LOCUS       sequence-298585-        3406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cloned with the codon-optimized mRuby3 gene for expression in 
            Dictyostelium discoideum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3406)
  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 3406)
  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/DdmRuby3
FEATURES             Location/Qualifiers
     source          1..3406
                     /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(1010..1027)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1010..1026)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1019..1041)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1235..1254)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1354..1376
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1414..1432)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1500..1604
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1605..1673,1674..2465)
                     /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(1823..1842)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2636..3224
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3125..3144
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3378..3395
                     /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 gaagatctat ggtaagtaag ggagaggaat taataaaaga
      301 aaatatgcgt atgaaggttg ttatggaagg atcagtaaat ggacaccaat tcaagtgtac
      361 tggtgaggga gagggtagac catacgaggg agtacaaacc atgagaataa aggttatcga
      421 aggtggacca ttgccttttg ccttcgatat tttagctaca agtttcatgt atggttctcg
      481 tacatttatt aaatatccag cagatatacc agattttttc aaacaatctt ttcctgaggg
      541 atttacttgg gagagagtaa ccagatatga agatggtgga gttgttacag ttacacaaga
      601 taccagtttg gaagatggag agttagttta caatgtaaaa gtaagaggag taaatttccc
      661 aagtaatggt ccagtaatgc aaaagaagac aaagggttgg gaaccaaata ccgaaatgat
      721 gtatcctgct gatggaggtt taagaggata tacagatata gcattgaagg ttgatggtgg
      781 tggacattta cattgtaatt tcgtaaccac ctaccgttca aaaaagaccg taggtaatat
      841 aaagatgcca ggagtacatg ctgtagatca tagattggag agaatagaag agtcagataa
      901 tgagacttat gtagtacaaa gagaagttgc cgtagccaaa tacagtaatc ttggaggtgg
      961 tatggacgag ctgtacaagt aaggatcccc gggtaccgag ctcgaattca ctggccgtcg
     1021 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     1081 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     1141 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt
     1201 gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     1261 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     1321 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     1381 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     1441 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     1501 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     1561 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     1621 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     1681 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     1741 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     1801 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     1861 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     1921 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     1981 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     2041 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     2101 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     2161 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     2221 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     2281 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     2341 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     2401 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     2461 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     2521 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     2581 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     2641 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     2701 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     2761 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     2821 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     2881 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     2941 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3001 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3061 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3121 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3181 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3241 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3301 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     3361 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaaga
//
