LOCUS       sequence-307479-        3446 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for conony screening without an inducer (e.g. 
            IPTG); TED module (Plac-lacO-mlcR25-mEGFP).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3446)
  AUTHORS   Kondo T, Yumura S
  TITLE     An improved molecular tool for screening bacterial colonies using 
            GFP expression enhanced by a Dictyostelium sequence.
  JOURNAL   Biotechniques. 2020 Feb;68(2):91-95. doi: 10.2144/btn-2019-0127. 
            Epub 2019 Dec 11.
  PUBMED    31825246
REFERENCE   2  (bases 1 to 3446)
  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: 
            "10.2144/btn-2019-0127"; journalName: "Biotechniques"; date: 
            "2020-02"; volume: "68"; issue: "2"; pages: "91-95"
COMMENT     Sequence Label: pUC19/mlcR25-mEGFP
FEATURES             Location/Qualifiers
     source          1..3446
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(142..147,148..165,166..172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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)."
     primer_bind     185..207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     204..220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1050..1067)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1050..1066)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1059..1081)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1275..1294)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1394..1416
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1454..1472)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1540..1644
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1645..1713,1714..2505)
                     /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(1863..1882)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2676..3264
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3165..3184
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3418..3435
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
      241 atgcctgcag gtcgactcta gacgttaata ctctcttcag taaaaaagga ggtggtggat
      301 ctatggttag taaaggagag gagcttttca ctggagtagt acctatttta gtagagttag
      361 atggagatgt aaatggacat aaattctcag tttctggtga aggtgaagga gatgctacat
      421 atggaaaatt aactcttaaa ttcatatgta caactggaaa gttacctgta ccctggccta
      481 ccttggttac tactcttacc tacggtgtac aatgttttag tcgttaccct gatcatatga
      541 aacaacacga tttcttcaag tcagccatgc cagaaggata cgtacaagag cgtaccatct
      601 tcttcaaaga tgatggtaat tataagacta gagcagaggt aaaatttgag ggtgataccc
      661 ttgtaaatcg tatagagtta aagggtatag atttcaagga ggatggaaat atcttaggac
      721 acaagttgga atacaattac aattctcata atgtttacat catggctgat aagcaaaaga
      781 atggtattaa agttaatttt aaaatcagac acaatataga ggatggttca gttcaattag
      841 ctgatcacta ccaacaaaat acacctattg gagatggtcc agtattgtta ccagataatc
      901 attatttgag tactcaatct aaattgagta aagatcctaa tgaaaaacgt gatcacatgg
      961 ttcttttgga gtttgttact gccgcaggaa ttaccttggg tatggatgaa ttgtataagt
     1021 agggatcccc gggtaccgag ctcgaattca ctggccgtcg ttttacaacg tcgtgactgg
     1081 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     1141 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     1201 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     1261 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     1321 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     1381 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     1441 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     1501 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     1561 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     1621 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     1681 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     1741 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     1801 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     1861 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     1921 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     1981 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     2041 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     2101 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     2161 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     2221 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     2281 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     2341 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     2401 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     2461 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     2521 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     2581 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     2641 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     2701 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     2761 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     2821 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     2881 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     2941 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     3001 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     3061 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     3121 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     3181 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     3241 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     3301 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     3361 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     3421 gagtcagtga gcgaggaagc ggaaga
//
