LOCUS       sequence-254087-        3066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrative yeast terminator characterization plasmid. Clone in with
            BpiI. Has mVenus fragment upstream and a KanMX fragment downstream..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3066)
  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 3066)
  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: pEMY01CD-TER
FEATURES             Location/Qualifiers
     source          1..3066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    117..138
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(153..158,159..176,177..183)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    191..207
                     /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     196..218
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     215..231
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     215..231
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             227..559
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSILEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISRRIWCRRPFICT
                     IHPYHG"
     misc_feature    244..300
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(301..318)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(301..317)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(310..332)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      993..1022
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     CDS             1185..1844
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1251..1270)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2072..2660
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2561..2580
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(2980..3013)
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 agtacttgta cgttcgatgg gcaagaggag gtcttccccg cgcgttggcc gattcattaa
       61 tgcagctggc acgacaggtt tccctactgg aaagcgggca gtgagcgcaa cgcaattaat
      121 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
      181 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
      241 gccaagcttg catgcctgca ggtcgattct agaggatccc cgggtaccga gctcgaattc
      301 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
      361 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
      421 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct
      481 tacgcatctg tgcggtattt cacgccgcat atggtgcaga agacctttta tttgtacaat
      541 tcatccatac catgggtaat accagcagca gtaacaaatt ctaacaagac catgtgatct
      601 ctcttttcgt ttggatcttt ggataaggca gattgatagg ataagtaatg gttgtctggt
      661 aacaagactg gaccatcacc aattggagta ttttgttgat aatggtcagc taattgaaca
      721 ccaccatctt caatgttgtg tctaattttg aagttagctt tgataccatt cttttgtttg
      781 tcagcagtga tgtaaacatt gtgagagtta tagttgtatt ccaatttgtg acctaaaatg
      841 ttaccatctt ctttaaaatc aatacctttt aattcgattc tattaactaa ggtatcacct
      901 tcaaacttga cttcagctct ggtcttgtag ttaccgtcat ctttgaaaaa aatagttctt
      961 tcttgaacat aatgagaccc aatcaactca ctggctcacc ttcacgggtg ggcctttctt
     1021 cggcacgggc aaattgctga atattccttt tcttagacgt caggtggcaa acgcgcgagt
     1081 tgatcgggca cgtaagaggt tccaactttc accataatga aataagatca ctaccgggcg
     1141 tattttttga gttatcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca
     1201 ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc
     1261 agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa
     1321 agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc
     1381 tgatgaatgc tcatccggaa tttcgtatgg caatgaaaga cggtgagctg gtgatatggg
     1441 atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct
     1501 ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt
     1561 gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct
     1621 cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact
     1681 tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc
     1741 cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta
     1801 atgaattaca acagtactgc gatgagtggc agggcggggc gtaatttgat atcgagctcg
     1861 cttggactgc tgttgataga tccagtaatg acctcagaac tctgtcagac caagtttact
     1921 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1981 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     2041 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2101 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     2161 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     2221 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     2281 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     2341 ggttgggctc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     2401 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     2461 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     2521 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     2581 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     2641 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     2701 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     2761 ttaccgcctt tgagtgagct gataccgctc ggtctctgtg cggccatcaa aatgtatgga
     2821 tgcaaatgat tatacatggg gatgtatggg ctaaatgtac gggcgacagt cacatcatgc
     2881 ccctgagctg cgcacgtcaa gactgtcaag gagggtattc tgggcctcca tgtcgctggc
     2941 cgggtgaccc ggcggggacg aggcaagcta aacagatcta taacttcgta tagcatacat
     3001 tatacgaacg gtagaattcg tcgacctgca gcgtacgaag aaagcaaagg aaggagagaa
     3061 cagagg
//
