LOCUS       sequence-128226-        2905 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gltA1-gltA2 targeting guide RNA E. coli , Low Phosphate Induction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2905)
  AUTHORS   Ye Z, Li S, Hennigan JN, Lebeau J, Moreb EA, Wolf J, Lynch MD
  TITLE     Two-stage dynamic deregulation of metabolism improves process 
  JOURNAL   Metab Eng. 2021 Nov;68:106-118. doi: 10.1016/j.ymben.2021.09.009. 
            Epub 2021 Sep 30.
  PUBMED    34600151
REFERENCE   2  (bases 1 to 2905)
  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: "2021-11"; volume: "68:106-118. doi"; pages: " 
            10.1016/j.ymben.2021.09.009. Epub 2021 Sep 30"
COMMENT     Sequence Label: pCASCADE-gltA1-gltA2
FEATURES             Location/Qualifiers
     source          1..2905
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(7..633)
                     /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     538..557
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      642..713
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      729..756
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1212..1229)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1346..1891)
                     /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(1426..1445)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2005..2099)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2120..2779)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2694..2713
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2780..2882)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 gacgtcttaa gacccacttt cacatttaag ttgtttttct aatccgcata tgatcaattc
       61 aaggccgaat aagaaggctg gctctgcacc ttggtgatca aataattcga tagcttgtcg
      121 taataatggc ggcatactat cagtagtagg tgtttccctt tcttctttag cgacttgatg
      181 ctcttgatct tccaatacgc aacctaaagt aaaatgcccc acagcgctga gtgcatataa
      241 tgcattctct agtgaaaaac cttgttggca taaaaaggct aattgatttt cgagagtttc
      301 atactgtttt tctgtaggcc gtgtacctaa atgtactttt gctccatcgc gatgacttag
      361 taaagcacat ctaaaacttt tagcgttatt acgtaaaaaa tcttgccagc tttccccttc
      421 taaagggcaa aagtgagtat ggtgcctatc taacatctca atggctaagg cgtcgagcaa
      481 agcccgctta ttttttacat gccaatacaa tgtaggctgc tctacaccta gcttctgggc
      541 gagtttacgg gttgttaaac cttcgattcc gacctcatta agcagctcta atgcgctgtt
      601 aatcacttta cttttatcta atctagacat catccaggca tcaaataaaa cgaaaggctc
      661 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctactaga
      721 gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta tacacagcta acaccacgtc
      781 gtccctatct gctgccctag gtctatgagt ggttgctgga taactctttc tgacacctta
      841 ctatcttaca aatgtaacaa aaaagttatt tttctgtaat tcgagcatgt catgttaccc
      901 cgcgagcata aaacgcgtat attcagggag accacaacgg tttccctcta caaataattt
      961 tgtttaactt tgaattcaaa agatctggta ccacctcgag ttccccgcgc cagcggggat
     1021 aaaccgaaaa gcatataatg cgtaaaagtt atgaagttcg agttccccgc gccagcgggg
     1081 ataaaccgta ttgaccaatt cattcgggac agttattagt tcgagttccc cgcgccagcg
     1141 gggataaacc gaaaaaaaaa ccccgcccct gacagggcgg ggtttttttt cctagggata
     1201 tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa
     1261 atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa
     1321 gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg
     1381 aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt
     1441 ttccccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca
     1501 ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca
     1561 gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta
     1621 tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca
     1681 gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac
     1741 tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt
     1801 ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca
     1861 agagattacg cgcagaccaa aacgatctca agaagatcat cttattaatc agataaaata
     1921 tttctagatt tcagtgcaat ttatctcttc aaatgtagca cctgaagtca gccccatacg
     1981 atataagttg ttactagtgc ttggattctc accaataaaa aacgcccggc ggcaaccgag
     2041 cgttctgaac aaatccagat ggagttctga ggtcattact ggatctatca acaggagtcc
     2101 aagcgagctc gatatcaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt
     2161 cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca
     2221 gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga
     2281 agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg
     2341 ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt
     2401 aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac
     2461 tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac
     2521 tatcccatat caccagctca ccgtctttca ttgccatacg aaattccgga tgagcattca
     2581 tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg
     2641 tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg
     2701 actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc
     2761 cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
     2821 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat
     2881 caacgtctca ttttcgccag atatc
//
