LOCUS       sequence-49411-e        4143 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 4143)
  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 4143)
  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: pSTC0r1
FEATURES             Location/Qualifiers
     source          1..4143
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    2..23
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    62..78
                     /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)."
     CDS             127..834
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="KGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFI
                     CTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNY
                     KTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNF
                     KIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFV
                     TAAGITHGMDELYK"
     primer_bind     complement(151..179)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     774..794
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      857..928
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      944..971
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    978..998
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(1082..1099)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1253..1841)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1333..1352)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2074..2889)
                     /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"
     CDS             complement(2074..2889)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /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     2797..2816
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2881..2900
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(join(3044..3835,3836..3904))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3667..3686
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3905..4009)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4077..4095
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(4099..4142)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 ggaattcgcg gccgcttcta gagatgcatg ctagcataaa tgtgagcgga taacattgac
       61 attgtgagcg gataacaaga tactgagcac aactagagaa agaggaaaaa ctagatgact
      121 agccgtaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt
      181 gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc aacatacgga
      241 aaacttaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg gccaacactt
      301 gtcactactt tcggttatgg tgttcaatgc tttgcgagat acccagatca tatgaaacag
      361 catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaagaac tatatttttc
      421 aaagatgacg ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga tacccttgtt
      481 aatagaatcg agttaaaagg tattgatttt aaagaagatg gaaacattct tggacacaaa
      541 ttggaataca actataactc acacaatgta tacatcatgg cagacaaaca aaagaatgga
      601 atcaaagtta acttcaaaat tagacacaac attgaagatg gaagcgttca actagcagac
      661 cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac
      721 ctgtccacac aatctgccct ttcgaaagat cccaacgaaa agagagacca catggtcctt
      781 cttgagtttg taacagctgc tgggattaca catggcatgg atgaactata caaataataa
      841 tactagagcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
      901 tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg
      961 gcctttctgc gtttatatac tagtagcggc cgctgcagtc cggcaaaaaa acgggcaagg
     1021 tgtcaccacc ctgccctttt tctttaaaac cgaaaagatt acttcgcgtt atgcaggctt
     1081 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     1141 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     1201 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     1261 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     1321 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     1381 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     1441 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     1501 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     1561 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     1621 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     1681 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     1741 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     1801 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     1861 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     1921 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     1981 aaagtatata tgagtaaact tggtctgaca gctcgagtcc cgtcaagtca gcgtaatgct
     2041 ctgccagtgt tacaaccaat taaccaattc tgattagaaa aactcatcga gcatcaaatg
     2101 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
     2161 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     2221 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     2281 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagctt
     2341 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     2401 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc
     2461 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     2521 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     2581 cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     2641 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     2701 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     2761 caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     2821 taaatcagca tccatgttgg aatttaatcg cggcctggag caagacgttt cccgttgaat
     2881 atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga
     2941 tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacaa cgtggctttg
     3001 ttgaataaat cgaacttttg ctgagttgaa ggatcagctc gagttaccaa tgcttaatca
     3061 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3121 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3181 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     3241 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     3301 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     3361 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     3421 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     3481 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     3541 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     3601 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     3661 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     3721 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     3781 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     3841 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     3901 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     3961 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4021 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     4081 ggcgtatcac gaggcagaat ttcagataaa aaaaatcctt agctttcgct aaggatgatt
     4141 tct
//
