LOCUS       sequence-244769-        3065 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 3065)
  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 3065)
  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..3065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(421..753)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSILEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCRRPFICT
                     IHPYHG"
     primer_bind     648..670
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     662..679
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     663..679
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    680..736
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(749..765)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(749..765)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(762..784)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    773..789
                     /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)."
     promoter        complement(join(797..803,804..821,822..827))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    842..863
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    1032..1065
                     /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)."
     rep_origin      complement(1385..1973)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1465..1484)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2201..2860)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2775..2794
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(3023..3052)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 ggtctcatta tgttcaagaa agaactattt ttttcaaaga tgacggtaac tacaagacca
       61 gagctgaagt caagtttgaa ggtgatacct tagttaatag aatcgaatta aaaggtattg
      121 attttaaaga agatggtaac attttaggtc acaaattgga atacaactat aactctcaca
      181 atgtttacat cactgctgac aaacaaaaga atggtatcaa agctaacttc aaaattagac
      241 acaacattga agatggtggt gttcaattag ctgaccatta tcaacaaaat actccaattg
      301 gtgatggtcc agtcttgtta ccagacaacc attacttatc ctatcaatct gccttatcca
      361 aagatccaaa cgaaaagaga gatcacatgg tcttgttaga atttgttact gctgctggta
      421 ttacccatgg tatggatgaa ttgtacaaat aaaaggtctt ctgcaccata tgcggtgtga
      481 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
      541 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      601 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      661 ttgtaaaacg acggccagtg aattcgagct cggtacccgg ggatcctcta gaatcgacct
      721 gcaggcatgc aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
      781 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
      841 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtagggaa
      901 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggaagacc tcctcttgcc
      961 catcgaacgt acaagtactc ctctgttctc tccttccttt gcttcttcgt acgctgcagg
     1021 tcgacgaatt ctaccgttcg tataatgtat gctatacgaa gttatagatc tgtttagctt
     1081 gcctcgtccc cgccgggtca cccggccagc gacatggagg cccagaatac cctccttgac
     1141 agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat ttagcccata
     1201 catccccatg tataatcatt tgcatccata cattttgatg gccgcacaga gaccgagcgg
     1261 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     1321 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     1381 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     1441 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     1501 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     1561 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     1621 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     1681 gtaactatcg tcttgagccc aacccggtaa gacacgactt atcgccactg gcagcagcca
     1741 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     1801 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     1861 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     1921 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     1981 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     2041 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     2101 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagagttctg aggtcattac
     2161 tggatctatc aacagcagtc caagcgagct cgatatcaaa ttacgccccg ccctgccact
     2221 catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg
     2281 catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc
     2341 ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg
     2401 tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga
     2461 aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc
     2521 ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa
     2581 cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac
     2641 gaaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact
     2701 tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt
     2761 tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg
     2821 atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg
     2881 aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt
     2941 tggaacctct tacgtgcccg atcaactcgc gcgtttgcca cctgacgtct aagaaaagga
     3001 atattcagca atttgcccgt gccgaagaaa ggcccacccg tgaaggtgag ccagtgagtt
     3061 gattg
//
