LOCUS       sequence-124858-        4542 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pSTC2,TheoHHAzRaj12,cisRaj12-sfGFP, kanR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4542)
  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 4542)
  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: pSCKtheoRaj12
FEATURES             Location/Qualifiers
     source          1..4542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..22
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    227..245
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    252..270
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    315..331
                     /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             378..389
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             405..1115
                     /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      1202..1233
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(1332..2147)
                     /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     2055..2074
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2139..2158
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(2505..3455)
                     /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      3503..3725
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(4217..4311)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      complement(4467..4496)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 gaattcgcgg ccgcttctag agacagaaaa gcccgccttt cggcgggctt tgcccagaat
       61 cccttcgagc aaatcctagc gaggtgccgc tagattcgct caaaggaacc cttcttcctg
      121 cccgtctgga atccaggaca cctgccaagg gcctttcggc tggtatgtat ttcgtcctat
      181 ttgggactca tcagctggat gtacgaaggc agcgccgtgc tcagtatctc tatcactgat
      241 agggatgtca atctctatca ctgataggga gctagccaat tgatgcatat aaatgtgagc
      301 ggataacatt gacattgtga gcggataaca agatactgag cacacccagt atcattctct
      361 tcttcctgcc cacgcggaaa gaggagaaag gtgtaatgac tagtcgtaaa ggcgaagagc
      421 tgttcactgg tgtcgtccct attctggtgg aactggatgg tgatgtcaac ggtcataagt
      481 tttccgtgcg tggcgagggt gaaggtgacg caactaatgg taaactgacg ctgaagttca
      541 tctgtactac tggtaaactg ccggtacctt ggccgactct ggtaacgacg ctgacttatg
      601 gtgttcagtg ctttgctcgt tatccggacc atatgaagca gcatgacttc ttcaagtccg
      661 ccatgccgga aggctatgtg caggaacgca cgatttcctt taaggatgac ggcacgtaca
      721 aaacgcgtgc ggaagtgaaa tttgaaggcg ataccctggt aaaccgcatt gagctgaaag
      781 gcattgactt taaagaagac ggcaatatcc tgggccataa gctggaatac aattttaaca
      841 gccacaatgt ttacatcacc gccgataaac aaaaaaatgg cattaaagcg aattttaaaa
      901 ttcgccacaa cgtggaggat ggcagcgtgc agctggctga tcactaccag caaaacactc
      961 caatcggtga tggtcctgtt ctgctgccag acaatcacta tctgagcacg caaagcgttc
     1021 tgtctaaaga tccgaacgag aaacgcgatc atatggttct gctggagttc gtaaccgcag
     1081 cgggcatcac gcatggtatg gatgaactgt acaaagctag cgcagcgaac gacgaaaatt
     1141 acgcccttgc agcgtaataa ctgcaggagt cactaagggt tagttagtta gattagcaga
     1201 aagtcaaaag cctccgaccg gaggcttttg actaaaactt cccttggggt tatcattggg
     1261 gctcactcaa aggcggtaat cagataaaaa aaatccttag ctttcgctaa ggatgatttc
     1321 tgctagtatt attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
     1381 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
     1441 aggcagttcc aaagaatggc aaggtcctgg taacggtctg cgattccgac ccgtccaaca
     1501 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
     1561 tgagtgacga ctgaatccgg tgagaatggc aagagcttgt gcatttcttt ccagacttgt
     1621 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
     1681 atgcgtgatt gcgcctgagc aagacgaaat acacgatcgc tgttaaaagg acaattacaa
     1741 acaggaatcg aatgtaaccg gcgcaggaac acggccagcg catcaacaat attttcacct
     1801 gaatcaggat attcttctaa tacctggaag gctgttttcc caggaatcgc ggtggtgagt
     1861 aaccacgcat catcaggagt acggataaaa tgcttgatgg tcgggagagg cataaactcc
     1921 gtcagccagt tgagacggac catctcatct gtaacatcat tggcaacgct acctttgcca
     1981 tgtttcagaa acaactctgg cgcatcgggc ttcccataca agcgatagat tgtcgcacct
     2041 gattgcccga cattatcgcg agcccattta tacccatata aatcagcgtc catgttggag
     2101 tttaagcgcg gacgggagca agacgtttcc cgttgaatat ggctcataac accccttgta
     2161 ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca
     2221 atgtaacatc agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct
     2281 gagttgaagg atcagctcta gtagttacat tgtcgatctg ttcatggtga acagctttaa
     2341 atgcaccaaa aactcgtaaa agctctgatg tatctatctt ttttacaccg ttttcatctg
     2401 tgcatatgga cagttttccc tttgatatct aacggtgaac agttgttcta cttttgtttg
     2461 ttagtcttga tgcttcactg atagatacaa gagccataag aacctcagat ccttccgtat
     2521 ttagccagta tgttctctag tgtggttcgt tgtttttgcg tgagccatga gaacgaacca
     2581 ttgagatcat gcttactttg catgtcactc aaaaattttg cctcaaaact ggtgagctga
     2641 atttttgcag ttaaagcatc gtgtagtgtt tttcttagtc cgttacgtag gtaggaatct
     2701 gatgtaatgg ttgttggtat tttgtcacca ttcattttta tctggttgtt ctcaagttcg
     2761 gttacgagat ccatttgtct atctagttca acttggaaaa tcaacgtatc agtcgggcgg
     2821 cctcgcttat caaccaccaa tttcatattg ctgtaagtgt ttaaatcttt acttattggt
     2881 ttcaaaaccc attggttaag ccttttaaac tcatggtagt tattttcaag cattaacatg
     2941 aacttaaatt catcaaggct aatctctata tttgccttgt gagttttctt ttgtgttagt
     3001 tcttttaata accactcata aatcctcata gagtatttgt tttcaaaaga cttaacatgt
     3061 tccagattat attttatgaa tttttttaac tggaaaagat aaggcaatat ctcttcacta
     3121 aaaactaatt ctaatttttc gcttgagaac ttggcatagt ttgtccactg gaaaatctca
     3181 aagcctttaa ccaaaggatt cctgatttcc acagttctcg tcatcagctc tctggttgct
     3241 ttagctaata caccataagc attttcccta ctgatgttca tcatctgagc gtattggtta
     3301 taagtgaacg ataccgtccg ttctttcctt gtagggtttt caatcgtggg gttgagtagt
     3361 gccacacagc ataaaattag cttggtttca tgctccgtta agtcatagcg actaatcgct
     3421 agttcatttg ctttgaaaac aactaattca gacatacatc tcaattggtc taggtgattt
     3481 taatcactat accaattgag atgggctagt caatgataat tacatgtcct tttcctttga
     3541 gttgtgggta tctgtaaatt ctgctagacc tttgctggaa aacttgtaaa ttctgctaga
     3601 ccctctgtaa attccgctag acctttgtgt gttttttttg tttatattca agtggttata
     3661 atttatagaa taaagaaaga ataaaaaaag ataaaaagaa tagatcccag ccctgtgtat
     3721 aactcactac tttagtcagt tccgcagtat tacaaaagga tgtcgcaaac gctgtttgct
     3781 cctctacaaa acagacctta aaaccctaaa ggcttaagta gcaccctcgc aagctcgggc
     3841 aaatcgctga atattccttt tgtctccgac catcaggcac ctgagtcgct gtctttttcg
     3901 tgacattcag ttcgctgcgc tcacggctct ggcagtgaat gggggtaaat ggcactacag
     3961 gcgcctttta tggattcatg caaggaaact acccataata caagaaaagc ccgtcacggg
     4021 cttctcaggg cgttttatgg cgggtctgct atgtggtgct atctgacttt ttgctgttca
     4081 gcagttcctg ccctctgatt ttccagtctg accacttcgg attatcccgt gacaggtcat
     4141 tcagactggc taatgcaccc agtaaggcag cggtatcatc aacaggctta cccgtcttac
     4201 tgtccctagt gcttggattc tcaccaataa aaaacgcccg gcggcaaccg agcgttctga
     4261 acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt ccaagcgagc
     4321 tcgtaaactt ggtctgacag ctctagctcc ggcaaaaaaa cgggcaaggt gtcaccaccc
     4381 tgcccttttt ctttaaaacc gaaaagatta cttcgcgttt gccacctgac gtctaagaaa
     4441 aggaatattc agcaatttgc ccgtgccgaa gaaaggccca cccgtgaagg tgagccagtg
     4501 agttgattgc tacgtaatta gttagttagc ccttagtgac tc
//
