES             Location/Qualifiers
     source          1..4377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     94..112
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     121..138
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     122..138
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        444..851
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             852..1946
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(2480..2496)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2480..2496)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2508..2525)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2679..3267)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2759..2778)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3438..4229,4230..4298))
                     /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     4061..4080
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4299..4377,1..26))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
       61 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
      121 tgtaaaacga cggccagtcg tctcgatccg cagtgtcttg cgtctcttga gctgtgcggt
      181 atttcacacc gcatatcgac cggtcgagga gaacttctag tatatctaca tacctaatat
      241 tattgcctta ttaaaaatgg aatcccaaca attacatcaa aatccacatt ctcttcaaaa
      301 tcaattgtcc tgtacttcct tgttcatgtg tgttcaaaaa cgttatattt ataggataat
      361 tatactctat ttctcaacaa gtaattggtt gtttggccga gcggtctaag gcgcctgatt
      421 caagaaatat cttgaccgca gttaactgtg ggaatactca ggtatcgtaa gatgcaagag
      481 ttcgaatctc ttagcaacca ttattttttt cctcaacata acgagaacac acaggggcgc
      541 tatcgcacag aatcaaattc gatgactgga aattttttgt taatttcaga ggtcgcctga
      601 cgcatatacc tttttcaact gaaaaattgg gagaaaaagg aaaggtgaga gcgccggaac
      661 cggcttttca tatagaatag agaagcgttc atgactaaat gcttgcatca caatacttga
      721 agttgacaat attatttaag gacctattgt tttttccaat aggtggttag caatcgtctt
      781 actttctaac ttttcttacc ttttacattt cagcaatata tatatatata tttcaaggat
      841 ataccattct aatgtctgcc cctaagaaga tcgtcgtttt gccaggtgac cacgttggtc
      901 aagaaatcac agccgaagcc attaaggttc ttaaagctat ttctgatgtt cgttccaatg
      961 tcaagttcga tttcgaaaat catttaattg gtggtgctgc tatcgatgct acaggtgtcc
     1021 cacttccaga tgaggcgctg gaagcctcca agaaggttga tgccgttttg ttaggtgctg
     1081 tgggtggtcc taaatggggt accggtagtg ttagacctga acaaggttta ctaaaaatcc
     1141 gtaaagaact tcaattgtac gccaacttaa gaccatgtaa ctttgcatcc gactctcttt
     1201 tagacttatc tccaatcaag ccacaatttg ctaaaggtac tgacttcgtt gttgtcagag
     1261 aattagtggg aggtatttac tttggtaaga gaaaggaaga cgatggtgat ggtgtcgctt
     1321 gggatagtga acaatacacc gttccagaag tgcaaagaat cacaagaatg gccgctttca
     1381 tggccctaca acatgagcca ccattgccta tttggtcctt ggataaagct aatgttttgg
     1441 cctcttcaag attatggaga aaaactgtgg aggaaaccat caagaacgaa ttccctacat
     1501 tgaaggttca acatcaattg attgattctg ccgccatgat cctagttaag aacccaaccc
     1561 acctaaatgg tattataatc accagcaaca tgtttggtga tatcatctcc gatgaagcct
     1621 ccgttatccc aggttccttg ggtttgttgc catctgcgtc cttggcctct ttgccagaca
     1681 agaacaccgc atttggtttg tacgaaccat gccacggttc tgctccagat ttgccaaaga
     1741 ataaggtcaa ccctatcgcc actatcttgt ctgctgcaat gatgttgaaa ttgtcattga
     1801 acttgcctga agaaggtaag gccattgaag atgcagttaa aaaggttttg gatgcaggta
     1861 tcagaactgg tgatttaggt ggttccaaca gtaccaccga agtcggtgat gctgtcgccg
     1921 aagaagttaa gaaaatcctt gcttaaaaag attctctttt tttatgatat ttgtacataa
     1981 actttataaa tgaaattcat aatagaaacg acacgaaatt acaaaatgga atatgttcat
     2041 agggtagacg aaactatata cgcaatctac atacatttat caagaaggag aaaaaggagg
     2101 atgtaaagga atacaggtaa gcaaattgat actaatggct caacgtgata aggaaaaaga
     2161 attgcacttt aacattaata ttgacaagga ggagggcacc acacaaaaag ttaggtgtaa
     2221 cagaaaatca tgaaactatg attcctaatt tatatattgg aggattttct ctaaaaaaaa
     2281 aaaaatacaa caaataaaaa acactcaatg acctgaccat ttgatggagt ttaagtcaat
     2341 accttcttga accatttccc ataatggtga aagttccctc aagaatttta ctctgtcaga
     2401 aacggcctta acgacgtagt cgatatggtg cactctcagt acaatctcgc tagagacgga
     2461 gtcactgcca accgagacgg tcatagctgt ttcctgtgtg ccgcttcctc gctcactgac
     2521 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2581 cggttaccca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2641 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2701 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2761 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2821 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     2881 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2941 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3001 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3061 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     3121 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3181 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3241 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3301 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3361 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3421 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3481 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3541 ggcttaccat ctggccccag tgctgcaata ataccgcggg acccacgctc accggctcca
     3601 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3661 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3721 gttaatagtt tgcgcaacgt tgttgccatc gctacaggca tcgtggtgtc acgctcgtcg
     3781 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     3841 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     3901 gccgccgtgt tatcactcat ggttatggca gcactacata attctcttac tgtcatgcca
     3961 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4021 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     4081 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4141 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4201 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4261 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4321 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttag
//
