LOCUS       sequence-124987-        4655 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pSTC2,cisRaj12-sfGFP, KanR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4655)
  AUTHORS   Shen S, Rodrigo G, Prakash S, Majer E, Landrain TE, Kirov B, Daros 
            JA, Jaramillo A
  TITLE     Dynamic signal processing by ribozyme-mediated RNA circuits to 
            control gene expression.
  JOURNAL   Nucleic Acids Res. 2015 May 26;43(10):5158-70. doi: 
            10.1093/nar/gkv287. Epub 2015 Apr 27.
  PUBMED    25916845
REFERENCE   2  (bases 1 to 4655)
  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: 
            "10.1093/nar/gkv287"; journalName: "Nucleic Acids Res"; date: 
            "2015-05-26- 26"; volume: "43"; issue: "10"; pages: "5158-70"
COMMENT     Sequence Label: pSCKbreak12
FEATURES             Location/Qualifiers
     source          1..4655
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    83..101
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    108..126
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        139..173
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     protein_bind    428..444
                     /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)."
     RBS             491..502
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             518..1228
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="RKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     terminator      1315..1346
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(1445..2260)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2168..2187
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2252..2271
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(2618..3568)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      3616..3838
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(4330..4424)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      complement(4580..4609)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 gaattctcta gaaaaaaaaa tccttagctt tcgctaagga tagctcgtca ctgtccaggt
       61 tcagggtttc ccgtgctcag tatctctatc actgataggg atgtcaatct ctatcactga
      121 tagggatcta gaatcgattt gacagctagc tcagtcctag gtataatgct agcggcgctg
      181 ccttcgggcg accctgatga gcttgagttt agctcgtcac tgtccaggtt caatcaggcg
      241 aaacggtgaa agccgtaggt tgcccgacgg gcaggaagaa gggttccttt gagcgaatct
      301 agcggcacct cgctaggatt tgctcgaagg gattctgggc aaagcccgcc gaaaggcggg
      361 cttttctgta ctagtctgca gggatcgacg agagcatgca tataaatgtg agcggataac
      421 attgacattg tgagcggata acaagatact gagcacaccc agtatcattc tcttcttcct
      481 gcccacgcgg aaagaggaga aaggtgtaat gactagtcgt aaaggcgaag agctgttcac
      541 tggtgtcgtc cctattctgg tggaactgga tggtgatgtc aacggtcata agttttccgt
      601 gcgtggcgag ggtgaaggtg acgcaactaa tggtaaactg acgctgaagt tcatctgtac
      661 tactggtaaa ctgccggtac cttggccgac tctggtaacg acgctgactt atggtgttca
      721 gtgctttgct cgttatccgg accatatgaa gcagcatgac ttcttcaagt ccgccatgcc
      781 ggaaggctat gtgcaggaac gcacgatttc ctttaaggat gacggcacgt acaaaacgcg
      841 tgcggaagtg aaatttgaag gcgataccct ggtaaaccgc attgagctga aaggcattga
      901 ctttaaagaa gacggcaata tcctgggcca taagctggaa tacaatttta acagccacaa
      961 tgtttacatc accgccgata aacaaaaaaa tggcattaaa gcgaatttta aaattcgcca
     1021 caacgtggag gatggcagcg tgcagctggc tgatcactac cagcaaaaca ctccaatcgg
     1081 tgatggtcct gttctgctgc cagacaatca ctatctgagc acgcaaagcg ttctgtctaa
     1141 agatccgaac gagaaacgcg atcatatggt tctgctggag ttcgtaaccg cagcgggcat
     1201 cacgcatggt atggatgaac tgtacaaagc tagcgcagcg aacgacgaaa attacgccct
     1261 tgcagcgtaa taactgcagg agtcactaag ggttagttag ttagattagc agaaagtcaa
     1321 aagcctccga ccggaggctt ttgactaaaa cttcccttgg ggttatcatt ggggctcact
     1381 caaaggcggt aatcagataa aaaaaatcct tagctttcgc taaggatgat ttctgctagt
     1441 attattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     1501 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     1561 tccaaagaat ggcaaggtcc tggtaacggt ctgcgattcc gacccgtcca acatcaatac
     1621 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     1681 cgactgaatc cggtgagaat ggcaagagct tgtgcatttc tttccagact tgttcaacag
     1741 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcatgcgtg
     1801 attgcgcctg agcaagacga aatacacgat cgctgttaaa aggacaatta caaacaggaa
     1861 tcgaatgtaa ccggcgcagg aacacggcca gcgcatcaac aatattttca cctgaatcag
     1921 gatattcttc taatacctgg aaggctgttt tcccaggaat cgcggtggtg agtaaccacg
     1981 catcatcagg agtacggata aaatgcttga tggtcgggag aggcataaac tccgtcagcc
     2041 agttgagacg gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     2101 gaaacaactc tggcgcatcg ggcttcccat acaagcgata gattgtcgca cctgattgcc
     2161 cgacattatc gcgagcccat ttatacccat ataaatcagc gtccatgttg gagtttaagc
     2221 gcggacggga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     2281 ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac
     2341 atcagagatt ttgagacaca acgtggcttt gttgaataaa tcgaactttt gctgagttga
     2401 aggatcagct ctagtagtta cattgtcgat ctgttcatgg tgaacagctt taaatgcacc
     2461 aaaaactcgt aaaagctctg atgtatctat cttttttaca ccgttttcat ctgtgcatat
     2521 ggacagtttt ccctttgata tctaacggtg aacagttgtt ctacttttgt ttgttagtct
     2581 tgatgcttca ctgatagata caagagccat aagaacctca gatccttccg tatttagcca
     2641 gtatgttctc tagtgtggtt cgttgttttt gcgtgagcca tgagaacgaa ccattgagat
     2701 catgcttact ttgcatgtca ctcaaaaatt ttgcctcaaa actggtgagc tgaatttttg
     2761 cagttaaagc atcgtgtagt gtttttctta gtccgttacg taggtaggaa tctgatgtaa
     2821 tggttgttgg tattttgtca ccattcattt ttatctggtt gttctcaagt tcggttacga
     2881 gatccatttg tctatctagt tcaacttgga aaatcaacgt atcagtcggg cggcctcgct
     2941 tatcaaccac caatttcata ttgctgtaag tgtttaaatc tttacttatt ggtttcaaaa
     3001 cccattggtt aagcctttta aactcatggt agttattttc aagcattaac atgaacttaa
     3061 attcatcaag gctaatctct atatttgcct tgtgagtttt cttttgtgtt agttctttta
     3121 ataaccactc ataaatcctc atagagtatt tgttttcaaa agacttaaca tgttccagat
     3181 tatattttat gaattttttt aactggaaaa gataaggcaa tatctcttca ctaaaaacta
     3241 attctaattt ttcgcttgag aacttggcat agtttgtcca ctggaaaatc tcaaagcctt
     3301 taaccaaagg attcctgatt tccacagttc tcgtcatcag ctctctggtt gctttagcta
     3361 atacaccata agcattttcc ctactgatgt tcatcatctg agcgtattgg ttataagtga
     3421 acgataccgt ccgttctttc cttgtagggt tttcaatcgt ggggttgagt agtgccacac
     3481 agcataaaat tagcttggtt tcatgctccg ttaagtcata gcgactaatc gctagttcat
     3541 ttgctttgaa aacaactaat tcagacatac atctcaattg gtctaggtga ttttaatcac
     3601 tataccaatt gagatgggct agtcaatgat aattacatgt ccttttcctt tgagttgtgg
     3661 gtatctgtaa attctgctag acctttgctg gaaaacttgt aaattctgct agaccctctg
     3721 taaattccgc tagacctttg tgtgtttttt ttgtttatat tcaagtggtt ataatttata
     3781 gaataaagaa agaataaaaa aagataaaaa gaatagatcc cagccctgtg tataactcac
     3841 tactttagtc agttccgcag tattacaaaa ggatgtcgca aacgctgttt gctcctctac
     3901 aaaacagacc ttaaaaccct aaaggcttaa gtagcaccct cgcaagctcg ggcaaatcgc
     3961 tgaatattcc ttttgtctcc gaccatcagg cacctgagtc gctgtctttt tcgtgacatt
     4021 cagttcgctg cgctcacggc tctggcagtg aatgggggta aatggcacta caggcgcctt
     4081 ttatggattc atgcaaggaa actacccata atacaagaaa agcccgtcac gggcttctca
     4141 gggcgtttta tggcgggtct gctatgtggt gctatctgac tttttgctgt tcagcagttc
     4201 ctgccctctg attttccagt ctgaccactt cggattatcc cgtgacaggt cattcagact
     4261 ggctaatgca cccagtaagg cagcggtatc atcaacaggc ttacccgtct tactgtccct
     4321 agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc
     4381 cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagcg agctcgtaaa
     4441 cttggtctga cagctctagc tccggcaaaa aaacgggcaa ggtgtcacca ccctgccctt
     4501 tttctttaaa accgaaaaga ttacttcgcg tttgccacct gacgtctaag aaaaggaata
     4561 ttcagcaatt tgcccgtgcc gaagaaaggc ccacccgtga aggtgagcca gtgagttgat
     4621 tgctacgtaa ttagttagtt agcccttagt gactc
//
