LOCUS       sequence-111971-        2780 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control guide RNA plasmid E. coli , low phosphate induction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2780)
  TITLE     SUPERSEDE
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 2780)
  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            "Unpublished"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
FEATURES             Location/Qualifiers
     source          1..2780
                     /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(1087..1104)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1221..1766)
                     /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(1301..1320)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1880..1974)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1995..2654)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2569..2588
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2655..2757)
                     /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 aaaaaccccg cccctgacag ggcggggttt tttttcctag ggatatattc
     1081 cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc
     1141 ttacgaacgg ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag
     1201 agggccgcgg caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc
     1261 tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     1321 cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg
     1381 ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt aggcagttcg
     1441 ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg
     1501 taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg cagcagccac
     1561 tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc taaactgaaa
     1621 ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa gagttggtag
     1681 ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca gagcaagaga
     1741 ttacgcgcag accaaaacga tctcaagaag atcatcttat taatcagata aaatatttct
     1801 agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata
     1861 agttgttact agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc
     1921 tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagcg
     1981 agctcgatat caaattacgc cccgccctgc cactcatcgc agtactgttg taattcatta
     2041 agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc
     2101 atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag
     2161 ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag
     2221 acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac
     2281 gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag
     2341 agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc
     2401 catatcacca gctcaccgtc tttcattgcc atacgaaatt ccggatgagc attcatcagg
     2461 cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt
     2521 aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga
     2581 aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg
     2641 atttttttct ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg
     2701 cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaacg
     2761 tctcattttc gccagatatc
//
