LOCUS       sequence-214011-        7316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of PIS, PI4K and PI4P5K.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7316)
  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 7316)
  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-4D1D-5
FEATURES             Location/Qualifiers
     source          1..7316
                     /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"
     RBS             365..373
                     /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             382..411
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2978..2995)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3112..3657)
                     /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(3192..3211)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     RBS             3933..3941
                     /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             3949..3978
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     RBS             4669..4677
                     /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             4689..4718
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(6657..7316)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7231..7250
                     /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 tctatgagtg gttgctggat aactttacgg gcatgcataa ggctcgtata atatattcag
      301 ggagaccaca acggtttccc tctacaaata attttgttta actttacaac acgaacgaaa
      361 aacataagga ggtacataat ggaacagaaa ttgattagcg aagaggattt gggtagcgat
      421 ggcagcgcag gcggagacac gatcgttgaa ccggcccccc tgaagccaac ttctgagccc
      481 gctcccggcc caccagggaa taatggtggc tccttgctaa gtgtcatcac ggagggggtc
      541 ggggagctct cggtgattga ccctgaggtg gcccagaagg cctgccagga ggtgctggag
      601 aaagtcaagc tcttgcatgg aggcgtggcc atctctagca gaggcacccc cctggagctg
      661 gtcaatggag acggagtaga cagcgagatc cgttgcctgg acgacccacc cgcccagata
      721 agggaggagg aagatgagat gggggccacg gtggcctcag gcacagccaa gggagcaaga
      781 aggcggcggc agaacaactc ggccaaacag tcttggctgc tgaggctgtt tgagtcgaaa
      841 ctgtttgaca tctccatggc catttcatac ctgtataact ccaaggagcc tggagtgcag
      901 gcctacattg gcaaccggct cttctgcttt cgtaatgagg atgtggactt ctatctgccc
      961 cagttgctta acatgtacat ccacatggac gaggacgtgg gtgatgccat caagccctac
     1021 atagtccacc gctgccgcca gagcattaac ttttccctcc agtgtgctct gttgctgggg
     1081 gcctactctt cagacatgca catttccact cagcgacact cccgtgggac caagctacgg
     1141 aagctgatcc tctcagatga gctgaagccc gctcaccgaa agagggagct gccctccttg
     1201 agcccagccc ccgacacagg gctgtctccc tctaaaagga cccaccagcg ctctaagtca
     1261 gatgccaccg ccagcataag tctcagcagc aacttgaaac gaacagccag caaccctaaa
     1321 gtggagaatg aggacgagcc cgtcagactg gctcctgagc gagaattcat caagtccctg
     1381 atggcgattg gcaagcggct ggccacactc cccaccaagg agcagaagac acagcggctg
     1441 atctcagagc tctccctgct caaccacaag ctccctgccc gagtctggct gcccactgct
     1501 ggcttcgacc accacgtggt ccgcgtgccc cacacccagg ctgtcgtcct caattctaag
     1561 gacaaggctc cctacctgat ctatgtggag gtcctcgaat gtgaaaactt cgacaccact
     1621 agtgtccccg cccgaatccc cgagaaccga attcggagca cccggtctgt agagaacctg
     1681 cccgaatgcg gcatcaccca cgagcagcgg gcaggcagct tcagcaccgt gcccaactat
     1741 gacaacgatg acgaggcctg gtcggtggat gacataggcg agctgcaggt ggagctccct
     1801 gaagtgcaca ccaacagctg tgacaacatc tcccagttct ccgtggacag catcaccagc
     1861 caggaaagca aggagcctgt gttcatcgca gcgggtgaca tccgacggcg cctttcagag
     1921 cagctggctc acacccccac agccttcaaa cgagacccgg aagacccttc tgcagttgct
     1981 ctcaaagagc cctggcagga gaaagtacgg cgcattagag agggttcccc ctatggccat
     2041 ctccccaatt ggcggctcct gtcagtcatc gtcaagtgtg gggatgacct tcggcaggag
     2101 ctgttggcct ttcaggtgtt aaagcaactg cagtccattt gggaacagga gcgagtaccc
     2161 ctgtggatca agccatataa gattcttgtg atttcggccg acagtggcat gattgaacca
     2221 gtggtcaacg ctgtatccat tcaccaggtg aagaagcagt cacagctctc cctgctcgat
     2281 tacttcctac aggagcatgg cagttatacc actgaggcat tcctcagcgc ccagcgcaat
     2341 tttgtgcaga gttgtgctgg ctactgcttg ggctgctacc tgctgcaggt caaggacaga
     2401 cacaatggga acatcctgtt ggacgcagaa ggccacatca tccacatcga ctttggcttc
     2461 atcctatcca gctcaccccg aaacctgggc tttgagacat cagccttcaa actgaccaca
     2521 gagtttgttg acgtgatggg cggcctggat ggcgacatgt tcaactatta caagatgttg
     2581 atgctgcagg ggctgatcgc tgctcggaaa cacatggata aggtggtgca gatcgtagag
     2641 atcatgcaac aaggttctca gcttccttgc ttccatggct ccagcaccat ccgcaacctc
     2701 aaggagaggt tccacatgag catgaccgag gagcagctgc agctgctggt ggagcagatg
     2761 gtggatggca gcatgcggtc tataaccacc aaactctatg acggcttcca gtacctcacc
     2821 aacggcatca tgtaaggatc ctgatgagtt aataacaggc ctgctggtaa tcgcaggcct
     2881 ttttattttt ttttttagat ctagcttcat gtggcaggag aaaaaaggct gcaccggtgc
     2941 gtcagcagaa tatgtgatac aggatatatt ccgcttcctc gctcactgac tcgctacgct
     3001 cggtcgttcg actgcggcga gcggaaatgg cttacgaacg gggcggagat ttcctggaag
     3061 atgccaggaa gatacttaac agggaagtga gagggccgcg gcaaagccgt ttttccatag
     3121 gctccgcccc cctgacaagc atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc
     3181 gacaggacta taaagatacc aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt
     3241 tcctgccttt cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac
     3301 gcctgacact cagttccggg taggcagttc gctccaagct ggactgtatg cacgaacccc
     3361 ccgttcagtc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggaaa
     3421 gacatgcaaa agcaccactg gcagcagcca ctggtaattg atttagagga gttagtcttg
     3481 aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt ttggtgactg cgctcctcca
     3541 agccagttac ctcggttcaa agagttggta gctcagagaa ccttcgaaaa accgccctgc
     3601 aaggcggttt tttcgttttc agagcaagag attacgcgca gaccaaaacg atctcaagaa
     3661 gatcatctta ttaatcagat aaaatatttc tagatttcag tgcaatttat ctcttcaaat
     3721 gtagcacctg aagtcagccc catacgatat aagttgtagc ggccgcacac agctaacacc
     3781 acgtcgtccc tatctgctgc cctaggtcta tgagtggttg ctggataact ttacgggcat
     3841 gcataaggct cgtataatat attcagggag accacaacgg tttccctcta caaataattt
     3901 tgtttaactt tgagggacta caaaagtaca agtaaggagg tatacatgga gcagaaattg
     3961 attagcgagg aagatttggg tagcgacggt agcgccggtc cgaaagctaa aactaccaca
     4021 ccattgcagg tatgtttgtt ttaccctaac ctcattggct acacacgcgt tattctttca
     4081 ttgatttcct tctgcttgct cggccacttt cccattgcat ttttattttg ttacatcatt
     4141 ggttttgtgc tggacgcggt ggatggcatg gtagcgcgac ggtttggcca atgcacacag
     4201 tttggtgcaa tacttgatat gttgacggat cgggcgtcaa cggctggttt gattgttgtg
     4261 gttgtgcagg tattacaacc gcttccacat tggggagcca cttcgcttgc atgtcttgtt
     4321 ttcctggata tttcttcgca tttctgcgtc atgtacgtgt cgctttatgc tggccgtact
     4381 agtcataagg acgtgagcag cagtattttt tcacttctgc gactttatta tacaaatcgt
     4441 ccgtttatgt gtgcactttg cgtcggacag gaacttttct acttaaactt atatatgtac
     4501 ggcgtgtatg ggatcgctga acctgcgttt ctgttcttta tggctttgac agccgcgttg
     4561 agtgctttca agcaggtggt gaatgtgcaa cagctgctgg atagcatgta tcatctggcg
     4621 gttctggatg ctgcgggaag ccgccagtaa tcgcactagc cgctatatta aggaggtaat
     4681 aaatggtgga acaaaagttg atcagcgaag aagacttggg cagcgatggt agcgcgggtg
     4741 cgtcggcctc ctccgggccg tcgtcttcgg tcggtttttc atcctttgat cccgcggtcc
     4801 cttcctgtac cttgtcctca gcatctggaa tcaagagacc catggcatct gaggtgcctt
     4861 atgcctctgg catgcccatc aagaaaatag gccatagaag tgttgattcc tcaggagaga
     4921 caacatataa aaagacaacc tcatcagcct tgaaaggtgc catccagtta ggcattaccc
     4981 acactgtggg gagcctgagt accaaaccag agcgtgatgt cctcatgcaa gatttctacg
     5041 tggttgagag tatcttcttt cccagtgaag ggagcaacct gacccctgct catcactaca
     5101 atgactttcg tttcaagacc tatgcacctg ttgccttccg ctacttccgg gagctatttg
     5161 gtatccggcc cgatgattac ttgtattccc tctgcagtga gccgctgatt gaactctgta
     5221 gctctggagc tagtggttcc ctattctatg tgtccagcga cgatgagttc attattaaga
     5281 cagtccaaca taaagaggcg gaatttctgc agaagctgct tccaggatac tacatgaacc
     5341 tcaaccagaa ccctcggact ttgctgccta aattctatgg actgtactgt gtgcaggcag
     5401 gtggcaagaa cattcggatt gtggtgatga acaatctttt accaagatcg gtaaaaatgc
     5461 atatcaaata tgacctcaaa ggctcaacct acaaacggcg ggcttcccag aaagagcgag
     5521 agaagcctct tcccacattt aaagacctag acttcttaca agacatccct gatggtcttt
     5581 ttttggatgc tgacatgtac aacgctctct gtaagaccct gcagcgtgac tgtttggtgc
     5641 tgcagagctt caagataatg gattacagcc tcttgatgtc aatccataat atagatcatg
     5701 cacaacgaga gcccttaagc agtgaaacac agtactcagt tgatactcga agaccggccc
     5761 cccaaaaggc tctgtattcc acagccatgg aatccatcca gggagaggct cgacggggtg
     5821 gtaccatgga gactgatgac catatgggtg gcatccctgc ccggaatagt aaaggggaaa
     5881 ggcttctgct ttatattggc atcattgaca ttctacagtc ttacaggttt gttaagaagt
     5941 tggagcactc ttggaaagcc ctggtacatg acggagacac tgtctcagtg catcgcccag
     6001 gcttctacgc tgaacggttc cagcgcttca tgtgcaacac agtatttaag aagattccct
     6061 tgaagccttc tccttccaaa aagtttcggt ctggctcatc tttctctcgg cgagcaggct
     6121 ccagtggcaa ctcctgcatt acttaccagc catcggtctc tggggaacac aaggcacaag
     6181 tgacaacaaa ggcagaagtg gagccaggcg ttcaccttgg tcgtcctgat gttttacctc
     6241 agactccacc tttggaggaa atcagtgagg gctcgcctat tcctgacccc agtttctcac
     6301 ctctagttgg agagactttg caaatgctaa ctacaagtac aaccttggaa aagcttgaag
     6361 ttgcagagtc agagttcacc cattaataat aatgatgagt taataacagc ctgctggtaa
     6421 tcgcaggcct ttttattttt ttttttagct agcagcagat tacgcgcaga aaaaaaggat
     6481 ctcaagaaga tcctttgatc ttattctacg gggtctgacg ctcagtggaa cgaaaactca
     6541 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6601 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6661 gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg
     6721 aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct
     6781 tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg
     6841 tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca
     6901 ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat
     6961 atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca
     7021 gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg
     7081 tctttcattg ccatacggaa ctccggatga gcattcatca ggcgggcaag aatgtgaata
     7141 aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc
     7201 agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct
     7261 ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt ctccat
//
