LOCUS       sequence-174934-        3149 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fabI-gltA1-gltA2-udhA-zwf-gapAP1 targeting guide RNA E. coli, Low 
            Phosphate Induction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3149)
  TITLE     SUPERSEDE
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3149)
  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            "Unpublished"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-fabI-gltA1-gltA2-udhA-zwf-gapAP1
FEATURES             Location/Qualifiers
     source          1..3149
                     /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(1456..1473)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1590..2135)
                     /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(1670..1689)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2249..2343)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2364..3023)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2938..2957
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3024..3126)
                     /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 aaaccgttga ttataataac cgtttatctg ttcgtatcga gttccccgcg ccagcgggga
     1081 taaaccgaaa agcatataat gcgtaaaagt tatgaagttc gagttccccg cgccagcggg
     1141 gataaaccgt attgaccaat tcattcggga cagttattag ttcgagttcc ccgcgccagc
     1201 ggggataaac cgttaccatt ctgttgcttt tatgtataag aatcgagttc cccgcgccag
     1261 cggggataaa ccgctcgtaa aagcagtaca gtgcaccgta agatcgagtt ccccgcgcca
     1321 gcggggataa accggttttt gtaattttac aggcaacctt ttattcgagt tccccgcgcc
     1381 agcggggata aaccgaaaaa aaaaccccgc ccctgacagg gcggggtttt ttttcctagg
     1441 gatatattcc gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc
     1501 ggaaatggct tacgaacggg gcggagattt cctggaagat gccaggaaga tacttaacag
     1561 ggaagtgaga gggccgcggc aaagccgttt ttccataggc tccgcccccc tgacaagcat
     1621 cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga caggactata aagataccag
     1681 gcgtttcccc ctggcggctc cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt
     1741 gtcattccgc tgttatggcc gcgtttgtct cattccacgc ctgacactca gttccgggta
     1801 ggcagttcgc tccaagctgg actgtatgca cgaacccccc gttcagtccg accgctgcgc
     1861 cttatccggt aactatcgtc ttgagtccaa cccggaaaga catacaaaag caccactggc
     1921 agcagccact ggtaattgat ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct
     1981 aaactgaaag gacaagtttt ggtgactgcg ctcctccaag ccagttacct cggttcaaag
     2041 agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa ggcggttttt tcgttttcag
     2101 agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga tcatcttatt aatcagataa
     2161 aatatttcta gatttcagtg caatttatct cttcaaatgt agcacctgaa gtcagcccca
     2221 tacgatataa gttgttacta gtgcttggat tctcaccaat aaaaaacgcc cggcggcaac
     2281 cgagcgttct gaacaaatcc agatggagtt ctgaggtcat tactggatct atcaacagga
     2341 gtccaagcga gctcgatatc aaattacgcc ccgccctgcc actcatcgca gtactgttgt
     2401 aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc
     2461 gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg
     2521 gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga
     2581 ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca
     2641 ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat
     2701 tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga
     2761 acactatccc atatcaccag ctcaccgtct ttcattgcca tacgaaattc cggatgagca
     2821 ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt
     2881 acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca
     2941 actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta
     3001 tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct cgataactca
     3061 aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc
     3121 cgatcaacgt ctcattttcg ccagatatc
//
