LOCUS       sequence-175306-        3088 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fabI-gltA1-gltA2-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 3088)
  TITLE     SUPERSEDE
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3088)
  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            "Unpublished"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
FEATURES             Location/Qualifiers
     source          1..3088
                     /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(1395..1412)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1529..2074)
                     /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(1609..1628)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2188..2282)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2303..2962)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2877..2896
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2963..3065)
                     /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 accgaaaaaa aaaccccgcc cctgacaggg cggggttttt tttcctaggg
     1381 atatattccg cttcctcgct cactgactcg ctacgctcgg tcgttcgact gcggcgagcg
     1441 gaaatggctt acgaacgggg cggagatttc ctggaagatg ccaggaagat acttaacagg
     1501 gaagtgagag ggccgcggca aagccgtttt tccataggct ccgcccccct gacaagcatc
     1561 acgaaatctg acgctcaaat cagtggtggc gaaacccgac aggactataa agataccagg
     1621 cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg tttaccggtg
     1681 tcattccgct gttatggccg cgtttgtctc attccacgcc tgacactcag ttccgggtag
     1741 gcagttcgct ccaagctgga ctgtatgcac gaaccccccg ttcagtccga ccgctgcgcc
     1801 ttatccggta actatcgtct tgagtccaac ccggaaagac atacaaaagc accactggca
     1861 gcagccactg gtaattgatt tagaggagtt agtcttgaag tcatgcgccg gttaaggcta
     1921 aactgaaagg acaagttttg gtgactgcgc tcctccaagc cagttacctc ggttcaaaga
     1981 gttggtagct cagagaacct tcgaaaaacc gccctgcaag gcggtttttt cgttttcaga
     2041 gcaagagatt acgcgcagac caaaacgatc tcaagaagat catcttatta atcagataaa
     2101 atatttctag atttcagtgc aatttatctc ttcaaatgta gcacctgaag tcagccccat
     2161 acgatataag ttgttactag tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc
     2221 gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag
     2281 tccaagcgag ctcgatatca aattacgccc cgccctgcca ctcatcgcag tactgttgta
     2341 attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg
     2401 ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg
     2461 cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat
     2521 tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac
     2581 cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt
     2641 cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa
     2701 cactatccca tatcaccagc tcaccgtctt tcattgccat acgaaattcc ggatgagcat
     2761 tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta
     2821 cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa
     2881 ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat
     2941 atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa
     3001 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     3061 gatcaacgtc tcattttcgc cagatatc
//
