LOCUS       sequence-13976-e        3163 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 3163)
  AUTHORS   Sleight SC, Bartley BA, Lieviant JA, Sauro HM
  TITLE     Designing and engineering evolutionary robust genetic circuits
  JOURNAL   J Biol Eng. 2010 Nov 1;4:12
  PUBMED    21040586
REFERENCE   2  (bases 1 to 3163)
  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: "J Biol 
            Eng."; date: "2010-11-1"; volume: "4"; pages: "1"
COMMENT     Sequence Label: SS13 lac-inducible GFP reporter circuit
FEATURES             Location/Qualifiers
     source          1..3163
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..21
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(27..44)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(198..786)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(278..297)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(957..1748,1749..1817))
                     /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     1580..1599
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1818..1922)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1990..2008
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2012..2055)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2058..2079
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    2180..2201
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2216..2221,2222..2239,2240..2246)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2254..2270
                     /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             2307..3020
                     /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="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2337..2365)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2960..2980
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      3043..3114
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3130..3157
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 tactagtagc ggccgctgca ggcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
       61 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
      121 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
      181 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
      241 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
      301 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
      361 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
      421 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
      481 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
      541 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
      601 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
      661 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
      721 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
      781 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
      841 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
      901 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
      961 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     1021 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     1081 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     1141 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     1201 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     1261 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     1321 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     1381 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     1441 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     1501 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     1561 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     1621 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     1681 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     1741 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     1801 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     1861 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     1921 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     1981 acctataaaa ataggcgtat cacgaggcag aatttcagat aaaaaaaatc cttagctttc
     2041 gctaaggatg atttctggaa ttcgcggccg cttctagagc aatacgcaaa ccgcctctcc
     2101 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     2161 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     2221 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacat
     2281 actagagtca cacaggaaag tactagatgc gtaaaggaga agaacttttc actggagttg
     2341 tcccaattct tgttgaatta gatggtgatg ttaatgggca caaattttct gtcagtggag
     2401 agggtgaagg tgatgcaaca tacggaaaac ttacccttaa atttatttgc actactggaa
     2461 aactacctgt tccatggcca acacttgtca ctactttcgg ttatggtgtt caatgctttg
     2521 cgagataccc agatcatatg aaacagcatg actttttcaa gagtgccatg cccgaaggtt
     2581 atgtacagga aagaactata tttttcaaag atgacgggaa ctacaagaca cgtgctgaag
     2641 tcaagtttga aggtgatacc cttgttaata gaatcgagtt aaaaggtatt gattttaaag
     2701 aagatggaaa cattcttgga cacaaattgg aatacaacta taactcacac aatgtataca
     2761 tcatggcaga caaacaaaag aatggaatca aagttaactt caaaattaga cacaacattg
     2821 aagatggaag cgttcaacta gcagaccatt atcaacaaaa tactccaatt ggcgatggcc
     2881 ctgtcctttt accagacaac cattacctgt ccacacaatc tgccctttcg aaagatccca
     2941 acgaaaagag agaccacatg gtccttcttg agtttgtaac agctgctggg attacacatg
     3001 gcatggatga actatacaaa taataatact agagccaggc atcaaataaa acgaaaggct
     3061 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag
     3121 agtcacactg gctcaccttc gggtgggcct ttctgcgttt ata
//
