LOCUS       sequence-244535-        3516 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Slot C, contains RepA70-mKate2 (step 1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3516)
  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 3516)
  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-C-010
FEATURES             Location/Qualifiers
     source          1..3516
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    287..306
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     RBS             311..333
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1443..1461)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1457..1504
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1513..1599
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1691..1718
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(1847..2638)
                     /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(2639..2730)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2778..3516)
                     /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 gaagtattgg taatcgttga aaactcagtc gacgcactta ctgaagacgt
      241 cctattacac tcgtcgttgg aaactgaaga tcagcctgcg gtccgggttc actgccgtat
      301 aggcagtaat tttgtttaac tttaagaagg agatatacat atgaatcagt cattcatctc
      361 ggacatctta tatgccgaca tcgaatcgaa ggctaaggaa cttacagtca attccaacaa
      421 tactgtccag ccggtcgcgc ttatgcgctt aggagttttc gttcccaaac cttccaagag
      481 caaaggagaa agtaaggaaa ttgacgccac caaagccttc tctcaactgg agattgctaa
      541 agcagagggc atggttagta aaggagaaga aaataacatg gcactgatta aggagaacat
      601 gcacatgaag ctgtacatgg agggcaccgt gaacaaccac cacttcaagt gcacatccga
      661 gggcgaaggc aagccctacg agggcaccca gaccatgaga atcaaggccg tcgagggcgg
      721 ccctctcccc ttcgccttcg acatcctggc taccagcttc atgtacggca gcaaaacctt
      781 catcaaccac acccagggca tccccgactt ctttaagcag tccttccctg agggcttcac
      841 atgggagaga gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag
      901 cctccaggac ggctgcctca tctacaacgt caagatcaga ggggtgaact tcccatccaa
      961 cggccctgtg atgcagaaga aaacactcgg ctgggaggcc tccaccgaga ccctgtaccc
     1021 cgctgacggc ggcctggaag gcagagccga catggccctg aagctcgtgg gcgggggcca
     1081 cctgatctgc aacttgaaga ccacatacag atccaagaaa cccgctaaga acctcaagat
     1141 gcccggcgtc tactatgtgg acagaagact ggaaagaatc aaggaggccg acaaagagac
     1201 ctacgtcgag cagcacgagg tggctgtggc cagatactgc gacctcccta gcaaactggg
     1261 gcacagagtc taataatcgc tgggacgccc ggcggccgcg gaaacacaga aaaaagcccg
     1321 cacctgacag tgcgggcttt ttttttcgac caaaggtagc gaacgacgag tcactgttga
     1381 ggataaatac tttctctact aggcgcctgt tacacaggtc ctcagcggcg cgcctgctgc
     1441 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     1501 tttgccaggc atcaaataaa acgaaaggct cagtcggaag actgggcctt tcgttttatc
     1561 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
     1621 gttgtgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat
     1681 caaactaagc agaaggccat cctgacggat ggcctttttg cgtttcagat ctaccggtaa
     1741 accagcaata gacataagcg gctatttaac gaccctgccc tgaaccgacg accgggtcat
     1801 cgtggccgga tcttgcggcc cctcggcttg aacgaattgt tagacattat ttgccgacta
     1861 ccttggtgat ctcgcctttc acgtagtgga caaattcttc caactgatct gcgcgcgagg
     1921 ccaagcgatc ttcttcttgt ccaagataag cctgtctagc ttcaagtatg acgggctgat
     1981 actgggccgg caggcgctcc attgcccagt cggcagcgac atccttcggc gcgattttgc
     2041 cggttactgc gctgtaccaa atgcgggaca acgtaagcac tacatttcgc tcatcgccag
     2101 cccagtcggg cggcgagttc catagcgtta aggtttcatt tagcgcctca aatagatcct
     2161 gttcaggaac cggatcaaag agttcctccg ccgctggacc taccaaggca acgctatgtt
     2221 ctcttgcttt tgtcagcaag atagccagat caatgtcgat cgtggctggc tcgaagatac
     2281 ctgcaagaat gtcattgcgc tgccattctc caaattgcag ttcgcgctta gctggataac
     2341 gccacggaat gatgtcgtcg tgcacaacaa tggtgacttc tacagcgcgg agaatctcgc
     2401 tctctccagg ggaagccgaa gtttccaaaa ggtcgttgat caaagctcgc cgcgttgttt
     2461 catcaagcct tacggtcacc gtaaccagca aatcaatatc actgtgtggc ttcaggccgt
     2521 catccactgc ggagccgtac aaatgtacgg ccagcaacgt cggttcgaga tggcgctcga
     2581 tgacgccaac tacctctgat agttgagtcg atacttcggc gatcaccgct tccctcatac
     2641 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     2701 tatttgaatg tatttagaaa aataaacaaa tagctagctc actcggtcgc tacgctccgg
     2761 gcgtgagact gcggcgggcg ctgcggacac atacaaagtt acccacagat tccgtggata
     2821 agcaggggac taacatgtga ggcaaaacag cagggccgcg ccggtggcgt ttttccatag
     2881 gctccgccct cctgccagag ttcacataaa cagacgcttt tccggtgcat ctgtgggagc
     2941 cgtgaggctc aaccatgaat ctgacagtac gggcgaaacc cgacaggact taaagatccc
     3001 caccgtttcc ggcgggtcgc tccctcttgc gctctcctgt tccgaccctg ccgtttaccg
     3061 gatacctgtt ccgcctttct cccttacggg aagtgtggcg ctttctcata gctcacacac
     3121 tggtatctcg gctcggtgta ggtcgtttgc tccaagctgg gctgtaagca agaactcccc
     3181 gttcagcccg actgctgcgc cttatccggt aactgttcac ttgagtccaa cccggaaaag
     3241 cacggtaaaa cgccactggc agcagccatt ggtaactggg agttcgcaga ggatttgttt
     3301 agctaaacac gcggttgctc ttgaagtgtg cgccaaagtc cggctacact ggaaggacag
     3361 atttggttgc tgtgctctgc gaaagccagt taccacggtt aagcagttcc ccaactgact
     3421 taaccttcga tcaaaccacc tccccaggtg gttttttcgt ttacagggca aaagattacg
     3481 cgcagaaaaa aaggatctca agaagatcct ttgatc
//
