LOCUS       sequence-244527-        3494 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Slot B, contains sfGFP-MarAn20 (step 1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3494)
  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 3494)
  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-B-010
FEATURES             Location/Qualifiers
     source          1..3494
                     /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..1056
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1421..1439)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1435..1482
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1491..1577
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1669..1696
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(1825..2616)
                     /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(2617..2708)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2756..3494)
                     /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 cctgaattcg aaatccctga aactgagact gtagaaaata agcttgtcca gactattgga
      241 tccaagagat ttctacacga ttgagcactg tctcagcctg cggtccgggt tcactgccgt
      301 ataggcagta attttgttta actttaagaa ggagatatac atatgcgtaa aggcgaagaa
      361 ctgtttaccg gtgtggttcc gattctggtg gaactggacg gcgatgttaa tggtcataaa
      421 ttcagtgttc gcggcgaagg tgaaggcgat gcgacgaacg gcaaactgac cctgaaattt
      481 atctgcacca cgggtaaact gccggtcccg tggccgacgc tggtgaccac gctgacctat
      541 ggcgttcaat gttttgcgcg ttacccggat cacatgaaac agcacgactt tttcaaatcg
      601 gccatgccgg aaggctatgt gcaggaacgt acgattagct ttaaagacga tggtacgtat
      661 aaaacccgcg cggaagtgaa attcgaaggc gataccctgg ttaaccgtat cgaactgaaa
      721 ggtatcgatt tcaaagaaga cggcaatatt ctgggtcata aactggaata taacttcaat
      781 tcccacaacg tgtacatcac cgcggataaa cagaaaaacg gcattaaagc caatttcaaa
      841 atccgccata atgtggaaga tggtagcgtt cagctggccg accactatca gcaaaacacg
      901 ccgattggtg atggcccggt cctgctgccg gacaatcact acctgagtac ccagtccgtg
      961 ctgtcaaaag atccgaacga aaaacgtgac cacatggtcc tgctggaatt tgtgacggct
     1021 gcgggtatca cccacggcat ggacgaactg tataaaatgt cccgccgtaa tactgacgcc
     1081 atcacaatcc acagcatcct ggattggatt gaagactaat aatcgctggg acgcccgcct
     1141 gcaggctcgg taccaaattc cagaaaagag gcctcccgaa aggggggcct tttttcgttt
     1201 tggtcctaat agataaagga taggtctggt agtgttgttc gttctcgcag gtaaatcaat
     1261 aatactcagc agttccgtag acttttcagt gggacagggt agcgataaca gatagattgt
     1321 aataagacac agtaggtgct cgtagttgcg tgaagagaac cgctcaggaa atccagtcag
     1381 aagtattggt aatcgttgaa aactcagtcg actgctgcca ccgctgagca ataactagca
     1441 taaccccttg gggcctctaa acgggtcttg aggggttttt tgccaggcat caaataaaac
     1501 gaaaggctca gtcggaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     1561 tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgtgaagcaa cggcccggag
     1621 ggtggcgggc aggacgcccg ccataaactg ccaggcatca aactaagcag aaggccatcc
     1681 tgacggatgg cctttttgcg tttcagatct accggtaaac cagcaataga cataagcggc
     1741 tatttaacga ccctgccctg aaccgacgac cgggtcatcg tggccggatc ttgcggcccc
     1801 tcggcttgaa cgaattgtta gacattattt gccgactacc ttggtgatct cgcctttcac
     1861 gtagtggaca aattcttcca actgatctgc gcgcgaggcc aagcgatctt cttcttgtcc
     1921 aagataagcc tgtctagctt caagtatgac gggctgatac tgggccggca ggcgctccat
     1981 tgcccagtcg gcagcgacat ccttcggcgc gattttgccg gttactgcgc tgtaccaaat
     2041 gcgggacaac gtaagcacta catttcgctc atcgccagcc cagtcgggcg gcgagttcca
     2101 tagcgttaag gtttcattta gcgcctcaaa tagatcctgt tcaggaaccg gatcaaagag
     2161 ttcctccgcc gctggaccta ccaaggcaac gctatgttct cttgcttttg tcagcaagat
     2221 agccagatca atgtcgatcg tggctggctc gaagatacct gcaagaatgt cattgcgctg
     2281 ccattctcca aattgcagtt cgcgcttagc tggataacgc cacggaatga tgtcgtcgtg
     2341 cacaacaatg gtgacttcta cagcgcggag aatctcgctc tctccagggg aagccgaagt
     2401 ttccaaaagg tcgttgatca aagctcgccg cgttgtttca tcaagcctta cggtcaccgt
     2461 aaccagcaaa tcaatatcac tgtgtggctt caggccgtca tccactgcgg agccgtacaa
     2521 atgtacggcc agcaacgtcg gttcgagatg gcgctcgatg acgccaacta cctctgatag
     2581 ttgagtcgat acttcggcga tcaccgcttc cctcatactc ttcctttttc aatattattg
     2641 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     2701 taaacaaata gctagctcac tcggtcgcta cgctccgggc gtgagactgc ggcgggcgct
     2761 gcggacacat acaaagttac ccacagattc cgtggataag caggggacta acatgtgagg
     2821 caaaacagca gggccgcgcc ggtggcgttt ttccataggc tccgccctcc tgccagagtt
     2881 cacataaaca gacgcttttc cggtgcatct gtgggagccg tgaggctcaa ccatgaatct
     2941 gacagtacgg gcgaaacccg acaggactta aagatcccca ccgtttccgg cgggtcgctc
     3001 cctcttgcgc tctcctgttc cgaccctgcc gtttaccgga tacctgttcc gcctttctcc
     3061 cttacgggaa gtgtggcgct ttctcatagc tcacacactg gtatctcggc tcggtgtagg
     3121 tcgtttgctc caagctgggc tgtaagcaag aactccccgt tcagcccgac tgctgcgcct
     3181 tatccggtaa ctgttcactt gagtccaacc cggaaaagca cggtaaaacg ccactggcag
     3241 cagccattgg taactgggag ttcgcagagg atttgtttag ctaaacacgc ggttgctctt
     3301 gaagtgtgcg ccaaagtccg gctacactgg aaggacagat ttggttgctg tgctctgcga
     3361 aagccagtta ccacggttaa gcagttcccc aactgactta accttcgatc aaaccacctc
     3421 cccaggtggt tttttcgttt acagggcaaa agattacgcg cagaaaaaaa ggatctcaag
     3481 aagatccttt gatc
//
