LOCUS       sequence-307550-        3411 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cloned with the codon-optimized mEGFP gene for expression in 
            Dictyostelium discoideum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3411)
  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 3411)
  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/DdmEGFP
FEATURES             Location/Qualifiers
     source          1..3411
                     /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(1015..1032)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1015..1031)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1024..1046)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1240..1259)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1359..1381
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1419..1437)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1505..1609
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1610..1678,1679..2470)
                     /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(1828..1847)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2641..3229
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3130..3149
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3383..3400
                     /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 gagatctatg gttagtaaag gagaggagct tttcactgga
      301 gtagtaccta ttttagtaga gttagatgga gatgtaaatg gacataaatt ctcagtttct
      361 ggtgaaggtg aaggagatgc tacatatgga aaattaactc ttaaattcat atgtacaact
      421 ggaaagttac ctgtaccctg gcctaccttg gttactactc ttacctacgg tgtacaatgt
      481 tttagtcgtt accctgatca tatgaaacaa cacgatttct tcaagtcagc catgccagaa
      541 ggatacgtac aagagcgtac catcttcttc aaagatgatg gtaattataa gactagagca
      601 gaggtaaaat ttgagggtga tacccttgta aatcgtatag agttaaaggg tatagatttc
      661 aaggaggatg gaaatatctt aggacacaag ttggaataca attacaattc tcataatgtt
      721 tacatcatgg ctgataagca aaagaatggt attaaagtta attttaaaat cagacacaat
      781 atagaggatg gttcagttca attagctgat cactaccaac aaaatacacc tattggagat
      841 ggtccagtat tgttaccaga taatcattat ttgagtactc aatctaaatt gagtaaagat
      901 cctaatgaaa aacgtgatca catggttctt ttggagtttg ttactgccgc aggaattacc
      961 ttgggtatgg atgaattgta taagtaggga tccccgggta ccgagctcga attcactggc
     1021 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     1081 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     1141 ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca
     1201 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc
     1261 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     1321 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     1381 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     1441 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     1501 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     1561 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     1621 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     1681 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     1741 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     1801 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     1861 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     1921 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     1981 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     2041 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     2101 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     2161 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     2221 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     2281 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     2341 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     2401 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     2461 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     2521 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     2581 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2641 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2701 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2761 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     2821 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2881 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2941 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3001 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3061 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3121 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3181 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3241 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     3301 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     3361 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag a
//
