LOCUS       sequence-327547-        4024 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains elements for building reporter plasmid for IBM.TetR 
            reporter plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4024)
  AUTHORS   Ho TYH, Shao A, Lu Z, Savilahti H, Menolascina F, Wang L, Dalchau N,
            Wang B
  TITLE     A systematic approach to inserting split inteins for Boolean logic 
            gate engineering and basal activity reduction.
  JOURNAL   Nat Commun. 2021 Apr 13;12(1):2200. doi: 10.1038/s41467-021-22404-9.
  PUBMED    33850130
REFERENCE   2  (bases 1 to 4024)
  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"; journalName: "Nat 
            Commun."; date: "2021-04-13"; volume: "12(1):2200. doi"; pages: " 
            10.1038/s41467-021-22404-9"
COMMENT     Sequence Label: IBMc259
FEATURES             Location/Qualifiers
     source          1..4024
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             186..977
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     oriT            1123..1231
                     /note="incP origin of transfer"
     CDS             complement(1242..2192)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFDIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2240..2462
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(2729..2815)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2830..2852
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    2877..2898
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    2940..2958
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2965..2983
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    3855..3875
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      3880..3974
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttg
       61 gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt
      121 tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttaa
      181 acatcatgag ggaagcggtg atcgccgaag tatcgactca actatcagag gtagttggcg
      241 tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg
      301 atggcggcct gaagccacac agtgatattg atttgctggt tacggtgacc gtaaggcttg
      361 atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct tcccctggag
      421 agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac atcattccgt
      481 ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg
      541 caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa
      601 gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg
      661 aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact
      721 gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa
      781 ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc
      841 agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct
      901 tggcctcgcg cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca
      961 aggtagtcgg caaataagac aattgtcctt ttccgctgca taaccctgct tcggggtcat
     1021 tatagcgatt ttttcggtat atccatcctt tttcgcacga tatacaggat tttgccaaag
     1081 ggttcgtgta gactttcctt ggtgtatcca acggcgtcag ccgggcagga taggtgaagt
     1141 aggcccaccc gcgagcgggt gttccttctt cactgtccct tattcgcacc tggcggtgct
     1201 caacgggaat cctgctctgc gaggctggcc gtaggccggc ctcagatcct tccgtattta
     1261 gccagtatgt tctctagtgt ggttcgttgt ttttgcgtga gccatgagaa cgaaccattg
     1321 agatcatact tactttgcat gtcactcaaa aattttgcct caaaactggt gagctgaatt
     1381 tttgcagtta aagcatcgtg tagtgttttt cttagtccgt tatgtaggta ggaatctgat
     1441 gtaatggttg ttggtatttt gtcaccattc atttttatct ggttgttctc aagttcggtt
     1501 acgagatcca tttgtctatc tagttcaact tggaaaatca acgtatcagt cgggcggcct
     1561 cgcttatcaa ccaccaattt catattgctg taagtgttta aatctttact tattggtttc
     1621 aaaacccatt ggttaagcct tttaaactca tggtagttat tttcaagcat taacatgaac
     1681 ttaaattcat caaggctaat ctctatattt gccttgtgag ttttcttttg tgttagttct
     1741 tttaataacc actcataaat cctcatagag tatttgtttt caaaagactt aacatgttcc
     1801 agattatatt ttatgaattt ttttaactgg aaaagataag gcaatatctc ttcactaaaa
     1861 actaattcta atttttcgct tgagaacttg gcatagtttg tccactggaa aatatcaaag
     1921 cctttaacca aaggattcct gatttccaca gttctcgtca tcagctctct ggttgcttta
     1981 gctaatacac cataagcatt ttccctactg atgttcatca tctgagcgta ttggttataa
     2041 gtgaacgata ccgtccgttc tttccttgta gggttttcaa tcgtggggtt gagtagtgcc
     2101 acacagcata aaattagctt ggtttcatgc tccgttaagt catagcgact aatcgctagt
     2161 tcatttgctt tgaaaacaac taattcagac atacatctca attggtctag gtgattttaa
     2221 tcactatacc aattgagatg ggctagtcaa tgataattac atgtcctttt cctttgagtt
     2281 gtgggtatct gtaaattctg ctagaccttt gctggaaaac ttgtaaattc tgctagaccc
     2341 tctgtaaatt ccgctagacc tttgtgtgtt ttttttgttt atattcaagt ggttataatt
     2401 tatagaataa agaaagaata aaaaaagata aaaagaatag atcccagccc tgtgtataac
     2461 tcactacttt agtcagttcc gcagtattac aaaaggatgt cgcaaacgct gtttgctcct
     2521 ctacaaaaca gaccttaaaa ccctaaaggc ttaagtagca ccctcgcaag ctcgggcaaa
     2581 tcgctgaata ttccttttgt ctccgaccat caggcacctg agtcgctgtc tttttcgtga
     2641 cattcagttc gctgcgctca cggctctggc agtgaatggg ggtaaatggc actacaggcg
     2701 cggcgcgccc agctgtctag ggcggcggat ttgtcctact caggagagcg ttcaccgaca
     2761 aacaacagat aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt atttgatgcc
     2821 tttaattaaa gcggataaca atttcacaca ggaggccgcc taggccgcgg ccgcgcgaat
     2881 tcgcggccgc ttctagaggc ttaacgatcg ttggctgttt tcagcaggac gcactgacct
     2941 ccctatcagt gatagagatt gacatcccta tcagtgatag agatactgag cacagcgctc
     3001 aacgggtgtg cttcccgttc tgatgagtcc gtgaggacga aagcgcctct acaaataatt
     3061 ttgtttaata ctagagaaag aggggaaata ctagatggtg agtaaaggag aagctgtgat
     3121 taaagagttc atgcgcttca aagttcacat ggagggttct atgaacggtc acgagttcga
     3181 gatcgaaggc gaaggcgagg gccgtccgta tgaaggcacc cagaccgcca aactgaaagt
     3241 gactaaaggc ggcccgctgc ctttttcctg ggacatcctg agcccgcaat ttatgtacgg
     3301 ttctagggcg ttcatcaaac acccagcgga tatcccggac tattataagc agtcttttcc
     3361 ggaaggtttc aagtgggaac gcgtaatgaa ttttgaagat ggtggtgccg tgaccgtcac
     3421 tcaggacacc tccctggagg atggcaccct gatctataaa gttaaactgc gtggtactaa
     3481 ttttccacct gatggcccgg tgatgcagaa aaagacgatg ggttgggagg cgtctaccga
     3541 acgcttgtat ccggaagatg gtgtgctgaa aggcgacatt aaaatggccc tgcgcctgaa
     3601 agatggcggc cgctatctgg ctgacttcaa aaccacgtac aaagccaaga aacctgtgca
     3661 gatgcctggc gcgtacaatg tggaccgcaa actggacatc acctctcata atgaagatta
     3721 tacggtggta gagcaatatg agcgctccga gggtcgtcat tctaccggtg gcatggatga
     3781 actatacaaa taactcggta ccaaattcca gaaaagaggc ctcccgaaag gggggccttt
     3841 tttcgttttg gtcctactag tagcggccgc tgcagcttgg actcctgttg atagatccag
     3901 taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt
     3961 ttattggtga gaatccaggg gtccccaata attacgattt acgtatttaa atgaaccttg
     4021 accg
//
