LOCUS       sequence-298589-        8454 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A expression vector in Dictyostelium discoideum containing the 
            Blasticidin S resistant gene and the TED module for GFP-negative 
            screening.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8454)
  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 8454)
  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: pDYU1B-TED
FEATURES             Location/Qualifiers
     source          1..8454
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    699..720
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(735..740,741..758,759..765)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    773..789
                     /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     778..800
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     797..813
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5912..5929)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5912..5928)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5921..5943)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6137..6156)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6256..6278
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6316..6334)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6402..6506
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6507..6575,6576..7367)
                     /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(6725..6744)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7538..8126
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8027..8046
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    8414..8435
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 caggaaacag ctatgaccat gattacgcca agctcggcgc gccattggga tggaacgggc
       61 ccctagataa aaaaaatttt tatttatttt tatttatttt gaattaaata gattacaaat
      121 taattaatcc catcaaatct ttaaaaaaaa aaatggttta aaaaaacttg ggttggttaa
      181 ttattatttg aaaattttaa aacccaaatt aaaaaaaaaa aatgggattc aaaaattttt
      241 tttttttttt tttttttttt tttttttttt tttttttttt tttttttttt tttcagattg
      301 cataaaaaga tttttttttt tttttttttc ttatttctta aaacaaataa attaaattaa
      361 ataaaaaata aaatctagac cagatctccc tcgagccggt accccggatc caacgccagc
      421 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      481 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
      541 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
      601 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
      661 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
      721 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
      781 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct tgcatgcctg
      841 caggtcgact ctagacgtta atactctctt cagtaaaaaa ggaggtggtg gatctatggt
      901 tagtaaagga gaggagcttt tcactggagt agtacctatt ttagtagagt tagatggaga
      961 tgtaaatgga cataaattct cagtttctgg tgaaggtgaa ggagatgcta catatggaaa
     1021 attaactctt aaattcatat gtacaactgg aaagttacct gtaccctggc ctaccttggt
     1081 tactactctt acctacggtg tacaatgttt tagtcgttac cctgatcata tgaaacaaca
     1141 cgatttcttc aagtcagcca tgccagaagg atacgtacaa gagcgtacca tcttcttcaa
     1201 agatgatggt aattataaga ctagagcaga ggtaaaattt gagggtgata cccttgtaaa
     1261 tcgtatagag ttaaagggta tagatttcaa ggaggatgga aatatcttag gacacaagtt
     1321 ggaatacaat tacaattctc ataatgttta catcatggct gataagcaaa agaatggtat
     1381 taaagttaat tttaaaatca gacacaatat agaggatggt tcagttcaat tagctgatca
     1441 ctaccaacaa aatacaccta ttggagatgg tccagtattg ttaccagata atcattattt
     1501 gagtactcaa tctaaattga gtaaagatcc taatgaaaaa cgtgatcaca tggttctttt
     1561 ggagtttgtt actgccgcag gaattacctt gggtatggat gaattgtata agtagggatc
     1621 cccgagctca tcatgatgaa agtgcttcac ataaaaataa taataataat ataacaataa
     1681 taatatttaa atgtataata aaatttaatt actttttttt taatggttgt tgatctttat
     1741 ccgaccttaa aaaaaaaaaa ataaaaccaa tagctattgt tttttttttt aattgttttt
     1801 ttatttttta ttattacttt aattatcatt ttttaaatta caaaaaaaat taaaaatcca
     1861 gatattaagg tatttgcact attgctttaa cgttaagctt gatggctcaa ataacggtat
     1921 ggaaaagtca gtaactcaag aaaaaaacat ttcaaataat ttaaaaatta atatatattt
     1981 aattttcttt ttaattattt ttttaattaa ataaagtttt attatttttt aagagtaatt
     2041 attgctcttt tttcatttga aacaccagaa gctaaacgta attgttgttg actgaaattt
     2101 tttatttttt ttggggtaat aggatttcct tttttatgaa gattaatatc tttgactcgt
     2161 gaaacattct ttttaacttt tgttttttct gttggtttat catttgtttt ttcactaatt
     2221 tcaataccat cttgacgttc attcataact tcatcttttt tttttcctgt ttctgtatct
     2281 tcttctattt ttttttcttt atctttttct ttatcttctt cttgttcttc ctcttctttt
     2341 tcttcttctg atactgcagg tgtttcttct tcttcttctt ccgatattgt cggtttttct
     2401 acttcttctt cttgttcttc ttcttcttct tcttcttctt cttcttcttc ttcttcttct
     2461 tcttcttccg gtaatttatt aattatattt ctttttttat atgaattacg tttggtttgt
     2521 gcagtaattt ccttacatag agtgcagctt tcaagaaaaa tttcaatttc ttcgtttgtt
     2581 gcataataac cactgtcttt gatatgatta aacatttttg attttcttaa atgctttcct
     2641 tctttaatat gaaaattatc gaattctaat tcattaagaa caataagctc ccctaattta
     2701 aaaaattagt taaaataaat taaaatgaac atgtataaag atggatttta ccattttttg
     2761 aaattctaaa taacttttct tcatctccaa tctttttgac tgaaaaacga tttttaattg
     2821 aagttattgt tctgtgagtg ttttgaatcg cccatttctc taaatcagtt tgagatagtg
     2881 ttttataatc tgaattgtta tacacaactt ttgctctatt aaccaaatat ttaaagattt
     2941 catcatcaac tgaatatttt gactttacga ttcttgtcca aaaaacaatt tctactacta
     3001 tcatttttta tttataaaat aatttaaata caaaaatgaa tttttttttt tttaaaaaaa
     3061 aaaaaatttg aaaaaaaaaa aaaaaaaatt ttaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     3121 aaaaaatcaa ataaaaagta aaaaataaaa accgaaaaac attcattgta atttcaaatg
     3181 tcaacaacaa caacaacaac aaacaaaagg aaaattgctc ctccatcgaa caacacacca
     3241 aaaaaacaaa atgttgaaat tttaggagtt caaaacattc acaattcata ttcattagct
     3301 ggagttttaa ttggcgcaag cgaagaattt aaaaacaaat tggggaaaat ttttcgtctt
     3361 gatttagcta acagctctac ttataacctt gtttcacaag ttcaatatca cgttgttaat
     3421 gctctcaaat ttgttaaaaa gcctgtccct gttgatggtg aactatttaa aacaaatcaa
     3481 gttcaacagg taagcgatca acttgaaggt gtcctttcaa acctcaccat gatgatttca
     3541 aatgcagata aaaaaaaaaa agaatcatcc cagtatccca atctattaaa atcaatttat
     3601 gttaaagttt atactgaagc tgtaacagaa aggacatcct caaatgaaat ttctgaagcc
     3661 aaccttttga aatctttaaa tcaatacatt tccaatacca taacccaaat ctatatcgac
     3721 tcttctgatc caccatttcc aaatattgat tcacttaaag agtttccata tttttgttca
     3781 ctatcagctg gtaaaatttc actccaacgg tttattgata attactcata tctcattgat
     3841 aataattttt taaaaggtat ctttattcaa atttctcacc aaacttcgtt tgttggtgat
     3901 gtttttggtt atattaattc tgacatcaag cttgacgata ttatgctcga tttttcttcg
     3961 agatccccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgctt
     4021 tgcctgtttc cggcaccaga agcggtgccg gaaagctggc tggagtgcga tcttcctgag
     4081 ggtccatcaa ttgttcacag aaaatgtttt taaattattt aataaataat aaaaaaacaa
     4141 attgttgtaa taatctaata ttttcttttt tttttaattt ttttttttta aatcttaata
     4201 attattaagg tattttaatt tttttttttt tttttttttt tttttttttt tttttctatc
     4261 aaaaaaatca aatatattta aaaaatttat tatttacaga tacattttga atggtgaaga
     4321 taaatatatg cattagatgt aaaacagcca aagagtatga aaatcaaaaa gataaagctg
     4381 atccgggacc aaagcggcca tcgtgcctcc ccactcctgc agttcggggg catggatgcg
     4441 cggatagccg ctgctggttt cctggatgcc gacggatttg cactgccggt agaactccgc
     4501 gaggtcgtcc agcctcaggc agcagctgaa ccaactcgcg aggggatcga gcccggggtg
     4561 ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa
     4621 aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg
     4681 caggttgggc gtcgcttggt cggtcatttc gaaccccaga gtcccgctta accttcccaa
     4741 acgtaaccag aaggaagtaa ttcacggatt cctactgctg ttggttgtcc atcagaatct
     4801 ttaacaatgg ctttaatacc aggatgaaga tccaacaaaa cttgtctaca acgaccacat
     4861 ggagataaaa taccacgatt ctcatttcca attgctacta tacaagtaag atttccggct
     4921 gctgctgcgg ctgcggttcc taatacaaca agctcagcac aaggacctcc tgtgaagtga
     4981 tatacattta ctccggtaaa aattctacca tcagaactaa gagcggcaga ggcaactgag
     5041 taatcttcac ttatagggat agaatttatg gtagcagttg cacgctcaat taaagtagac
     5101 tcctcttgag acaatggctt tgccatgcga aacgatccag cttgaacatc ttcaccatcc
     5161 attttatatt atatttattt atttgattat tttttgaatt aattaaaaaa aaaaaaaaaa
     5221 tttcatttta taatctcaga aacctcaaaa aaaaaaaaaa taaaaaataa aaaatataaa
     5281 aaaataaaaa taaaatccca attttaaagc gaaaaaccac ccatggtttg aaaatttcaa
     5341 tcaatttcaa ataactttac ttaaaaaaaa cccatttttt atttaaaaat ttctttttta
     5401 attaggcatc agaatatcaa ttctttgttt attacttgaa ttatttagga aaaaaaaaaa
     5461 aaaaaaaatc atccatttgt acaaaaactt taaaattcta tttttttttt tttttttata
     5521 attattattt tttcatatct cttttttttt tatataaaat atccatcatt attcttcaaa
     5581 ataaaaaaat aattacaaaa agtgaataat aattaaataa ttaataaccc tttacacttt
     5641 tttttttctt tttacagttt ttatatttat tttaatttta ttattttgcg caactttgaa
     5701 aaaataaaaa agataaagtt tttataaata tctatcaaaa agatatcgtt gttgattgat
     5761 tattttggat aatttttctt taaattgggg caaatctgta attttcagat taataagatt
     5821 tttccttttt atttttcttt ttagtttttg taatttttaa aaataaacct ctgaattcca
     5881 actagtggaa catcccaata cgcgtcaatt cactggccgt cgttttacaa cgtcgtgact
     5941 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     6001 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     6061 gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     6121 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     6181 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     6241 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     6301 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     6361 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     6421 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     6481 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     6541 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     6601 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     6661 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     6721 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     6781 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     6841 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     6901 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     6961 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     7021 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     7081 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     7141 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     7201 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     7261 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     7321 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     7381 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     7441 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     7501 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     7561 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     7621 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     7681 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     7741 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     7801 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     7861 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     7921 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     7981 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     8041 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     8101 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     8161 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     8221 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     8281 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     8341 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     8401 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct caca
//
