LOCUS       sequence-49410-e        4860 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4860)
  AUTHORS   Rodrigo G, Landrain TE, Jaramillo A
  TITLE     De novo automated design of small RNA circuits for engineering 
            synthetic riboregulation in living cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2012 Sep 18;109(38):15271-6. Epub 2012 Sep
            4.
  PUBMED    22949707
REFERENCE   2  (bases 1 to 4860)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2012-09-18"; volume: "109(38)"; pages: 
            "15271-6. Epub 2012 Sep 4"
COMMENT     Sequence Label: pSTC1
FEATURES             Location/Qualifiers
     source          1..4860
                     /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"""
     CDS             complement(26..703)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             710..721
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             747..1457
                     /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    1464..1484
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1520..1551
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(1650..2465)
                     /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     2373..2392
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2457..2476
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(2823..3773)
                     /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      3821..4043
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(4535..4629)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      complement(4785..4814)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 gaattcgcgg ccgcttctag agttattaag caccggtgga gtgacgacct tcagcacgtt
       61 cgtactgttc aacgatggtg tagtcttcgt tgtgggaggt gatgtccagt ttgatgtcgg
      121 ttttgtaagc acccggcagc tgaaccggtt ttttagccat gtaggtggtt ttaacttcag
      181 cgtcgtagtg accaccgtct ttcagtttca gacgcatttt gatttcacct ttcagagcac
      241 cgtcttccgg gtacatacgt tcggtggaag cttcccaacc catggttttt ttctgcataa
      301 ccggaccgtc ggacgggaag ttggtaccac gcagtttaac tttgtagatg aactcaccgt
      361 cttgcaggga ggagtcctgg gtaacggtaa caacaccacc gtcttcgaag ttcataacac
      421 gttcccattt gaaaccttcc gggaaggaca gtttcaggta gtccgggatg tcagccgggt
      481 gtttaacgta agctttggaa ccgtactgga actgcgggga caggatgtcc caagcgaacg
      541 gcagcggacc acctttggta actttcagtt tagcggtctg ggtaccttcg tacggacgac
      601 cttcaccttc accttcgatt tcgaactcgt gaccgttaac ggaaccttcc atacgaactt
      661 tgaaacgcat gaactctttg ataacgtctt cggaggaagc catctagtat ttctcctctt
      721 tctctagtgg atcccctagg gctagccgta aaggcgaaga gctgttcact ggtgtcgtcc
      781 ctattctggt ggaactggat ggtgatgtca acggtcataa gttttccgtg cgtggcgagg
      841 gtgaaggtga cgcaactaat ggtaaactga cgctgaagtt catctgtact actggtaaac
      901 tgccggtacc ttggccgact ctggtaacga cgctgactta tggtgttcag tgctttgctc
      961 gttatccgga ccatatgaag cagcatgact tcttcaagtc cgccatgccg gaaggctatg
     1021 tgcaggaacg cacgatttcc tttaaggatg acggcacgta caaaacgcgt gcggaagtga
     1081 aatttgaagg cgataccctg gtaaaccgca ttgagctgaa aggcattgac tttaaagaag
     1141 acggcaatat cctgggccat aagctggaat acaattttaa cagccacaat gtttacatca
     1201 ccgccgataa acaaaaaaat ggcattaaag cgaattttaa aattcgccac aacgtggagg
     1261 atggcagcgt gcagctggct gatcactacc agcaaaacac tccaatcggt gatggtcctg
     1321 ttctgctgcc agacaatcac tatctgagca cgcaaagcgt tctgtctaaa gatccgaacg
     1381 agaaacgcga tcatatggtt ctgctggagt tcgtaaccgc agcgggcatc acgcatggta
     1441 tggatgaact gtacaaatga tgatactagt agcggccgct gcaggagtca ctaagggtta
     1501 gttagttaga ttagcagaaa gtcaaaagcc tccgaccgga ggcttttgac taaaacttcc
     1561 cttggggtta tcattggggc tcactcaaag gcggtaatca gataaaaaaa atccttagct
     1621 ttcgctaagg atgatttctg ctagtattat tagaaaaact catcgagcat caaatgaaac
     1681 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     1741 gaaggagaaa actcaccgag gcagttccaa agaatggcaa ggtcctggta acggtctgcg
     1801 attccgaccc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     1861 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa gagcttgtgc
     1921 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     1981 tcaaccaaac cgttattcat gcgtgattgc gcctgagcaa gacgaaatac acgatcgctg
     2041 ttaaaaggac aattacaaac aggaatcgaa tgtaaccggc gcaggaacac ggccagcgca
     2101 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaaggc tgttttccca
     2161 ggaatcgcgg tggtgagtaa ccacgcatca tcaggagtac ggataaaatg cttgatggtc
     2221 gggagaggca taaactccgt cagccagttg agacggacca tctcatctgt aacatcattg
     2281 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaag
     2341 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     2401 tcagcgtcca tgttggagtt taagcgcgga cgggagcaag acgtttcccg ttgaatatgg
     2461 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat
     2521 atatttttat cttgtgcaat gtaacatcag agattttgag acacaacgtg gctttgttga
     2581 ataaatcgaa cttttgctga gttgaaggat cagctctagt agttacattg tcgatctgtt
     2641 catggtgaac agctttaaat gcaccaaaaa ctcgtaaaag ctctgatgta tctatctttt
     2701 ttacaccgtt ttcatctgtg catatggaca gttttccctt tgatatctaa cggtgaacag
     2761 ttgttctact tttgtttgtt agtcttgatg cttcactgat agatacaaga gccataagaa
     2821 cctcagatcc ttccgtattt agccagtatg ttctctagtg tggttcgttg tttttgcgtg
     2881 agccatgaga acgaaccatt gagatcatgc ttactttgca tgtcactcaa aaattttgcc
     2941 tcaaaactgg tgagctgaat ttttgcagtt aaagcatcgt gtagtgtttt tcttagtccg
     3001 ttacgtaggt aggaatctga tgtaatggtt gttggtattt tgtcaccatt catttttatc
     3061 tggttgttct caagttcggt tacgagatcc atttgtctat ctagttcaac ttggaaaatc
     3121 aacgtatcag tcgggcggcc tcgcttatca accaccaatt tcatattgct gtaagtgttt
     3181 aaatctttac ttattggttt caaaacccat tggttaagcc ttttaaactc atggtagtta
     3241 ttttcaagca ttaacatgaa cttaaattca tcaaggctaa tctctatatt tgccttgtga
     3301 gttttctttt gtgttagttc ttttaataac cactcataaa tcctcataga gtatttgttt
     3361 tcaaaagact taacatgttc cagattatat tttatgaatt tttttaactg gaaaagataa
     3421 ggcaatatct cttcactaaa aactaattct aatttttcgc ttgagaactt ggcatagttt
     3481 gtccactgga aaatctcaaa gcctttaacc aaaggattcc tgatttccac agttctcgtc
     3541 atcagctctc tggttgcttt agctaataca ccataagcat tttccctact gatgttcatc
     3601 atctgagcgt attggttata agtgaacgat accgtccgtt ctttccttgt agggttttca
     3661 atcgtggggt tgagtagtgc cacacagcat aaaattagct tggtttcatg ctccgttaag
     3721 tcatagcgac taatcgctag ttcatttgct ttgaaaacaa ctaattcaga catacatctc
     3781 aattggtcta ggtgatttta atcactatac caattgagat gggctagtca atgataatta
     3841 catgtccttt tcctttgagt tgtgggtatc tgtaaattct gctagacctt tgctggaaaa
     3901 cttgtaaatt ctgctagacc ctctgtaaat tccgctagac ctttgtgtgt tttttttgtt
     3961 tatattcaag tggttataat ttatagaata aagaaagaat aaaaaaagat aaaaagaata
     4021 gatcccagcc ctgtgtataa ctcactactt tagtcagttc cgcagtatta caaaaggatg
     4081 tcgcaaacgc tgtttgctcc tctacaaaac agaccttaaa accctaaagg cttaagtagc
     4141 accctcgcaa gctcgggcaa atcgctgaat attccttttg tctccgacca tcaggcacct
     4201 gagtcgctgt ctttttcgtg acattcagtt cgctgcgctc acggctctgg cagtgaatgg
     4261 gggtaaatgg cactacaggc gccttttatg gattcatgca aggaaactac ccataataca
     4321 agaaaagccc gtcacgggct tctcagggcg ttttatggcg ggtctgctat gtggtgctat
     4381 ctgacttttt gctgttcagc agttcctgcc ctctgatttt ccagtctgac cacttcggat
     4441 tatcccgtga caggtcattc agactggcta atgcacccag taaggcagcg gtatcatcaa
     4501 caggcttacc cgtcttactg tccctagtgc ttggattctc accaataaaa aacgcccggc
     4561 ggcaaccgag cgttctgaac aaatccagat ggagttctga ggtcattact ggatctatca
     4621 acaggagtcc aagcgagctc gtaaacttgg tctgacagct ctagctccgg caaaaaaacg
     4681 ggcaaggtgt caccaccctg ccctttttct ttaaaaccga aaagattact tcgcgtttgc
     4741 cacctgacgt ctaagaaaag gaatattcag caatttgccc gtgccgaaga aaggcccacc
     4801 cgtgaaggtg agccagtgag ttgattgcta cgtaattagt tagttagccc ttagtgactc
//
