LOCUS       sequence-174933-        2902 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  udhA-zwf targeting guide RNA E. coli , Low Phosphate Induction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2902)
  TITLE     SUPERSEDE
  JOURNAL   Unpublished
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: 
            "Unpublished"
COMMENT     Sequence Label: pCASCADE-udhA-zwf
FEATURES             Location/Qualifiers
     source          1..2902
                     /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(1209..1226)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1343..1888)
                     /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(1423..1442)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2002..2096)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2117..2776)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2691..2710
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2777..2879)
                     /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 aaaccgttac cattctgttg cttttatgta taagaatcga gttccccgcg ccagcgggga
     1081 taaaccgctc gtaaaagcag tacagtgcac cgtaagatcg agttccccgc gccagcgggg
     1141 ataaaccgaa aaaaaaaccc cgcccctgac agggcggggt tttttttcct agggatatat
     1201 tccgcttcct cgctcactga ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg
     1261 gcttacgaac ggggcggaga tttcctggaa gatgccagga agatacttaa cagggaagtg
     1321 agagggccgc ggcaaagccg tttttccata ggctccgccc ccctgacaag catcacgaaa
     1381 tctgacgctc aaatcagtgg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1441 cccctggcgg ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc
     1501 cgctgttatg gccgcgtttg tctcattcca cgcctgacac tcagttccgg gtaggcagtt
     1561 cgctccaagc tggactgtat gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc
     1621 ggtaactatc gtcttgagtc caacccggaa agacatacaa aagcaccact ggcagcagcc
     1681 actggtaatt gatttagagg agttagtctt gaagtcatgc gccggttaag gctaaactga
     1741 aaggacaagt tttggtgact gcgctcctcc aagccagtta cctcggttca aagagttggt
     1801 agctcagaga accttcgaaa aaccgccctg caaggcggtt ttttcgtttt cagagcaaga
     1861 gattacgcgc agaccaaaac gatctcaaga agatcatctt attaatcaga taaaatattt
     1921 ctagatttca gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc ccatacgata
     1981 taagttgtta ctagtgcttg gattctcacc aataaaaaac gcccggcggc aaccgagcgt
     2041 tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca ggagtccaag
     2101 cgagctcgat atcaaattac gccccgccct gccactcatc gcagtactgt tgtaattcat
     2161 taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg
     2221 gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga
     2281 agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg
     2341 agacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac
     2401 acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc
     2461 agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat
     2521 cccatatcac cagctcaccg tctttcattg ccatacgaaa ttccggatga gcattcatca
     2581 ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct
     2641 ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact
     2701 gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag
     2761 tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata
     2821 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa
     2881 cgtctcattt tcgccagata tc
//
