LOCUS       sequence-174940-        3027 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gltA1-gltA2-udhA-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 3027)
  TITLE     SUPERSEDE
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3027)
  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            "Unpublished"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-gltA1-gltA2-udhA-GapAP1
FEATURES             Location/Qualifiers
     source          1..3027
                     /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(1334..1351)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1468..2013)
                     /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(1548..1567)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2127..2221)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2242..2901)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2816..2835
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2902..3004)
                     /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 gcatataatg cgtaaaagtt atgaagttcg agttccccgc gccagcgggg
     1081 ataaaccgta ttgaccaatt cattcgggac agttattagt tcgagttccc cgcgccagcg
     1141 gggataaacc gttaccattc tgttgctttt atgtataaga atcgagttcc ccgcgccagc
     1201 ggggataaac cggtttttgt aattttacag gcaacctttt attcgagttc cccgcgccag
     1261 cggggataaa ccgaaaaaaa aaccccgccc ctgacagggc ggggtttttt ttcctaggga
     1321 tatattccgc ttcctcgctc actgactcgc tacgctcggt cgttcgactg cggcgagcgg
     1381 aaatggctta cgaacggggc ggagatttcc tggaagatgc caggaagata cttaacaggg
     1441 aagtgagagg gccgcggcaa agccgttttt ccataggctc cgcccccctg acaagcatca
     1501 cgaaatctga cgctcaaatc agtggtggcg aaacccgaca ggactataaa gataccaggc
     1561 gtttccccct ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt
     1621 cattccgctg ttatggccgc gtttgtctca ttccacgcct gacactcagt tccgggtagg
     1681 cagttcgctc caagctggac tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct
     1741 tatccggtaa ctatcgtctt gagtccaacc cggaaagaca tacaaaagca ccactggcag
     1801 cagccactgg taattgattt agaggagtta gtcttgaagt catgcgccgg ttaaggctaa
     1861 actgaaagga caagttttgg tgactgcgct cctccaagcc agttacctcg gttcaaagag
     1921 ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg cggttttttc gttttcagag
     1981 caagagatta cgcgcagacc aaaacgatct caagaagatc atcttattaa tcagataaaa
     2041 tatttctaga tttcagtgca atttatctct tcaaatgtag cacctgaagt cagccccata
     2101 cgatataagt tgttactagt gcttggattc tcaccaataa aaaacgcccg gcggcaaccg
     2161 agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt
     2221 ccaagcgagc tcgatatcaa attacgcccc gccctgccac tcatcgcagt actgttgtaa
     2281 ttcattaagc attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc
     2341 cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc
     2401 gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt
     2461 ggctgagacg aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc
     2521 gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc
     2581 actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac
     2641 actatcccat atcaccagct caccgtcttt cattgccata cgaaattccg gatgagcatt
     2701 catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac
     2761 ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac
     2821 tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata
     2881 tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
     2941 aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccg
     3001 atcaacgtct cattttcgcc agatatc
//
