LOCUS       sequence-244258-        3782 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Slot A, contains mKO2-MarA (step 1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3782)
  AUTHORS   Santos-Moreno J, Schaerli Y
  TITLE     A Framework for the Modular and Combinatorial Assembly of Synthetic 
            Gene Circuits.
  JOURNAL   ACS Synth Biol. 2019 Jul 19;8(7):1691-1697. doi: 
            10.1021/acssynbio.9b00174. Epub 2019 Jun 24.
  PUBMED    31185158
REFERENCE   2  (bases 1 to 3782)
  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: 
            "10.1021/acssynbio.9b00174"; journalName: "ACS Synth Biol"; date: 
            "2019-07-19- 19"; volume: "8"; issue: "7"; pages: "1691-1697"
COMMENT     Sequence Label: pTU-A-010
FEATURES             Location/Qualifiers
     source          1..3782
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    289..308
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     RBS             313..335
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             343..996
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     primer_bind     complement(1709..1727)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1723..1770
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1779..1865
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1957..1984
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(2113..2904)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDDGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(2905..2996)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3044..3782)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
ORIGIN
        1 ttttctactg aaccgctcta gatttcagtg caatttatct cttcaaatgt agcacctgaa
       61 gtcagcccca tacgatataa gttgtaattc tcatgttagt catgccccgc gcccaccgga
      121 aggagctgac tgggttgctc ctaggatgca gtgactcgca catttccccg aaaagtgcca
      181 cctgaattca taggatagat tctggaaact ttaccgtccg agctcaggct taccttactc
      241 gagcaataaa cagttgatag ggcttctccg ttacagcctg cggtccgggt tcactgccgt
      301 ataggcagta attttgttta actttaagaa ggagatatac atatggtttc ggttatcaaa
      361 ccagagatga aaatgcgtta ctatatggat ggttcagtaa atggtcacga atttactatt
      421 gagggcgagg gtacgggacg cccatacgag gggcaccagg aaatgacttt acgcgtcaca
      481 atggctgaag gcgggcctat gccgtttgcg ttcgatcttg ttagtcatgt cttttgttac
      541 ggtcaccgtg tatttactaa ataccccgag gaaattccag actatttcaa acaagccttc
      601 ccggaaggtt tgtcttggga gcgcagttta gagtttgaag acggtggctc ggccagcgtg
      661 tcagctcata ttagtcttcg cggcaataca ttttatcaca agtcaaagtt caccggcgtg
      721 aacttccccg cagacggccc aatcatgcag aatcaaagtg ttgattggga accgtccaca
      781 gagaagatta cagcttccga tggagtctta aagggcgatg taaccatgta cttaaaatta
      841 gaagggggag ggaaccataa atgtcagatg aagactacct ataaggccgc aaaagagatt
      901 cttgaaatgc ccggagacca ctacattggg catcgtttgg tccgtaagac agaaggaaat
      961 attactgaac aggtcgaaga cgctgtggca cacagcatgt cccgccgtaa tactgacgcc
     1021 atcacaatcc acagcatcct ggattggatt gaagacaact tggagtcgcc gttgagttta
     1081 gaaaaagtta gtgaacgtag tggttactca aagtggcacc ttcagcgcat gtttaagaag
     1141 gaaacgggtc attcattggg tcaatatatt cgttctcgca agatgactga aattgcccag
     1201 aaattgaaag agtctaatga acctattttg tacctggcgg agcgttacgg ctttgaaagt
     1261 cagcaaaccc ttacacgtac cttcaagaat tactttgacg ttccaccaca caaatatcgt
     1321 atgaccaaca tgcagggtga gtcacgtttt ttgcatccgt tgaatcatta caattcctaa
     1381 taatcgctgg gacgcccgcc atggttcagc caaaaaactt aagaccgccg gtcttgtcca
     1441 ctaccttgca gtaatgcggt ggacaggatc ggcggttttc ttttctcttc tcaattcttc
     1501 tgacctgtaa cgaataatag atagtaaagt agtctccgat tgagttttct ctgccgagtc
     1561 ccacccagtt ctgtgatttc agtaagttgg taattgatac actgttgcga gaactgctgc
     1621 ctggtagtag ataggttgtt attgagtaag aaggtaaagt gaacgaaatc cctgaaactg
     1681 agactgtaga aaataagctt tgctgccacc gctgagcaat aactagcata accccttggg
     1741 gcctctaaac gggtcttgag gggttttttg ccaggcatca aataaaacga aaggctcagt
     1801 cggaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     1861 caaatccgcc gggagcggat ttgaacgttg tgaagcaacg gcccggaggg tggcgggcag
     1921 gacgcccgcc ataaactgcc aggcatcaaa ctaagcagaa ggccatcctg acggatggcc
     1981 tttttgcgtt tcagatctac cggtaaacca gcaatagaca taagcggcta tttaacgacc
     2041 ctgccctgaa ccgacgaccg ggtcatcgtg gccggatctt gcggcccctc ggcttgaacg
     2101 aattgttaga cattatttgc cgactacctt ggtgatctcg cctttcacgt agtggacaaa
     2161 ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct tcttgtccaa gataagcctg
     2221 tctagcttca agtatgacgg gctgatactg ggccggcagg cgctccattg cccagtcggc
     2281 agcgacatcc ttcggcgcga ttttgccggt tactgcgctg taccaaatgc gggacaacgt
     2341 aagcactaca tttcgctcat cgccagccca gtcgggcggc gagttccata gcgttaaggt
     2401 ttcatttagc gcctcaaata gatcctgttc aggaaccgga tcaaagagtt cctccgccgc
     2461 tggacctacc aaggcaacgc tatgttctct tgcttttgtc agcaagatag ccagatcaat
     2521 gtcgatcgtg gctggctcga agatacctgc aagaatgtca ttgcgctgcc attctccaaa
     2581 ttgcagttcg cgcttagctg gataacgcca cggaatgatg tcgtcgtgca caacaatggt
     2641 gacttctaca gcgcggagaa tctcgctctc tccaggggaa gccgaagttt ccaaaaggtc
     2701 gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa ccagcaaatc
     2761 aatatcactg tgtggcttca ggccgtcatc cactgcggag ccgtacaaat gtacggccag
     2821 caacgtcggt tcgagatggc gctcgatgac gccaactacc tctgatagtt gagtcgatac
     2881 ttcggcgatc accgcttccc tcatactctt cctttttcaa tattattgaa gcatttatca
     2941 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagc
     3001 tagctcactc ggtcgctacg ctccgggcgt gagactgcgg cgggcgctgc ggacacatac
     3061 aaagttaccc acagattccg tggataagca ggggactaac atgtgaggca aaacagcagg
     3121 gccgcgccgg tggcgttttt ccataggctc cgccctcctg ccagagttca cataaacaga
     3181 cgcttttccg gtgcatctgt gggagccgtg aggctcaacc atgaatctga cagtacgggc
     3241 gaaacccgac aggacttaaa gatccccacc gtttccggcg ggtcgctccc tcttgcgctc
     3301 tcctgttccg accctgccgt ttaccggata cctgttccgc ctttctccct tacgggaagt
     3361 gtggcgcttt ctcatagctc acacactggt atctcggctc ggtgtaggtc gtttgctcca
     3421 agctgggctg taagcaagaa ctccccgttc agcccgactg ctgcgcctta tccggtaact
     3481 gttcacttga gtccaacccg gaaaagcacg gtaaaacgcc actggcagca gccattggta
     3541 actgggagtt cgcagaggat ttgtttagct aaacacgcgg ttgctcttga agtgtgcgcc
     3601 aaagtccggc tacactggaa ggacagattt ggttgctgtg ctctgcgaaa gccagttacc
     3661 acggttaagc agttccccaa ctgacttaac cttcgatcaa accacctccc caggtggttt
     3721 tttcgtttac agggcaaaag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3781 tc
//
