LOCUS       sequence-174939-        2966 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gltA1-gltA2-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 2966)
  TITLE     SUPERSEDE
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 2966)
  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            "Unpublished"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-GapAP1
FEATURES             Location/Qualifiers
     source          1..2966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(6..632)
                     /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     537..556
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      641..712
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      728..755
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1272..1289)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1406..1951)
                     /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(1486..1505)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2065..2159)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2180..2839)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2754..2773
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2840..2942)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 acgtcttaag acccactttc acatttaagt tgtttttcta atccgcatat gatcaattca
       61 aggccgaata agaaggctgg ctctgcacct tggtgatcaa ataattcgat agcttgtcgt
      121 aataatggcg gcatactatc agtagtaggt gtttcccttt cttctttagc gacttgatgc
      181 tcttgatctt ccaatacgca acctaaagta aaatgcccca cagcgctgag tgcatataat
      241 gcattctcta gtgaaaaacc ttgttggcat aaaaaggcta attgattttc gagagtttca
      301 tactgttttt ctgtaggccg tgtacctaaa tgtacttttg ctccatcgcg atgacttagt
      361 aaagcacatc taaaactttt agcgttatta cgtaaaaaat cttgccagct ttccccttct
      421 aaagggcaaa agtgagtatg gtgcctatct aacatctcaa tggctaaggc gtcgagcaaa
      481 gcccgcttat tttttacatg ccaatacaat gtaggctgct ctacacctag cttctgggcg
      541 agtttacggg ttgttaaacc ttcgattccg acctcattaa gcagctctaa tgcgctgtta
      601 atcactttac ttttatctaa tctagacatc atccaggcat caaataaaac gaaaggctca
      661 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag
      721 tcacactggc tcaccttcgg gtgggccttt ctgcgtttat acacagctaa caccacgtcg
      781 tccctatctg ctgccctagg tctatgagtg gttgctggat aactctttct gacaccttac
      841 tatcttacaa atgtaacaaa aaagttattt ttctgtaatt cgagcatgtc atgttacccc
      901 gcgagcataa aacgcgtata ttcagggaga ccacaacggt ttccctctac aaataatttt
      961 gtttaacttt gaattcaaaa gatctggtac cacctcgagt tccccgcgcc agcggggata
     1021 aaccgaaaag catataatgc gtaaaagtta tgaagttcga gttccccgcg ccagcgggga
     1081 taaaccgtat tgaccaattc attcgggaca gttattagtt cgagttcccc gcgccagcgg
     1141 ggataaaccg gtttttgtaa ttttacaggc aaccttttat tcgagttccc cgcgccagcg
     1201 gggataaacc gaaaaaaaaa ccccgcccct gacagggcgg ggtttttttt cctagggata
     1261 tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa
     1321 atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa
     1381 gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg
     1441 aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt
     1501 ttccccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca
     1561 ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca
     1621 gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta
     1681 tccggtaact atcgtcttga gtccaacccg gaaagacata caaaagcacc actggcagca
     1741 gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac
     1801 tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt
     1861 ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca
     1921 agagattacg cgcagaccaa aacgatctca agaagatcat cttattaatc agataaaata
     1981 tttctagatt tcagtgcaat ttatctcttc aaatgtagca cctgaagtca gccccatacg
     2041 atataagttg ttactagtgc ttggattctc accaataaaa aacgcccggc ggcaaccgag
     2101 cgttctgaac aaatccagat ggagttctga ggtcattact ggatctatca acaggagtcc
     2161 aagcgagctc gatatcaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt
     2221 cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca
     2281 gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga
     2341 agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg
     2401 ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt
     2461 aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac
     2521 tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac
     2581 tatcccatat caccagctca ccgtctttca ttgccatacg aaattccgga tgagcattca
     2641 tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg
     2701 tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg
     2761 actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc
     2821 cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
     2881 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat
     2941 caacgtctca ttttcgccag atatcg
//
