LOCUS       sequence-348364-        4225 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lac inducible, AMP Resistant, non labelled gene mesh1 from D. 
            melanogaster on a low copy backbone. Used to induce controllable 
            hydrolysis of ppGpp in E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4225)
  AUTHORS   Buke F, Grilli J, Cosentino Lagomarsino M, Bokinsky G, Tans SJ
  TITLE     ppGpp is a bacterial cell size regulator.
  JOURNAL   Curr Biol. 2022 Feb 28;32(4):870-877.e5. doi: 
            10.1016/j.cub.2021.12.033. Epub 2022 Jan 5.
  PUBMED    34990598
REFERENCE   2  (bases 1 to 4225)
  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"; doi: 
            "10.1016/j.cub.2021.12.033"; journalName: "Curr Biol"; date: 
            "2022-02-28- 28"; volume: "32"; issue: "4"; pages: "870-877.e5"
COMMENT     Sequence Label: pMesh1
FEATURES             Location/Qualifiers
     source          1..4225
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     845..864
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(884..961)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1167..1186
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        join(1272..1277,1278..1295,1296..1302)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1310..1326
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     terminator      1928..1999
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2015..2042
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(2070..2087)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2204..2749)
                     /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(2284..2303)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2863..2957)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2987..3778,3779..3847))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3610..3629
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3848..3952)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(join(4026..4225,1..883))
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat
       61 cgtcgtatcc cactaccgag atgtccgcac caacgcgcag cccggactcg gtaatggcgc
      121 gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct
      181 cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt
      241 ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac
      301 gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga
      361 ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg
      421 gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag
      481 caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga
      541 gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca
      601 ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg
      661 cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt
      721 gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc
      781 gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga
      841 caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt
      901 gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt
      961 ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg
     1021 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac
     1081 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     1141 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     1201 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatcgtttag
     1261 gcaccccagg ctttacactt tatgcttccg gctcgtataa tgtgtggaat tgtgagcgga
     1321 taacaatttc agaattcaaa agatctagga ggaaaaaaat ggcaacatat cctagtgcaa
     1381 aatttatgga atgccttcag tacgccgcgt tcaaacatcg gcaacagcgc cggaaggacc
     1441 cgcaagagac cccgtacgtg aaccatgtta tcaacgtatc aaccatcctg tccgttgaag
     1501 cctgcattac tgacgaagga gtcctgatgg cggcgttatt gcacgacgtt gtagaggaca
     1561 ctgacgcttc cttcgaagat gttgaaaagt tgtttggtcc cgacgtgtgt ggcttggtgc
     1621 gcgaagtgac ggatgataag tccttggaga aacaggaacg caaacgctta cagattgaaa
     1681 acgcagccaa atcaagttgc cgtgctaaac tgatcaagct ggctgacaaa ctggacaatt
     1741 tacgggattt gcaggttaat acgccgacag gctggaccca ggagcgtcgg gatcagtatt
     1801 tcgtttgggc gaaaaaagtg gttgataatc tgcgtgggac taacgctaat cttgagctga
     1861 aattggatga gattttccgc caacgcggtc tgctgggatc ctaactcgag taaggatctc
     1921 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     1981 tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg ggcctttctg
     2041 cgtttatacc tagggatata ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt
     2101 cgactgcggc gagcggaaat ggcttacgaa cggggcggag atttcctgga agatgccagg
     2161 aagatactta acagggaagt gagagggccg cggcaaagcc gtttttccat aggctccgcc
     2221 cccctgacaa gcatcacgaa atctgacgct caaatcagtg gtggcgaaac ccgacaggac
     2281 tataaagata ccaggcgttt ccccctggcg gctccctcgt gcgctctcct gttcctgcct
     2341 ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc acgcctgaca
     2401 ctcagttccg ggtaggcagt tcgctccaag ctggactgta tgcacgaacc ccccgttcag
     2461 tccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccgga aagacatgca
     2521 aaagcaccac tggcagcagc cactggtaat tgatttagag gagttagtct tgaagtcatg
     2581 cgccggttaa ggctaaactg aaaggacaag ttttggtgac tgcgctcctc caagccagtt
     2641 acctcggttc aaagagttgg tagctcagag aaccttcgaa aaaccgccct gcaaggcggt
     2701 tttttcgttt tcagagcaag agattacgcg cagaccaaaa cgatctcaag aagatcatct
     2761 tattaatcag ataaaatatt tctagatttc agtgcaattt atctcttcaa atgtagcacc
     2821 tgaagtcagc cccatacgat ataagttgtt actagtgctt ggattctcac caataaaaaa
     2881 cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg
     2941 atctatcaac aggagtccaa gcgagctcgt aaacttggtc tgacagttac caatgcttaa
     3001 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3061 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3121 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3181 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     3241 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     3301 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     3361 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     3421 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     3481 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     3541 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     3601 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     3661 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     3721 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     3781 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     3841 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     3901 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     3961 cccgaaaagt gccacctgac gtcggtgcct aatgagtgag ctaacttaca ttaattgcgt
     4021 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4081 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc
     4141 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag
     4201 cggtccacgc tggtttgccc cagca
//
