LOCUS       sequence-203464-        2939 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control plasmid expressing GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2939)
  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 2939)
  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-GFP
FEATURES             Location/Qualifiers
     source          1..2939
                     /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             364..372
                     /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             384..413
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(462..490)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1085..1105
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1272..1289)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1406..1951)
                     /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(1486..1505)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2280..2939)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2854..2873
                     /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 acttttcgca ctagccgcta
      361 tattaaggag gtaataaatg gtggaacaaa agttgatcag cgaagaagac ttgggcagcg
      421 atggtagcgc gggtagtaaa ggagaagaac ttttcactgg agttgtccca attcttgttg
      481 aattagatgg tgatgttaat gggcacaaat tttctgtccg tggagagggt gaaggtgatg
      541 ctacaaacgg aaaactcacc cttaaattta tttgcactac tggaaaacta cctgttccgt
      601 ggccaacact tgtcactact ctgacctatg gtgttcaatg cttttcccgt tatccggatc
      661 acatgaaacg gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaacgca
      721 ctatatcttt caaagatgac gggacctaca agacgcgtgc tgaagtcaag tttgaaggtg
      781 atacccttgt taatcgtatc gagttaaagg gtattgattt taaagaagat ggaaacattc
      841 ttggacacaa actcgagtac aactttaact cacacaatgt atacatcacg gcagacaaac
      901 aaaagaatgg aatcaaagct aacttcaaaa ttcgccacaa cgttgaagat ggttccgttc
      961 aactagcaga ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag
     1021 acaaccatta cctgtcgaca caatctgtcc tttcgaaaga tcccaacgaa aagcgtgacc
     1081 acatggtcct tcttgagttt gtaactgctg ctgggattac acatggctaa taataatgat
     1141 gagttaataa cagcctgctg gtaatcgcag gcctttttat tttttttttt agatctagct
     1201 tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata
     1261 tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa
     1321 atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa
     1381 gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg
     1441 aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt
     1501 ttccccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca
     1561 ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca
     1621 gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta
     1681 tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca
     1741 gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac
     1801 tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt
     1861 ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca
     1921 agagattacg cgcagaccaa aacgatctca agaagatcat cttattaatc agataaaata
     1981 tttctagatt tcagtgcaat ttatctcttc aaatgtagca cctgaagtca gccccatacg
     2041 atataagttg tagcggccgc atctgaagta gctagcagca gattacgcgc agaaaaaaag
     2101 gatctcaaga agatcctttg atcttattct acggggtctg acgctcagtg gaacgaaaac
     2161 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     2221 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     2281 tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca
     2341 tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg
     2401 ccttgcgtat aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc
     2461 acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc
     2521 tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa
     2581 tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt
     2641 tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca
     2701 ccgtctttca ttgccatacg gaactccgga tgagcattca tcaggcgggc aagaatgtga
     2761 ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata
     2821 tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt
     2881 tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccat
//
