LOCUS       sequence-203475-        4573 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of Cherry and GFP-PLC-delta-PH domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4573)
  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 4573)
  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: pClodAcytCh-PIP2PHGFP
FEATURES             Location/Qualifiers
     source          1..4573
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..69,70..861)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(219..238)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     RBS             1174..1196
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1727..1738
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             join(2979..2981,2982..2984,2985..3695)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2979..2981,2982..2984,2985..3695)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3024..3045)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3285..3304)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3632..3653
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3696..3761
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     complement(4383..4401)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4469..4573
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
       61 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
      121 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
      181 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
      241 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
      301 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
      361 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
      421 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
      481 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
      541 cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
      601 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
      661 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
      721 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
      781 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
      841 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
      901 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
      961 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccac catggacaca
     1021 gctaacacca cgtcgtccct atctgctgcc ctaggtctat gagtggttgc tggataactt
     1081 tacgggcatg cataaggctc gtataatata ttcagggaga ccacaacggt ttccctctac
     1141 aaataatttt gtttaacttt gagggacaat aattttgttt aactttaaga aggagatata
     1201 cattactagt atcgatggtc tcaaagggtg aagaggacaa catggcaata attaaagagt
     1261 tcatgagatt taaagtccat atggagggtt cagtcaacgg acacgagttc gagattgagg
     1321 gtgagggaga gggaaggcca tacgagggaa ctcagactgc aaagttgaag gtcacaaaag
     1381 gtggaccttt accattcgca tgggacatat tatcaccaca gtttatgtat ggttctaagg
     1441 catacgtcaa gcacccagct gatattccag actacttgaa gttgtctttt cctgaaggtt
     1501 tcaaatggga gagggtcatg aacttcgagg acggaggagt cgtcactgtc actcaggact
     1561 catctttgca ggatggagag tttatataca aagttaaatt gaggggaaca aacttcccat
     1621 cagacggacc agtcatgcaa aagaagacta tgggttggga ggcttcttca gagaggatgt
     1681 acccagagga cggagctttg aagggtgaga taaaacaaag gttgaaattg aaggacggtg
     1741 gtcactacga cgctgaagtt aagacaacat acaaggctaa gaagcctgtc caattgcctg
     1801 gtgcttataa cgtcaacatt aaattggaca ttacttctca caacgaggat tacactattg
     1861 tcgagcaata cgagagggct gagggaagac actctactgg aggtatggac gagttgtaca
     1921 aataataagt cgacgttaat aacaggcctg ctggtaatcg caggcctttt tatttttttt
     1981 tttaagatct agcgctgcgg acacatacaa agttgcccac agattccgtg gataagcagg
     2041 ggactaacat gtgaggcaaa acagcagggc cgcgccggtg gcgtttttcc ataggctccg
     2101 ccctcctgcc agagttcaca taaacagacg cttttccggt gcatctgtgg gagccgtgag
     2161 gctcaaccat gaatctgaca gtacgggcga aacccgacag gacttaaaga tccccaccgt
     2221 ttccggcggg tcgctccctc ttgcgctctc ctgttccgac cctgccgttt accggatacc
     2281 tgttccgcct ttctccctta cgggaagtgt ggcgctttct catagctcac acactggtat
     2341 ctcggctcgg tgtaggtcgt tcgctccaag ctgggctgta agcaagaact ccccgttcag
     2401 cccgactgct gcgccttatc cggtaactgt tcacttgagt ccaacccgga aaagcacggt
     2461 aaaacgccac tggcagcagc cattggtaac tgggagttcg cagaggattt gtttagctaa
     2521 acacgcggtt gctcttgaag tgtgcgccaa agtccggcta cactggaagg acagatttgg
     2581 ttgctgtgct ctgcgaaagc cagttaccac ggttaagcag ttccccaact gacttaacct
     2641 tcgatcaaac cacctcccca ggtggttttt tcgaaggtta agtcagttgg ggaactgctt
     2701 aaccttcgat caaaccacct ccccaggtgg ttttttcgtt tacagggcaa aagattacgc
     2761 gcagaaaaaa aggatctcaa gaagatcctt tgatcagcgg ccgccacagc taacaccacg
     2821 tcgtccctat ctgctgccct aggtctatga gtggttgctg gataacttta cgggcatgca
     2881 taaggctcgt ataatatatt cagggagacc acaacggttt ccctctacaa ataattttgt
     2941 ttaactttca attgaaggag atatacatac tagtatcgat ggtgagcaag ggcgaggagc
     3001 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     3061 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     3121 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     3181 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     3241 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     3301 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     3361 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     3421 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     3481 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     3541 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     3601 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     3661 ccgggatcac tctcggcatg gacgagctgt acaagtccgg actcagatct cgagctcaag
     3721 cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc catggactcg ggccgggact
     3781 tcctgaccct gcacggccta caggatgatg aggatctaca ggcgctgctg aagggcagcc
     3841 agctcctgaa ggtgaagtcc agctcatgga ggagagagcg cttctacaag ttgcaggagg
     3901 actgcaagac catctggcag gagtcccgca aggtcatgcg gaccccggag tcccagctgt
     3961 tctccatcga ggacattcag gaggtgcgaa tggggcaccg cacggagggt ctggagaagt
     4021 tcgcccgtga tgtgcccgag gaccgctgct tctccattgt cttcaaggac cagcgcaata
     4081 cactagacct catcgcccca tcgccagctg atgcccagca ctgggtgctg gggctgcaca
     4141 agatcatcca ccactcaggc tccatggacc agcgtcagaa gctacagcac tggattcact
     4201 cctgcttgcg aaaagctgac aaaaacaagg acaacaagat gagcttcaag gagctgcaga
     4261 acttcctgaa ggagctcaac atccagtaat gataaaccgg ttgatgagtt aataacaggc
     4321 ctgctggtaa tcgcaggcct ttttattttt ttttttgcta gcaaattctt gaagacgaaa
     4381 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     4441 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4501 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4561 aaaaaggaag agt
//
