LOCUS       sequence-348365-        4948 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lac inducible, AMP Resistant, fluor labelled (CFP) 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 4948)
  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 4948)
  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..4948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1035..1051
                     /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)."
     promoter        complement(join(1059..1065,1066..1083,1084..1089))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     primer_bind     complement(1175..1194)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1400..1477
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1478..2560
                     /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"
     primer_bind     complement(1497..1516)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        2634..2738
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2739..2807,2808..3599)
                     /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     complement(2957..2976)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3629..3723
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      3837..4382
                     /direction=RIGHT
                     /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     4283..4302
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4499..4516
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4544..4571)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(4587..4658)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggtgatatac acattatcgc taatcgcatt atattccagt ttatggccca ggatattacc
       61 atcttcctta aaatcgatgc ccttcagctc aatacgattc accagagtgt cgccttcaaa
      121 cttcacctct gcgcgagtct tgtaattacc atcatcttta aagaaaatgg tgcgctcctg
      181 aacgtaaccc tctggcatgg cgctcttaaa aaaatcgtgc tgcttcatgt ggtctggata
      241 gcgagcgaag cattgaacac cccaggtcag ggtggtaacc agggtaggcc aaggcaccgg
      301 cagcttaccc gtcgtacaaa tgaacttcag ggtcagctta ccgtaggttg cgtcgccttc
      361 accctcgccg ctaacgctga acttatgacc gttgacatca ccgtccagtt cgaccaggat
      421 aggaaccaca ccagtaaaca gctcctcgcc cttgctcata gatcccagca gaccgcgttg
      481 gcggaaaatc tcatccaatt tcagctcaag attagcgtta gtcccacgca gattatcaac
      541 cacttttttc gcccaaacga aatactgatc ccgacgctcc tgggtccagc ctgtcggcgt
      601 attaacctgc aaatcccgta aattgtccag tttgtcagcc agcttgatca gtttagcacg
      661 gcaacttgat ttggctgcgt tttcaatctg taagcgtttg cgttcctgtt tctccaagga
      721 cttatcatcc gtcacttcgc gcaccaagcc acacacgtcg ggaccaaaca acttttcaac
      781 atcttcgaag gaagcgtcag tgtcctctac aacgtcgtgc aataacgccg ccatcaggac
      841 tccttcgtca gtaatgcagg cttcaacgga caggatggtt gatacgttga taacatggtt
      901 cacgtacggg gtctcttgcg ggtccttccg gcgctgttgc cgatgtttga acgcggcgta
      961 ctgaaggcat tccataaatt ttgcactagg atatgttgcc atttttttcc tcctagatct
     1021 tttgaattct gaaattgtta tccgctcaca attccacaca ttatacgagc cggaagcata
     1081 aagtgtaaag cctggggtgc ctaaacgatc tcgatcctct acgccggacg catcgtggcc
     1141 ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat caccgatggg
     1201 gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg tatggtggca
     1261 ggccccgtgg ccgggggact gttgggcgcc atctccttgc atgcaccatt ccttgcggcg
     1321 gcggtgctca acggcctcaa cctactactg ggctgcttcc taatgcagga gtcgcataag
     1381 ggagagcgtc gagatcccgg acaccatcga atggcgcaaa acctttcgcg gtatggcatg
     1441 atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga
     1501 tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag
     1561 ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat
     1621 tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac
     1681 ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga
     1741 tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa
     1801 agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct
     1861 ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct
     1921 tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg
     1981 actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc
     2041 attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa
     2101 tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca
     2161 aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca
     2221 gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggacat
     2281 ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac
     2341 catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc
     2401 tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac
     2461 caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     2521 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtaa
     2581 gttagctcac tcattaggca ccgacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     2641 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2701 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     2761 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     2821 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     2881 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     2941 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     3001 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     3061 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     3121 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     3181 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     3241 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     3301 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     3361 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     3421 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     3481 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     3541 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     3601 tgtcagacca agtttacgag ctcgcttgga ctcctgttga tagatccagt aatgacctca
     3661 gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag
     3721 aatccaagca ctagtaacaa cttatatcgt atggggctga cttcaggtgc tacatttgaa
     3781 gagataaatt gcactgaaat ctagaaatat tttatctgat taataagatg atcttcttga
     3841 gatcgttttg gtctgcgcgt aatctcttgc tctgaaaacg aaaaaaccgc cttgcagggc
     3901 ggtttttcga aggttctctg agctaccaac tctttgaacc gaggtaactg gcttggagga
     3961 gcgcagtcac caaaacttgt cctttcagtt tagccttaac cggcgcatga cttcaagact
     4021 aactcctcta aatcaattac cagtggctgc tgccagtggt gcttttgcat gtctttccgg
     4081 gttggactca agacgatagt taccggataa ggcgcagcgg tcggactgaa cggggggttc
     4141 gtgcatacag tccagcttgg agcgaactgc ctacccggaa ctgagtgtca ggcgtggaat
     4201 gagacaaacg cggccataac agcggaatga caccggtaaa ccgaaaggca ggaacaggag
     4261 agcgcacgag ggagccgcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4321 gccaccactg atttgagcgt cagatttcgt gatgcttgtc aggggggcgg agcctatgga
     4381 aaaacggctt tgccgcggcc ctctcacttc cctgttaagt atcttcctgg catcttccag
     4441 gaaatctccg ccccgttcgt aagccatttc cgctcgccgc agtcgaacga ccgagcgtag
     4501 cgagtcagtg agcgaggaag cggaatatat ccctaggtat aaacgcagaa aggcccaccc
     4561 gaaggtgagc cagtgtgact ctagtagaga gcgttcaccg acaaacaaca gataaaacga
     4621 aaggcccagt ctttcgactg agcctttcgt tttatttgat gcctggagat ccttactcga
     4681 gttaggatcc ttatttatac agctcatcca tgccatgcgt gatgccagca gccgttacaa
     4741 attccagcag caccatgtga tcgcgcttct cattaggatc tttgctcagc ttagattgag
     4801 tgctcagata gtgattgtcc ggcagcagaa caggaccgtc gccaatcggg gtattttgtt
     4861 gataatggtc agccagttgc acagagccgt cctcgatatt atggcgaatt ttaaaatgcg
     4921 ccttaatacc attcttttgt ttatcggc
//
