LOCUS       sequence-49409-e        4042 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 4042)
  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 4042)
  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: pSTC0
FEATURES             Location/Qualifiers
     source          1..4042
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             26..733
                     /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(50..78)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     673..693
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      756..827
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      843..870
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    877..897
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(981..998)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1152..1740)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1232..1251)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1973..2788)
                     /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(1973..2788)
                     /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     2696..2715
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2780..2799
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(join(2943..3734,3735..3803))
                     /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     3566..3585
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3804..3908)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3976..3994
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3998..4041)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 ggaattcgcg gccgctgcta gccgtaaagg agaagaactt ttcactggag ttgtcccaat
       61 tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga
      121 aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc
      181 tgttccatgg ccaacacttg tcactacttt cggttatggt gttcaatgct ttgcgagata
      241 cccagatcat atgaaacagc atgacttttt caagagtgcc atgcccgaag gttatgtaca
      301 ggaaagaact atatttttca aagatgacgg gaactacaag acacgtgctg aagtcaagtt
      361 tgaaggtgat acccttgtta atagaatcga gttaaaaggt attgatttta aagaagatgg
      421 aaacattctt ggacacaaat tggaatacaa ctataactca cacaatgtat acatcatggc
      481 agacaaacaa aagaatggaa tcaaagttaa cttcaaaatt agacacaaca ttgaagatgg
      541 aagcgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct
      601 tttaccagac aaccattacc tgtccacaca atctgccctt tcgaaagatc ccaacgaaaa
      661 gagagaccac atggtccttc ttgagtttgt aacagctgct gggattacac atggcatgga
      721 tgaactatac aaataataat actagagcca ggcatcaaat aaaacgaaag gctcagtcga
      781 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca
      841 ctggctcacc ttcgggtggg cctttctgcg tttatatact agtagcggcc gctgcagtcc
      901 ggcaaaaaaa cgggcaaggt gtcaccaccc tgcccttttt ctttaaaacc gaaaagatta
      961 cttcgcgtta tgcaggcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     1021 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     1081 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     1141 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     1201 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     1261 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     1321 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     1381 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     1441 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     1501 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     1561 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     1621 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     1681 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     1741 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     1801 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     1861 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ctcgagtccc
     1921 gtcaagtcag cgtaatgctc tgccagtgtt acaaccaatt aaccaattct gattagaaaa
     1981 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt
     2041 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     2101 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     2161 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
     2221 gtgagaatgg caaaagctta tgcatttctt tccagacttg ttcaacaggc cagccattac
     2281 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
     2341 cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
     2401 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
     2461 atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag
     2521 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
     2581 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
     2641 gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc
     2701 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctggagc
     2761 aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag
     2821 acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt
     2881 gagacacaac gtggctttgt tgaataaatc gaacttttgc tgagttgaag gatcagctcg
     2941 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     3001 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     3061 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     3121 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     3181 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     3241 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     3301 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     3361 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     3421 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     3481 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     3541 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     3601 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     3661 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     3721 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     3781 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     3841 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     3901 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     3961 acattaacct ataaaaatag gcgtatcacg aggcagaatt tcagataaaa aaaatcctta
     4021 gctttcgcta aggatgattt ct
//
