LOCUS       sequence-254023-        4963 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast pathway position 1. ITE transcription unit with the PRE3 
            promoter and ENO1 terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4963)
  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 4963)
  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-5a-Ppre3-ITE-Teno1
FEATURES             Location/Qualifiers
     source          1..4963
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    136..169
                     /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)"
     protein_bind    complement(2406..2439)
                     /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(2909..2926)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2909..2925)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3024..3041)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3195..3783)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3275..3294)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3905..4714)
                     /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(4715..4806)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4873..4892
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ccgatggtag tgtggggact ccccatgcga gagtagggaa ctgccaggca tcaaataaaa
       61 cgaaaggctc agtcgaaaga ctgggccttt cgcccgggct aattaggggg tgtcgccctt
      121 attcgactct atagtgaagt tcctattctc tagaaagtat aggaacttct gaagtgggga
      181 agcgacttcc aatcgctttg catatccagt accacaccca caggcgtttg tgccaaacat
      241 taatttgttc tgcatacttt gaacctttca gaaaataaaa aacattacgc gcatacttac
      301 cctgctcgcg aagaagagta acactaacgc attctatggg caattgatga cagtattcag
      361 tacaagacat agtccgtttc cttgattcaa ttcctatagc attatgaact agccgccttt
      421 aagagtgcca agctgttcaa caccgatcat ttttgatgat ttggcgtttt tgttatattg
      481 atagatttct tttgaatttt gtcattttca cttttccact cgcaacggaa tccggtggca
      541 aaaaagggaa aagcattgaa atgcaatctt taacagtatt ttaaacaagt tgcgagacgg
      601 tgtacaatta cgataagaat tgctacttca aagtacacac agaaagttaa catgaatgga
      661 attcaagtgg acatcaatcg tttgaaaaag ggcgaagtca gtttaggtac ctcaatgtat
      721 gtatataaga atttttcctc ccactttatt gtttctaaaa gttcaatgaa gtaaagtctc
      781 aattggcctt attactaact aataggtatc ttataatcac ctaataaaat agaatgggtc
      841 acggtgacac tgaatctcca aacccaacca ccaccactga aggttctggt caaaacgaac
      901 ctgaaaagaa gggtcgtgac attccattat ggagaaagtg tgttatcact ttcgttgttt
      961 cctggatgac tttggttgtc actttctcct ccacctgttt gttgccagct gctccagaaa
     1021 ttgctaacga attcgatatg accgtcgaaa ccattaacat ttccaacgct ggtgttttgg
     1081 ttgccatggg ttactcttct ttgatctggg gtccaatgaa caaattggtt ggtagaagaa
     1141 cctcttacaa cttggccatc tccatgttgt gtgcctgttc tgctggtact gctgctgcca
     1201 tcaacgaaga aatgttcatt gctttccgtg tcttgtctgg cttgaccggt acttctttca
     1261 tggtttccgg tcaaaccgtc ttggctgata tctttgaacc agtttacaga ggtactgctg
     1321 tcggtttctt catggctggt actctatccg gtccagccat tggtccatgt gtcggtggtg
     1381 tcattgtcac tttcacctcc tggagagtta tcttctggtt acaattgggt atgtctggtt
     1441 taggtttggt tttgtctcta ttattcttcc caaagatcga aggtaactct gaaaaggttt
     1501 ctactgcttt caagccaacc actttggtca ccatcatctc caagttctct ccaaccgatg
     1561 tcttgaagca atgggtttac ccaaatgtct ttttggctga tttgtgttgt ggtttgttgg
     1621 ccatcactca atactccatc ttgacttctg ccagagctat cttcaactcc agattccatt
     1681 tgaccaccgc tttggtttcc ggtttattct acttggctcc aggtgctggt ttcttgattg
     1741 gttctttggt tggtggtaaa ttgtctgaca gaaccgtcag aagatacatt gtcaagagag
     1801 gtttcagatt acctcaagac agattgcact ctggtttgat cactttgttt gctgtcttgc
     1861 cagctggtac tttgatctac ggttggactt tgcaagagga caagggtgac atggttgttc
     1921 caatcattgc tgctttcttt gctggttggg gtttgatggg ttctttcaac tgtttgaaca
     1981 cctacgttgc tggtttattc cacactttga tctacttgtt cccattgtgt acctgtccac
     2041 aataaaagct tttgattaag ccttctagtc caaaaaacac gtttttttgt catttatttc
     2101 attttcttag aatagtttag tttattcatt ttatagtcac gaatgtttta tgattctata
     2161 tagggttgca aacaagcatt tttcatttta tgttaaaaca atttcaggtt taccttttat
     2221 tctgcttgtg gtgacgcgtg tatccgcccg ctcttttggt cacccatgta tttaattgca
     2281 taaataattc ttaaaagtgg agctagtcta tttctattta catacctctc atttctcatt
     2341 tcctcttgcc catcgaacgt acaagtactc ctctgttctc tccttccttt gctttaaaat
     2401 gaagtgaagt tcctatactt tctagagaat aggaacttct atagtgagtc gaataagggc
     2461 gacacaaaat ttattctaaa tgcataataa atactgataa catcttatag tttgtattat
     2521 attttgtatt atcgttgaca tgtataattt tgatatcaaa aactgatttt ccctttatta
     2581 ttttcgagat ttattttctt aattctcttt aacaaactag aaatattgta tatacaaaaa
     2641 atcataaata atagatgaat agtttaatta taggtgttca tcaatcgaaa aagcaacgta
     2701 tcttatttaa agtgcgttgc ttttttctca tttataaggt taaataattc tcatatatca
     2761 agcaaagtga caggcgccct taaatattct gacaaatgct ctttccctaa actcccccca
     2821 taaaaaaacc cgccgaagcg ggtttttacg ttatttgcgg attaacgatt actcgttatc
     2881 agaaccgccc agggggcccg agcttaagac tggccgtcgt tttacaacac agaaagagtt
     2941 tgtagaaacg caaaaaggcc atccgtcagg ggccttctgc ttagtttgat gcctggcagt
     3001 tccctactct cgccttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3061 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3121 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3181 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3241 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3301 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3361 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3421 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3481 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3541 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3601 ttgaagtggt gggctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3661 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3721 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3781 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga cgcgcgcgta
     3841 actcacgtta agggattttg gtcatgagct tgcgccgtcc cgtcaagtca gcgtaatgct
     3901 ctgcttagaa aaactcatcg agcatcaaat gaaattgcaa tttattcata tcaggattat
     3961 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     4021 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     4081 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     4141 cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag
     4201 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     4261 attgcgcctg agcgaggcga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     4321 tcgagtgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     4381 gatattcttc taatacctgg aacgctgttt ttccggggat cgcagtggtg agtaaccatg
     4441 catcatcagg agtacggata aaatgcttga tggtcggaag tggcataaat tccgtcagcc
     4501 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     4561 gaaacaactc tggcgcatcg ggcttcccat acaagcgata gattgtcgca cctgattgcc
     4621 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     4681 gcggcctcga cgtttcccgt tgaatatggc tcatattctt cctttttcaa tattattgaa
     4741 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4801 aacaaatagg ggtcagtgtt acaaccaatt aaccaattct gaacattatc gcgagcccat
     4861 ttatacctga atatggctca taacacccct tgtttgcctg gcggcagtag cgcggtggtc
     4921 ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcg
//
