LOCUS       sequence-124981-        4524 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pSTC2,theoHHAzRaj12,cisRaj11-sfGFP, KanR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4524)
  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 4524)
  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-o11
FEATURES             Location/Qualifiers
     source          1..4524
                     /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    321..337
                     /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             384..395
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             411..1121
                     /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"
     misc_feature    1128..1148
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1184..1215
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(1314..2129)
                     /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     2037..2056
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2121..2140
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(2487..3437)
                     /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      3485..3707
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(4199..4293)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      complement(4449..4478)
                     /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 tgatgcatgc tagcataaat
      301 gtgagcggat aacattgaca ttgtgagcgg ataacaagat actgagcacc ctcgcataat
      361 ttcacttctt caatcctccc gttaaagagg agaaattatg aatgactagc cgtaaaggcg
      421 aagagctgtt cactggtgtc gtccctattc tggtggaact ggatggtgat gtcaacggtc
      481 ataagttttc cgtgcgtggc gagggtgaag gtgacgcaac taatggtaaa ctgacgctga
      541 agttcatctg tactactggt aaactgccgg taccttggcc gactctggta acgacgctga
      601 cttatggtgt tcagtgcttt gctcgttatc cggaccatat gaagcagcat gacttcttca
      661 agtccgccat gccggaaggc tatgtgcagg aacgcacgat ttcctttaag gatgacggca
      721 cgtacaaaac gcgtgcggaa gtgaaatttg aaggcgatac cctggtaaac cgcattgagc
      781 tgaaaggcat tgactttaaa gaagacggca atatcctggg ccataagctg gaatacaatt
      841 ttaacagcca caatgtttac atcaccgccg ataaacaaaa aaatggcatt aaagcgaatt
      901 ttaaaattcg ccacaacgtg gaggatggca gcgtgcagct ggctgatcac taccagcaaa
      961 acactccaat cggtgatggt cctgttctgc tgccagacaa tcactatctg agcacgcaaa
     1021 gcgttctgtc taaagatccg aacgagaaac gcgatcatat ggttctgctg gagttcgtaa
     1081 ccgcagcggg catcacgcat ggtatggatg aactgtacaa atgatgatac tagtagcggc
     1141 cgctgcagga gtcactaagg gttagttagt tagattagca gaaagtcaaa agcctccgac
     1201 cggaggcttt tgactaaaac ttcccttggg gttatcattg gggctcactc aaaggcggta
     1261 atcagataaa aaaaatcctt agctttcgct aaggatgatt tctgctagta ttattagaaa
     1321 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
     1381 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccaaagaatg
     1441 gcaaggtcct ggtaacggtc tgcgattccg acccgtccaa catcaataca acctattaat
     1501 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     1561 ggtgagaatg gcaagagctt gtgcatttct ttccagactt gttcaacagg ccagccatta
     1621 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcatgcgtga ttgcgcctga
     1681 gcaagacgaa atacacgatc gctgttaaaa ggacaattac aaacaggaat cgaatgtaac
     1741 cggcgcagga acacggccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     1801 aatacctgga aggctgtttt cccaggaatc gcggtggtga gtaaccacgc atcatcagga
     1861 gtacggataa aatgcttgat ggtcgggaga ggcataaact ccgtcagcca gttgagacgg
     1921 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     1981 ggcgcatcgg gcttcccata caagcgatag attgtcgcac ctgattgccc gacattatcg
     2041 cgagcccatt tatacccata taaatcagcg tccatgttgg agtttaagcg cggacgggag
     2101 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
     2161 gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt
     2221 tgagacacaa cgtggctttg ttgaataaat cgaacttttg ctgagttgaa ggatcagctc
     2281 tagtagttac attgtcgatc tgttcatggt gaacagcttt aaatgcacca aaaactcgta
     2341 aaagctctga tgtatctatc ttttttacac cgttttcatc tgtgcatatg gacagttttc
     2401 cctttgatat ctaacggtga acagttgttc tacttttgtt tgttagtctt gatgcttcac
     2461 tgatagatac aagagccata agaacctcag atccttccgt atttagccag tatgttctct
     2521 agtgtggttc gttgtttttg cgtgagccat gagaacgaac cattgagatc atgcttactt
     2581 tgcatgtcac tcaaaaattt tgcctcaaaa ctggtgagct gaatttttgc agttaaagca
     2641 tcgtgtagtg tttttcttag tccgttacgt aggtaggaat ctgatgtaat ggttgttggt
     2701 attttgtcac cattcatttt tatctggttg ttctcaagtt cggttacgag atccatttgt
     2761 ctatctagtt caacttggaa aatcaacgta tcagtcgggc ggcctcgctt atcaaccacc
     2821 aatttcatat tgctgtaagt gtttaaatct ttacttattg gtttcaaaac ccattggtta
     2881 agccttttaa actcatggta gttattttca agcattaaca tgaacttaaa ttcatcaagg
     2941 ctaatctcta tatttgcctt gtgagttttc ttttgtgtta gttcttttaa taaccactca
     3001 taaatcctca tagagtattt gttttcaaaa gacttaacat gttccagatt atattttatg
     3061 aattttttta actggaaaag ataaggcaat atctcttcac taaaaactaa ttctaatttt
     3121 tcgcttgaga acttggcata gtttgtccac tggaaaatct caaagccttt aaccaaagga
     3181 ttcctgattt ccacagttct cgtcatcagc tctctggttg ctttagctaa tacaccataa
     3241 gcattttccc tactgatgtt catcatctga gcgtattggt tataagtgaa cgataccgtc
     3301 cgttctttcc ttgtagggtt ttcaatcgtg gggttgagta gtgccacaca gcataaaatt
     3361 agcttggttt catgctccgt taagtcatag cgactaatcg ctagttcatt tgctttgaaa
     3421 acaactaatt cagacataca tctcaattgg tctaggtgat tttaatcact ataccaattg
     3481 agatgggcta gtcaatgata attacatgtc cttttccttt gagttgtggg tatctgtaaa
     3541 ttctgctaga cctttgctgg aaaacttgta aattctgcta gaccctctgt aaattccgct
     3601 agacctttgt gtgttttttt tgtttatatt caagtggtta taatttatag aataaagaaa
     3661 gaataaaaaa agataaaaag aatagatccc agccctgtgt ataactcact actttagtca
     3721 gttccgcagt attacaaaag gatgtcgcaa acgctgtttg ctcctctaca aaacagacct
     3781 taaaacccta aaggcttaag tagcaccctc gcaagctcgg gcaaatcgct gaatattcct
     3841 tttgtctccg accatcaggc acctgagtcg ctgtcttttt cgtgacattc agttcgctgc
     3901 gctcacggct ctggcagtga atgggggtaa atggcactac aggcgccttt tatggattca
     3961 tgcaaggaaa ctacccataa tacaagaaaa gcccgtcacg ggcttctcag ggcgttttat
     4021 ggcgggtctg ctatgtggtg ctatctgact ttttgctgtt cagcagttcc tgccctctga
     4081 ttttccagtc tgaccacttc ggattatccc gtgacaggtc attcagactg gctaatgcac
     4141 ccagtaaggc agcggtatca tcaacaggct tacccgtctt actgtcccta gtgcttggat
     4201 tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt
     4261 ctgaggtcat tactggatct atcaacagga gtccaagcga gctcgtaaac ttggtctgac
     4321 agctctagct ccggcaaaaa aacgggcaag gtgtcaccac cctgcccttt ttctttaaaa
     4381 ccgaaaagat tacttcgcgt ttgccacctg acgtctaaga aaaggaatat tcagcaattt
     4441 gcccgtgccg aagaaaggcc cacccgtgaa ggtgagccag tgagttgatt gctacgtaat
     4501 tagttagtta gcccttagtg actc
//
