LOCUS       sequence-205430-        2937 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of PIS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2937)
  AUTHORS   Botero S, Chiaroni-Clarke R, Simon SM
  TITLE     Escherichia coli as a platform for the study of phosphoinositide 
            biology.
  JOURNAL   Sci Adv. 2019 Mar 27;5(3):eaat4872. doi: 10.1126/sciadv.aat4872. 
            eCollection 2019 Mar.
  PUBMED    30944849
REFERENCE   2  (bases 1 to 2937)
  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: "Sci Adv."; 
            date: "2019-03-27"; volume: "5(3):eaat4872. doi"; pages: " 
            10.1126/sciadv.aat4872. eCollection 2019 Mar"
COMMENT     Sequence Label: p15aC-1D
FEATURES             Location/Qualifiers
     source          1..2937
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     173..191
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(340..357)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(474..1019)
                     /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(554..573)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     RBS             1295..1303
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             1311..1340
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(2278..2937)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2852..2871
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
      181 tatcacgagg ccctttcgtc acacagctaa caccacgtcg tccctatctg ctgccctagg
      241 ttcagcactc ttctctttag atctagcttc atgtggcagg agaaaaaagg ctgcaccggt
      301 gcgtcagcag aatatgtgat acaggatata ttccgcttcc tcgctcactg actcgctacg
      361 ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa cggggcggag atttcctgga
      421 agatgccagg aagatactta acagggaagt gagagggccg cggcaaagcc gtttttccat
      481 aggctccgcc cccctgacaa gcatcacgaa atctgacgct caaatcagtg gtggcgaaac
      541 ccgacaggac tataaagata ccaggcgttt ccccctggcg gctccctcgt gcgctctcct
      601 gttcctgcct ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc
      661 acgcctgaca ctcagttccg ggtaggcagt tcgctccaag ctggactgta tgcacgaacc
      721 ccccgttcag tccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccgga
      781 aagacatgca aaagcaccac tggcagcagc cactggtaat tgatttagag gagttagtct
      841 tgaagtcatg cgccggttaa ggctaaactg aaaggacaag ttttggtgac tgcgctcctc
      901 caagccagtt acctcggttc aaagagttgg tagctcagag aaccttcgaa aaaccgccct
      961 gcaaggcggt tttttcgttt tcagagcaag agattacgcg cagaccaaaa cgatctcaag
     1021 aagatcatct tattaatcag ataaaatatt tctagatttc agtgcaattt atctcttcaa
     1081 atgtagcacc tgaagtcagc cccatacgat ataagttgta gcggccgcac acagctaaca
     1141 ccacgtcgtc cctatctgct gccctaggtc tatgagtggt tgctggataa ctttacgggc
     1201 atgcataagg ctcgtataat atattcaggg agaccacaac ggtttccctc tacaaataat
     1261 tttgtttaac tttgagggac tacaaaagta caagtaagga ggtatacatg gagcagaaat
     1321 tgattagcga ggaagatttg ggtagcgacg gtagcgccgg tccgaaagct aaaactacca
     1381 caccattgca ggtatgtttg ttttacccta acctcattgg ctacacacgc gttattcttt
     1441 cattgatttc cttctgcttg ctcggccact ttcccattgc atttttattt tgttacatca
     1501 ttggttttgt gctggacgcg gtggatggca tggtagcgcg acggtttggc caatgcacac
     1561 agtttggtgc aatacttgat atgttgacgg atcgggcgtc aacggctggt ttgattgttg
     1621 tggttgtgca ggtattacaa ccgcttccac attggggagc cacttcgctt gcatgtcttg
     1681 ttttcctgga tatttcttcg catttctgcg tcatgtacgt gtcgctttat gctggccgta
     1741 ctagtcataa ggacgtgagc agcagtattt tttcacttct gcgactttat tatacaaatc
     1801 gtccgtttat gtgtgcactt tgcgtcggac aggaactttt ctacttaaac ttatatatgt
     1861 acggcgtgta tgggatcgct gaacctgcgt ttctgttctt tatggctttg acagccgcgt
     1921 tgagtgcttt caagcaggtg gtgaatgtgc aacagctgct ggatagcatg tatcatctgg
     1981 cggttctgga tgctgcggga agccgccagt aatgatgagt taataacagg cctgctggta
     2041 atcgcaggcc tttttatttt tttttttagc tagcagcaga ttacgcgcag aaaaaaagga
     2101 tctcaagaag atcctttgat cttattctac ggggtctgac gctcagtgga acgaaaactc
     2161 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2221 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2281 cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg
     2341 gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc
     2401 ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac
     2461 gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc
     2521 aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata
     2581 tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc
     2641 agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc
     2701 gtctttcatt gccatacgga actccggatg agcattcatc aggcgggcaa gaatgtgaat
     2761 aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc
     2821 cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc
     2881 tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt tctccat
//
