LOCUS       sequence-111972-        2843 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gltA targeting guide RNA E. coli , Low Phosphate Induction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2843)
  TITLE     SUPERSEDE
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 2843)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pCASCADE-gltA2
FEATURES             Location/Qualifiers
     source          1..2843
                     /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(1150..1167)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1284..1829)
                     /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(1364..1383)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1943..2037)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2058..2717)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2632..2651
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2718..2820)
                     /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 aaaccgtatt gaccaattca ttcgggacag ttattagttc gagttccccg cgccagcggg
     1081 gataaaccga aaaaaaaacc ccgcccctga cagggcgggg ttttttttcc tagggatata
     1141 ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggaaat
     1201 ggcttacgaa cggggcggag atttcctgga agatgccagg aagatactta acagggaagt
     1261 gagagggccg cggcaaagcc gtttttccat aggctccgcc cccctgacaa gcatcacgaa
     1321 atctgacgct caaatcagtg gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     1381 ccccctggcg gctccctcgt gcgctctcct gttcctgcct ttcggtttac cggtgtcatt
     1441 ccgctgttat ggccgcgttt gtctcattcc acgcctgaca ctcagttccg ggtaggcagt
     1501 tcgctccaag ctggactgta tgcacgaacc ccccgttcag tccgaccgct gcgccttatc
     1561 cggtaactat cgtcttgagt ccaacccgga aagacatgca aaagcaccac tggcagcagc
     1621 cactggtaat tgatttagag gagttagtct tgaagtcatg cgccggttaa ggctaaactg
     1681 aaaggacaag ttttggtgac tgcgctcctc caagccagtt acctcggttc aaagagttgg
     1741 tagctcagag aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt tcagagcaag
     1801 agattacgcg cagaccaaaa cgatctcaag aagatcatct tattaatcag ataaaatatt
     1861 tctagatttc agtgcaattt atctcttcaa atgtagcacc tgaagtcagc cccatacgat
     1921 ataagttgtt actagtgctt ggattctcac caataaaaaa cgcccggcgg caaccgagcg
     1981 ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa
     2041 gcgagctcga tatcaaatta cgccccgccc tgccactcat cgcagtactg ttgtaattca
     2101 ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc
     2161 ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag
     2221 aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca gggattggct
     2281 gagacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa
     2341 cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc
     2401 cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta
     2461 tcccatatca ccagctcacc gtctttcatt gccatacgaa attccggatg agcattcatc
     2521 aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc
     2581 tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac
     2641 tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca
     2701 gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat
     2761 acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgccgatca
     2821 acgtctcatt ttcgccagat atc
//
