LOCUS       sequence-214008-        4678 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of PIS and PI4P5K.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4678)
  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 4678)
  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-5
FEATURES             Location/Qualifiers
     source          1..4678
                     /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"
     RBS             2031..2039
                     /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             2051..2080
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(4019..4678)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4593..4612
                     /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 aatcgcacta gccgctatat taaggaggta
     2041 ataaatggtg gaacaaaagt tgatcagcga agaagacttg ggcagcgatg gtagcgcggg
     2101 tgcgtcggcc tcctccgggc cgtcgtcttc ggtcggtttt tcatcctttg atcccgcggt
     2161 cccttcctgt accttgtcct cagcatctgg aatcaagaga cccatggcat ctgaggtgcc
     2221 ttatgcctct ggcatgccca tcaagaaaat aggccataga agtgttgatt cctcaggaga
     2281 gacaacatat aaaaagacaa cctcatcagc cttgaaaggt gccatccagt taggcattac
     2341 ccacactgtg gggagcctga gtaccaaacc agagcgtgat gtcctcatgc aagatttcta
     2401 cgtggttgag agtatcttct ttcccagtga agggagcaac ctgacccctg ctcatcacta
     2461 caatgacttt cgtttcaaga cctatgcacc tgttgccttc cgctacttcc gggagctatt
     2521 tggtatccgg cccgatgatt acttgtattc cctctgcagt gagccgctga ttgaactctg
     2581 tagctctgga gctagtggtt ccctattcta tgtgtccagc gacgatgagt tcattattaa
     2641 gacagtccaa cataaagagg cggaatttct gcagaagctg cttccaggat actacatgaa
     2701 cctcaaccag aaccctcgga ctttgctgcc taaattctat ggactgtact gtgtgcaggc
     2761 aggtggcaag aacattcgga ttgtggtgat gaacaatctt ttaccaagat cggtaaaaat
     2821 gcatatcaaa tatgacctca aaggctcaac ctacaaacgg cgggcttccc agaaagagcg
     2881 agagaagcct cttcccacat ttaaagacct agacttctta caagacatcc ctgatggtct
     2941 ttttttggat gctgacatgt acaacgctct ctgtaagacc ctgcagcgtg actgtttggt
     3001 gctgcagagc ttcaagataa tggattacag cctcttgatg tcaatccata atatagatca
     3061 tgcacaacga gagcccttaa gcagtgaaac acagtactca gttgatactc gaagaccggc
     3121 cccccaaaag gctctgtatt ccacagccat ggaatccatc cagggagagg ctcgacgggg
     3181 tggtaccatg gagactgatg accatatggg tggcatccct gcccggaata gtaaagggga
     3241 aaggcttctg ctttatattg gcatcattga cattctacag tcttacaggt ttgttaagaa
     3301 gttggagcac tcttggaaag ccctggtaca tgacggagac actgtctcag tgcatcgccc
     3361 aggcttctac gctgaacggt tccagcgctt catgtgcaac acagtattta agaagattcc
     3421 cttgaagcct tctccttcca aaaagtttcg gtctggctca tctttctctc ggcgagcagg
     3481 ctccagtggc aactcctgca ttacttacca gccatcggtc tctggggaac acaaggcaca
     3541 agtgacaaca aaggcagaag tggagccagg cgttcacctt ggtcgtcctg atgttttacc
     3601 tcagactcca cctttggagg aaatcagtga gggctcgcct attcctgacc ccagtttctc
     3661 acctctagtt ggagagactt tgcaaatgct aactacaagt acaaccttgg aaaagcttga
     3721 agttgcagag tcagagttca cccattaata ataatgatga gttaataaca gcctgctggt
     3781 aatcgcaggc ctttttattt ttttttttag ctagcagcag attacgcgca gaaaaaaagg
     3841 atctcaagaa gatcctttga tcttattcta cggggtctga cgctcagtgg aacgaaaact
     3901 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3961 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4021 acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat
     4081 ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc
     4141 cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca
     4201 cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct
     4261 caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat
     4321 atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt
     4381 cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac
     4441 cgtctttcat tgccatacgg aactccggat gagcattcat caggcgggca agaatgtgaa
     4501 taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat
     4561 ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt
     4621 ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccat
//
