LOCUS       sequence-60653-e        4935 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 4935)
  AUTHORS   Bonnet J, Yin P, Ortiz ME, Subsoontorn P, Endy D
  TITLE     Amplifying genetic logic gates.
  JOURNAL   Science. 2013 May 3;340(6132):599-603. doi: 10.1126/science.1232758.
            Epub 2013 Mar 28.
  PUBMED    23539178
REFERENCE   2  (bases 1 to 4935)
  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: 
            "10.1126/science.1232758"; journalName: "Science"; date: "2013-05-3-
            3"; volume: "340"; issue: "6132"; pages: "599-603"
COMMENT     Sequence Label: XOR gate-V2.0
FEATURES             Location/Qualifiers
     source          1..4935
                     /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    129..178
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     terminator      193..264
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      280..307
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    321..368
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     misc_RNA        425..474
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             568..576
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             583..1296
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGTKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1319..1390
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1406..1433
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1440..1460
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1496..1527
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     terminator      1652..1746
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2238..2460
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             2508..3458
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     CDS             join(3747..3815,3816..4607)
                     /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"
     terminator      complement(4737..4766)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     terminator      complement(4860..4889)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 gaattcgcgg ccgcttctag agaaaaaatt tatttgcttt cgcatctttt tgtacctata
       61 atgtgtggaa agcttatgcc aacacaatta acatctcaat caaggtaaat gctttttgct
      121 ttttttgctc ggccggcttg tcgacgacgg cggtctccgt cgtcaggatc atccgggcca
      181 tatgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
      241 tgttgtttgt cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct
      301 ttctgcgttt ataatgcatg ggtttgtacc gtacaccact gagaccgcgg tggttgacca
      361 gacaaaccac gaaaaggagt tttttagtta ccttaattga aataaacgaa ataaaaactc
      421 gcagctgtca ccggatgtgc tttccggtct gatgagtccg tgaggacgaa acagcctcta
      481 caaataattt tgtttaaggg cccaagttca cttaaaaagg agatcaacaa tgaaagcaat
      541 tttcgtactg aaacatctta atcatgctaa ggaggttttc taatgcgtaa aggcgaagag
      601 ctgttcactg gtgtcgtccc tattctggtg gaactggatg gtgatgtcaa cggtcataag
      661 ttttccgtgc gtggcgaggg tgaaggtgac gcaactaatg gtaaactgac gctgaagttc
      721 atctgtacta ctggtaaact gccggtacct tggccgactc tggtaacgac gctgacttat
      781 ggtgttcagt gctttgctcg ttatccggac catatgaagc agcatgactt cttcaagtcc
      841 gccatgccgg aaggctatgt gcaggaacgc acgatttcct ttaaggatga cggcacgtac
      901 aaaacgcgtg cggaagtgaa atttgaaggc gatactctgg taaaccgcat tgagctgaaa
      961 ggcattgact ttaaagaaga cggcaatatc ctgggccata agctggaata caattttaac
     1021 agccacaatg tttacatcac cgccgataaa caaaaaaatg gcactaaagc gaattttaaa
     1081 attcgccaca acgtggagga tggcagcgtg cagctggctg atcactacca gcaaaacact
     1141 ccaatcggtg atggtcctgt tctgctgcca gacaatcact atctgagcac gcaaagcgtt
     1201 ctgtctaaag atccgaacga gaaacgcgat catatggttc tgctggagtt cgtaaccgca
     1261 gcgggcatca cgcatggtat ggatgaactg tacaaatgat gatactagag ccaggcatca
     1321 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     1381 gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct gcgtttatat
     1441 actagtagcg gccgctgcag gagtcactaa gggttagtta gttagattag cagaaagtca
     1501 aaagcctccg accggaggct tttgactaaa acttcccttg gggttatcat tggggctcac
     1561 tcaaaggcgg taatcagata aaaaaaatcc ttagctttcg ctaaggatga tttctgctag
     1621 agctgtcaga ccaagtttac gagctcgctt ggactcctgt tgatagatcc agtaatgacc
     1681 tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt
     1741 gagaatccaa gcactaggga cagtaagacg ggtaagcctg ttgatgatac cgctgcctta
     1801 ctgggtgcat tagccagtct gaatgacctg tcacgggata atccgaagtg gtcagactgg
     1861 aaaatcagag ggcaggaact gctgaacagc aaaaagtcag atagcaccac atagcagacc
     1921 cgccataaaa cgccctgaga agcccgtgac gggcttttct tgtattatgg gtagtttcct
     1981 tgcatgaatc cataaaaggc gcctgtagtg ccatttaccc ccattcactg ccagagccgt
     2041 gagcgcagcg aactgaatgt cacgaaaaag acagcgactc aggtgcctga tggtcggaga
     2101 caaaaggaat attcagcgat ttgcccgagc ttgcgagggt gctacttaag cctttagggt
     2161 tttaaggtct gttttgtaga ggagcaaaca gcgtttgcga catccttttg taatactgcg
     2221 gaactgacta aagtagtgag ttatacacag ggctgggatc tattcttttt atcttttttt
     2281 attctttctt tattctataa attataacca cttgaatata aacaaaaaaa acacacaaag
     2341 gtctagcgga atttacagag ggtctagcag aatttacaag ttttccagca aaggtctagc
     2401 agaatttaca gatacccaca actcaaagga aaaggacatg taattatcat tgactagccc
     2461 atctcaattg gtatagtgat taaaatcacc tagaccaatt gagatgtatg tctgaattag
     2521 ttgttttcaa agcaaatgaa ctagcgatta gtcgctatga cttaacggag catgaaacca
     2581 agctaatttt atgctgtgtg gcactactca accccacgat tgaaaaccct acaaggaaag
     2641 aacggacggt atcgttcact tataaccaat acgctcagat gatgaacatc agtagggaaa
     2701 atgcttatgg tgtattagct aaagcaacca gagagctgat gacgagaact gtggaaatca
     2761 ggaatccttt ggttaaaggc tttgagattt tccagtggac aaactatgcc aagttctcaa
     2821 gcgaaaaatt agaattagtt tttagtgaag agatattgcc ttatcttttc cagttaaaaa
     2881 aattcataaa atataatctg gaacatgtta agtcttttga aaacaaatac tctatgagga
     2941 tttatgagtg gttattaaaa gaactaacac aaaagaaaac tcacaaggca aatatagaga
     3001 ttagccttga tgaatttaag ttcatgttaa tgcttgaaaa taactaccat gagtttaaaa
     3061 ggcttaacca atgggttttg aaaccaataa gtaaagattt aaacacttac agcaatatga
     3121 aattggtggt tgataagcga ggccgcccga ctgatacgtt gattttccaa gttgaactag
     3181 atagacaaat ggatctcgta accgaacttg agaacaacca gataaaaatg aatggtgaca
     3241 aaataccaac aaccattaca tcagattcct acctacgtaa cggactaaga aaaacactac
     3301 acgatgcttt aactgcaaaa attcagctca ccagttttga ggcaaaattt ttgagtgaca
     3361 tgcaaagtaa gcatgatctc aatggttcgt tctcatggct cacgcaaaaa caacgaacca
     3421 cactagagaa catactggct aaatacggaa ggatctgagg ttcttatggc tcttgtatct
     3481 atcagtgaag catcaagact aacaaacaaa agtagaacaa ctgttcaccg ttagatatca
     3541 aagggaaaac tgtccatatg cacagatgaa aacggtgtaa aaaagataga tacatcagag
     3601 cttttacgag tttttggtgc atttaaagct gttcaccatg aacagatcga caatgtaact
     3661 actagagcta aatacattca aatatctatc cgctcatgag acaataaccc tgataaatgc
     3721 ttcaataata ttgaaaaagg aagaatatga gtattcaaca tttccgtgtc gcccttattc
     3781 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3841 aagatgccga agatcagttg ggtgcacgtg tgggttacat cgaactggac ctcaacagcg
     3901 gtaagattct tgagagtttt cgccccgaag aacgtttccc aatgatgagc acttttaaag
     3961 ttctgctctg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     4021 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     4081 cggacggcat gacagtacgc gaattatgca gcgctgccat aaccatgagt gataacacgg
     4141 cggccaactt acttctgaca acgatcggag gaccgaagga gcttaccgct tttttgcaca
     4201 acatgggtga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     4261 caaacgacga gcgtgacacc acgatgcctg tagctatggc aacaacgttg cgcaaactct
     4321 taactggcga acttcttact ctcgcttccc ggcaacaatt aatagactgg atggaggcgg
     4381 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     4441 aatctggagc cggtgagcgt gggtcccgcg gtattattgc agccctgggg ccagatggta
     4501 agccctcccg tatcgtagtt atctacacga cggggagcca ggcaactatg gacgaacgta
     4561 atcgccagat cgctgagata ggtgcctccc tgattaagca ttggtaataa tactagctcc
     4621 ggcaaaaaaa cgggcaaggt gtcaccaccc tgcccttttt ctttaaaacc gaaaagatta
     4681 cttcgcgttt gccacctgac gtctaagaaa aggaatattc agcaatttgc ccgtgccgaa
     4741 gaaaggccca cccgtgaagg tgagccagtg agttgattgc tacgtaatta gttagttagc
     4801 ccttagtgac tctgccacct gacgtctaag aaaaggaata ttcagcaatt tgcccgtgcc
     4861 gaagaaaggc ccacccgtga aggtgagcca gtgagttgat tgctacgtaa ttagttagtt
     4921 agcccttagt gactc
//
