LOCUS       sequence-253012-        3325 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrative yeast promoter characterization plasmid. Clone in with 
            BpiI. Has ChrXV homology upstream and an mVenus fragment 
            downstream..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3325)
  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 3325)
  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: pEMY01AB-PRO
FEATURES             Location/Qualifiers
     source          1..3325
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    130..151
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(166..171,172..189,190..196)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    204..220
                     /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)."
     primer_bind     209..231
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     228..244
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     228..244
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             240..638
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSILEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISRRIWCRRPAQTP
                     VGVVLDMQSDWKSLRVSKIHTSLHSLTF"
     misc_feature    257..313
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(314..331)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(314..330)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(323..345)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      1041..1070
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     CDS             1233..1892
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1299..1318)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2120..2708
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2609..2628
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttgggacaac accagtgaat aattcttcac ctttagacat taggtcttcc ccgcgcgttg
       61 gccgattcat taatgcagct ggcacgacag gtttccctac tggaaagcgg gcagtgagcg
      121 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
      181 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
      241 tgaccatgat tacgccaagc ttgcatgcct gcaggtcgat tctagaggat ccccgggtac
      301 cgagctcgaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
      361 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
      421 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc
      481 ggtattttct ccttacgcat ctgtgcggta tttcacgccg catatggtgc agaagacctg
      541 cacaaacgcc tgtgggtgtg gtactggata tgcaaagcga ttggaagtcg cttagggttt
      601 caaagatcca tacttctcta cattctctga ctttttaaaa ctgtgtactg gcgaccaatc
      661 gacaaatctg caaccccagc ttcataagct ttctctccca ccagcaaagc ctggacctcc
      721 ctggttctaa taatgagcga ctgaagtttt ccaaaagaaa ccttcactaa acatgttatg
      781 ctcctcaatc acgcgatgaa ataaaggcct attttttgat ccaacgcctt tgaaaatttc
      841 cttatggctg ggtggggtac ctaccatgaa catgttaaag gtaagtggtg gcatattatg
      901 gagttacgcc cgtataccag agtaataatg ttgccgccgg ataggcatgg cgcccccacc
      961 ggattctctt gctatatgga tggcataatt agcttatgaa aggattggta agaatatgtc
     1021 tgagacccaa tcaactcact ggctcacctt cacgggtggg cctttcttcg gcacgggcaa
     1081 attgctgaat attccttttc ttagacgtca ggtggcaaac gcgcgagttg atcgggcacg
     1141 taagaggttc caactttcac cataatgaaa taagatcact accgggcgta ttttttgagt
     1201 tatcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca
     1261 ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc
     1321 aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga
     1381 aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc
     1441 atccggaatt tcgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc
     1501 cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc
     1561 acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa
     1621 acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtctca gccaatccct
     1681 gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg
     1741 ttttcaccat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc
     1801 aggttcatca tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac
     1861 agtactgcga tgagtggcag ggcggggcgt aatttgatat cgagctcgct tggactgctg
     1921 ttgatagatc cagtaatgac ctcagaactc tgtcagacca agtttactca tatatacttt
     1981 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     2041 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     2101 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2161 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2221 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     2281 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     2341 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttgggctcaa
     2401 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     2461 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     2521 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     2581 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     2641 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     2701 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     2761 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     2821 agtgagctga taccgctcgg tctctcatct tcaatgttgt gtctaatttt gaagttagct
     2881 ttgataccat tcttttgttt gtcagcagtg atgtaaacat tgtgagagtt atagttgtat
     2941 tccaatttgt gacctaaaat gttaccatct tctttaaaat caataccttt taattcgatt
     3001 ctattaacta aggtatcacc ttcaaacttg acttcagctc tggtcttgta gttaccgtca
     3061 tctttgaaaa aaatagttct ttcttgaaca taaccttctg gcatggcaga cttgaaaaag
     3121 tcatgttgtt tcatatgatc tgggtatcta gcaaaacatt gcaaaccata acctaaagta
     3181 gtgactaagg ttggccatgg aactggcaat ttaccagtag tacaaatcaa ttttaaggtc
     3241 aatttaccgt aagtagcatc accttcacct tcaccggaga cagaaaattt gtgaccatta
     3301 acatcaccat ctaattcaac caaaa
//
