LOCUS       sequence-307549-        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    105..126
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(141..146,147..164,165..171)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    179..195
                     /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     184..206
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     203..219
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     203..219
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1009..1026)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1009..1025)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1018..1040)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1234..1253)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1353..1375
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1413..1431)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1499..1603
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1604..1672,1673..2464)
                     /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(1822..1841)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2635..3223
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3124..3143
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3377..3394
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
       61 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
      121 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
      181 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttgca
      241 tgcctgcagg tcgactctag aagatctatg gtaagtaagg gagaggaatt aataaaagaa
      301 aatatgcgta tgaaggttgt tatggaagga tcagtaaatg gacaccaatt caagtgtact
      361 ggtgagggag agggtagacc atacgaggga gtacaaacca tgagaataaa ggttatcgaa
      421 ggtggaccat tgccttttgc cttcgatatt ttagctacaa gtttcatgta tggttctcgt
      481 acatttatta aatatccagc agatatacca gattttttca aacaatcttt tcctgaggga
      541 tttacttggg agagagtaac cagatatgaa gatggtggag ttgttacagt tacacaagat
      601 accagtttgg aagatggaga gttagtttac aatgtaaaag taagaggagt aaatttccca
      661 agtaatggtc cagtaatgca aaagaagaca aagggttggg aaccaaatac cgaaatgatg
      721 tatcctgctg atggaggttt aagaggatat acagatatag cattgaaggt tgatggtggt
      781 ggacatttac attgtaattt cgtaaccacc taccgttcaa aaaagaccgt aggtaatata
      841 aagatgccag gagtacatgc tgtagatcat agattggaga gaatagaaga gtcagataat
      901 gagacttatg tagtacaaag agaagttgcc gtagccaaat acagtaatct tggaggtggt
      961 atggacgagc tgtacaagta aggatccccg ggtaccgagc tcgaattcac tggccgtcgt
     1021 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     1081 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     1141 gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg
     1201 cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     1261 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     1321 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     1381 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     1441 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     1501 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     1561 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     1621 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     1681 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     1741 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     1801 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     1861 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     1921 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     1981 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     2041 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     2101 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     2161 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     2221 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     2281 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     2341 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     2401 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     2461 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     2521 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     2581 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     2641 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     2701 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     2761 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     2821 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     2881 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     2941 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3001 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3061 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3121 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3181 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3241 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     3301 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     3361 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagag
//
