LOCUS       sequence-213351-        5575 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of PIS and PI4K.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5575)
  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 5575)
  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
FEATURES             Location/Qualifiers
     source          1..5575
                     /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"
     CDS             complement(4916..5575)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5490..5509
                     /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 tgatgagtta ataacaggcc tgctggtaat
     4681 cgcaggcctt tttatttttt tttttagcta gcagcagatt acgcgcagaa aaaaaggatc
     4741 tcaagaagat cctttgatct tattctacgg ggtctgacgc tcagtggaac gaaaactcac
     4801 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     4861 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacg
     4921 ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga
     4981 agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt
     5041 gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt
     5101 ttaaatcaaa actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa
     5161 taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata
     5221 tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag
     5281 tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt
     5341 ctttcattgc catacggaac tccggatgag cattcatcag gcgggcaaga atgtgaataa
     5401 aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca
     5461 gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt
     5521 tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc tccat
//
