LOCUS       sequence-254154-        4093 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast pathway position 2. NatMX transcription unit with the S. 
            paradoxus TEF1 promoter and TIP1 terminator. Nourseothricin 
            resistance..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4093)
  AUTHORS   Young EM, Zhao Z, Gielesen BEM, Wu L, Benjamin Gordon D, Roubos JA, 
            Voigt CA
  TITLE     Iterative algorithm-guided design of massive strain libraries, 
            applied to itaconic acid production in yeast.
  JOURNAL   Metab Eng. 2018 Jul;48:33-43. doi: 10.1016/j.ymben.2018.05.002. Epub
            2018 May 9.
  PUBMED    29753070
REFERENCE   2  (bases 1 to 4093)
  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: "Metab 
            Eng."; date: "2018-07"; volume: "48:33-43. doi"; pages: " 
            10.1016/j.ymben.2018.05.002. Epub 2018 May 9"
COMMENT     Sequence Label: pDSM-ab-Psptef1-NAT-Ttip1
FEATURES             Location/Qualifiers
     source          1..4093
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             393..965
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     protein_bind    complement(1280..1313)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(1783..1800)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1783..1799)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1898..1915)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2069..2657)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2149..2168)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2779..3588)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        complement(3589..3680)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3747..3766
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    3973..4006
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
ORIGIN
        1 tttttttact cttccgcatt ttctcggact gcgcgcatcg ccgcaccgct tccaaaacac
       61 ctgaacatta catactattt ttcccctctt tctttcttta gggtggtgtt aaattacccg
      121 ctctgaagct ttggaaaaga aacaaaaggc cacttcgttt ctttttcttc gtcgaaaagg
      181 gcaaaaaaaa aatttttacc acgtttcttt ttcttgaaaa tttttttttt ttattttttc
      241 tctttcgatg acctcccatt gatatttaag ttaataaatg gtcatcaatt tctcaagttt
      301 tattttcgtt ttccttgttt catgtcgact tttttacatc cttctcagtt agaaagaaag
      361 catagcaatc taatctaagt tttaattaca aaatgggtac cactcttgac gacacggctt
      421 accggtaccg caccagtgtc ccgggggacg ccgaggccat cgaggcactg gatgggtcct
      481 tcaccaccga caccgtattc cgcgtcaccg ccaccgggga cggcttcacc ctgcgggagg
      541 tgccggtgga cccgcccctg accaaggtgt tccccgacga cgaatcggac gacgaatcgg
      601 acgacgggga ggacggcgac ccggattccc ggacgttcgt cgcgtacggg gacgacggcg
      661 acctggcggg cttcgtggtc gtctcgtact ccggctggaa ccgccggctg accgtcgagg
      721 acatcgaggt cgccccggag caccgggggc acggggtcgg gcgcgcgttg atggggctcg
      781 cgacggagtt cgcccgcgag cggggcgccg ggcacctctg gctggaggtc accaacgtca
      841 acgcaccggc gatccacgcg taccggcgga tggggttcac cctctgcggc ctggacaccg
      901 ccctgtacga cggcaccgcc tcggacggcg agcaggcgct ctacatgagc atgccctgcc
      961 cctaaaaggg aaccttttac aacaaatatt tgaaaaatta cctccattat tataccttct
     1021 ctttatgtaa ttgttagttc gaaaattttt tcttcattaa tataatcaac ttctaaaact
     1081 ttctaaaaac gttctctttt tcgagattag tgcttcttcc caatccgtaa gaaatgtttc
     1141 ctttcttgac aattggcacc agctggctac tcgttgctcg aaaactactc tcttttattt
     1201 ttaatttacg aacgacctcc ggatcgatgt acacaaccga ctgcacccaa acgaacacaa
     1261 atcttagcaa aaatgaagtg aagttcctat actttctaga gaataggaac ttctatagtg
     1321 agtcgaataa gggcgacaca aaatttattc taaatgcata ataaatactg ataacatctt
     1381 atagtttgta ttatattttg tattatcgtt gacatgtata attttgatat caaaaactga
     1441 ttttcccttt attattttcg agatttattt tcttaattct ctttaacaaa ctagaaatat
     1501 tgtatataca aaaaatcata aataatagat gaatagttta attataggtg ttcatcaatc
     1561 gaaaaagcaa cgtatcttat ttaaagtgcg ttgctttttt ctcatttata aggttaaata
     1621 attctcatat atcaagcaaa gtgacaggcg cccttaaata ttctgacaaa tgctctttcc
     1681 ctaaactccc cccataaaaa aacccgccga agcgggtttt tacgttattt gcggattaac
     1741 gattactcgt tatcagaacc gcccaggggg cccgagctta agactggccg tcgttttaca
     1801 acacagaaag agtttgtaga aacgcaaaaa ggccatccgt caggggcctt ctgcttagtt
     1861 tgatgcctgg cagtttccta ctctcgcctt ccgcttcctc gctcactgac tcgctgcgct
     1921 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     1981 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2041 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2101 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2161 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2221 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2281 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2341 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2401 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2461 gtgctacaga gttcttgaag tggtgggcta actacggcta cactagaaga acagtatttg
     2521 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2581 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2641 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2701 acgacgcgcg cgtaactcac gttaagggat tttggtcatg agcttgcgcc gtcccgtcaa
     2761 gtcagcgtaa tgctctgctt agaaaaactc atcgagcatc aaatgaaatt gcaatttatt
     2821 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     2881 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     2941 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     3001 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
     3061 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     3121 gttattcatt cgtgattgcg cctgagcgag gcgaaatacg cgatcgctgt taaaaggaca
     3181 attacaaaca ggaatcgagt gcaaccggcg caggaacact gccagcgcat caacaatatt
     3241 ttcacctgaa tcaggatatt cttctaatac ctggaacgct gtttttccgg ggatcgcagt
     3301 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagtggcat
     3361 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     3421 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaagc gatagattgt
     3481 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     3541 gttggaattt aatcgcggcc tcgacgtttc ccgttgaata tggctcatat tcttcctttt
     3601 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     3661 tatttagaaa aataaacaaa taggggtcag tgttacaacc aattaaccaa ttctgaacat
     3721 tatcgcgagc ccatttatac ctgaatatgg ctcataacac cccttgtttg cctggcggca
     3781 gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg
     3841 atggtagtgt ggggactccc catgcgagag tagggaactg ccaggcatca aataaaacga
     3901 aaggctcagt cgaaagactg ggcctttcgc ccgggctaat tagggggtgt cgcccttatt
     3961 cgactctata gtgaagttcc tattctctag aaagtatagg aacttctgaa gtggggttgc
     4021 ccatcgaacg tacaagtact cctctgttct ctccttcctt tgctttgtgc atagccgaca
     4081 aatcttttac tcc
//
