LOCUS       sequence-311791-        2902 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low phosphate inducible gRNA array to silence the xylA and zwf 
            promoters.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2902)
  AUTHORS   Li S, Ye Z, Moreb EA, Hennigan JN, Castellanos DB, Yang T, Lynch MD
  TITLE     Dynamic control over feedback regulatory mechanisms improves NADPH 
            flux and xylitol biosynthesis in engineered E. coli.
  JOURNAL   Metab Eng. 2021 Jan 15. pii: S1096-7176(21)00013-6. doi: 
            10.1016/j.ymben.2021.01.005.
  PUBMED    33460820
REFERENCE   2  (bases 1 to 2902)
  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. 
            2021 Jan 15. pii: S1096-7176(21)00013-6. doi: 
            10.1016/j.ymben.2021.01.005."
COMMENT     Sequence Label: pCASCADE-zwf-xylA
FEATURES             Location/Qualifiers
     source          1..2902
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     129..148
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(215..317)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             complement(347..973)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     878..897
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      982..1053
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1069..1096
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1549..1566)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1683..2228)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1763..1782)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2342..2436)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2457..2902,1..214))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct
       61 tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg
      121 tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat
      181 caacggtggt atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc
      241 tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac
      301 ctcttacgtg ccgatcaacg tctcattttc gccagatatc gacgtcttaa gacccacttt
      361 cacatttaag ttgtttttct aatccgcata tgatcaattc aaggccgaat aagaaggctg
      421 gctctgcacc ttggtgatca aataattcga tagcttgtcg taataatggc ggcatactat
      481 cagtagtagg tgtttccctt tcttctttag cgacttgatg ctcttgatct tccaatacgc
      541 aacctaaagt aaaatgcccc acagcgctga gtgcatataa tgcattctct agtgaaaaac
      601 cttgttggca taaaaaggct aattgatttt cgagagtttc atactgtttt tctgtaggcc
      661 gtgtacctaa atgtactttt gctccatcgc gatgacttag taaagcacat ctaaaacttt
      721 tagcgttatt acgtaaaaaa tcttgccagc tttccccttc taaagggcaa aagtgagtat
      781 ggtgcctatc taacatctca atggctaagg cgtcgagcaa agcccgctta ttttttacat
      841 gccaatacaa tgtaggctgc tctacaccta gcttctgggc gagtttacgg gttgttaaac
      901 cttcgattcc gacctcatta agcagctcta atgcgctgtt aatcacttta cttttatcta
      961 atctagacat catccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     1021 cgttttatct gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg
     1081 ggtgggcctt tctgcgttta tacacagcta acaccacgtc gtccctatct gctgccctag
     1141 gtctatgagt ggttgctgga taactatttc tgacacctta ctatcttaca aatgtaacaa
     1201 aaaagttatt tttctgtaat tcgagcatgt catgttaccc cgcgagcata aaacgcgtat
     1261 attcagggag accacaacgg tttccctcta caaataattt tgtttaactt tgaattcaaa
     1321 agatctggta ccacctcgag ttccccgcgc cagcggggat aaaccgctcg taaaagcagt
     1381 acagtgcacc gtaagatcga gttccccgcg ccagcgggga taaaccggga gtgcccaata
     1441 ttacgacatc atccatctcg agttccccgc gccagcgggg ataaaccgaa aaaaaaaccc
     1501 cgcccctgac agggcggggt tttttttcct agggatatat tccgcttcct cgctcactga
     1561 ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg gcttacgaac ggggcggaga
     1621 tttcctggaa gatgccagga agatacttaa cagggaagtg agagggccgc ggcaaagccg
     1681 tttttccata ggctccgccc ccctgacaag catcacgaaa tctgacgctc aaatcagtgg
     1741 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggcgg ctccctcgtg
     1801 cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg
     1861 tctcattcca cgcctgacac tcagttccgg gtaggcagtt cgctccaagc tggactgtat
     1921 gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     1981 caacccggaa agacatgcaa aagcaccact ggcagcagcc actggtaatt gatttagagg
     2041 agttagtctt gaagtcatgc gccggttaag gctaaactga aaggacaagt tttggtgact
     2101 gcgctcctcc aagccagtta cctcggttca aagagttggt agctcagaga accttcgaaa
     2161 aaccgccctg caaggcggtt ttttcgtttt cagagcaaga gattacgcgc agaccaaaac
     2221 gatctcaaga agatcatctt attaatcaga taaaatattt ctagatttca gtgcaattta
     2281 tctcttcaaa tgtagcacct gaagtcagcc ccatacgata taagttgtta ctagtgcttg
     2341 gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga
     2401 gttctgaggt cattactgga tctatcaaca ggagtccaag cgagctcgat atcaaattac
     2461 gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg
     2521 aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct
     2581 tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg
     2641 tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca
     2701 ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat
     2761 atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca
     2821 gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg
     2881 tctttcattg ccatacgaaa tt
//
