LOCUS       sequence-60654-e        4923 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 4923)
  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 4923)
  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: XNOR-gate-V2.0
FEATURES             Location/Qualifiers
     source          1..4923
                     /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      complement(185..212)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(228..299)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    309..356
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     misc_RNA        413..462
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             556..564
                     /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             571..1284
                     /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
                     DFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1307..1378
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1394..1421
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1428..1448
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1484..1515
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     terminator      1640..1734
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2226..2448
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             2496..3446
                     /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(3735..3803,3804..4595)
                     /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(4725..4754)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     terminator      complement(4848..4877)
                     /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 atccgggcta
      181 taaacgcaga aaggcccacc cgaaggtgag ccagtgtgac tctagtagag agcgttcacc
      241 gacaaacaac agataaaacg aaaggcccag tctttcgact gagcctttcg ttttatttga
      301 tgcctggggg tttgtaccgt acaccactga gaccgcggtg gttgaccaga caaaccacga
      361 aaaggagttt tttagttacc ttaattgaaa taaacgaaat aaaaactcgc agctgtcgcc
      421 ggatgtgctt tccggtctga tgagtccgtg aggacgaaac agcctctaca aataattttg
      481 tttaagggcc caagttcact taaaaaggag atcaacaatg aaagcaattt tcgtactgaa
      541 acatcttaat catgctaagg aggttttcta atgcgtaaag gcgaagagct gttcactggt
      601 gtcgtcccta ttctggtgga actggatggt gatgtcaacg gtcataagtt ttccgtgcgt
      661 ggcgagggtg aaggtgacgc aactaatggt aaactgacgc tgaagttcat ctgtactact
      721 ggtaaactgc cggtaccttg gccgactctg gtaacgacgc tgacttatgg tgttcagtgc
      781 tttgctcgtt atccggacca tatgaagcag catgacttct tcaagtccgc catgccggaa
      841 ggctatgtgc aggaacgcac gatttccttt aaggatgacg gcacgtacaa aacgcgtgcg
      901 gaagtgaaat ttgaaggcga tactctggta aaccgcattg agctgaaagg cattgacttt
      961 aaagaagacg gcaatatcct gggccataag ctggaataca attttaacag ccacaatgtt
     1021 tacatcaccg ccgataaaca aaaaaatggc actaaagcgg attttaaaat tcgccacaac
     1081 gtggaggatg gcagcgtgca gctggctgat cactaccagc aaaacactcc aatcggtgat
     1141 ggtcctgttc tgctgccaga caatcactat ctgagcacgc aaagcgttct gtctaaagat
     1201 ccgaacgaga aacgcgatca tatggttctg ctggagttcg taaccgcagc gggcatcacg
     1261 catggtatgg atgaactgta caaatgatga tactagagcc aggcatcaaa taaaacgaaa
     1321 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcta
     1381 ctagagtcac actggctcac cttcgggtgg gcctttctgc gtttatatac tagtagcggc
     1441 cgctgcagga gtcactaagg gttagttagt tagattagca gaaagtcaaa agcctccgac
     1501 cggaggcttt tgactaaaac ttcccttggg gttatcattg gggctcactc aaaggcggta
     1561 atcagataaa aaaaatcctt agctttcgct aaggatgatt tctgctagag ctgtcagacc
     1621 aagtttacga gctcgcttgg actcctgttg atagatccag taatgacctc agaactccat
     1681 ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtga gaatccaagc
     1741 actagggaca gtaagacggg taagcctgtt gatgataccg ctgccttact gggtgcatta
     1801 gccagtctga atgacctgtc acgggataat ccgaagtggt cagactggaa aatcagaggg
     1861 caggaactgc tgaacagcaa aaagtcagat agcaccacat agcagacccg ccataaaacg
     1921 ccctgagaag cccgtgacgg gcttttcttg tattatgggt agtttccttg catgaatcca
     1981 taaaaggcgc ctgtagtgcc atttaccccc attcactgcc agagccgtga gcgcagcgaa
     2041 ctgaatgtca cgaaaaagac agcgactcag gtgcctgatg gtcggagaca aaaggaatat
     2101 tcagcgattt gcccgagctt gcgagggtgc tacttaagcc tttagggttt taaggtctgt
     2161 tttgtagagg agcaaacagc gtttgcgaca tccttttgta atactgcgga actgactaaa
     2221 gtagtgagtt atacacaggg ctgggatcta ttctttttat ctttttttat tctttcttta
     2281 ttctataaat tataaccact tgaatataaa caaaaaaaac acacaaaggt ctagcggaat
     2341 ttacagaggg tctagcagaa tttacaagtt ttccagcaaa ggtctagcag aatttacaga
     2401 tacccacaac tcaaaggaaa aggacatgta attatcattg actagcccat ctcaattggt
     2461 atagtgatta aaatcaccta gaccaattga gatgtatgtc tgaattagtt gttttcaaag
     2521 caaatgaact agcgattagt cgctatgact taacggagca tgaaaccaag ctaattttat
     2581 gctgtgtggc actactcaac cccacgattg aaaaccctac aaggaaagaa cggacggtat
     2641 cgttcactta taaccaatac gctcagatga tgaacatcag tagggaaaat gcttatggtg
     2701 tattagctaa agcaaccaga gagctgatga cgagaactgt ggaaatcagg aatcctttgg
     2761 ttaaaggctt tgagattttc cagtggacaa actatgccaa gttctcaagc gaaaaattag
     2821 aattagtttt tagtgaagag atattgcctt atcttttcca gttaaaaaaa ttcataaaat
     2881 ataatctgga acatgttaag tcttttgaaa acaaatactc tatgaggatt tatgagtggt
     2941 tattaaaaga actaacacaa aagaaaactc acaaggcaaa tatagagatt agccttgatg
     3001 aatttaagtt catgttaatg cttgaaaata actaccatga gtttaaaagg cttaaccaat
     3061 gggttttgaa accaataagt aaagatttaa acacttacag caatatgaaa ttggtggttg
     3121 ataagcgagg ccgcccgact gatacgttga ttttccaagt tgaactagat agacaaatgg
     3181 atctcgtaac cgaacttgag aacaaccaga taaaaatgaa tggtgacaaa ataccaacaa
     3241 ccattacatc agattcctac ctacgtaacg gactaagaaa aacactacac gatgctttaa
     3301 ctgcaaaaat tcagctcacc agttttgagg caaaattttt gagtgacatg caaagtaagc
     3361 atgatctcaa tggttcgttc tcatggctca cgcaaaaaca acgaaccaca ctagagaaca
     3421 tactggctaa atacggaagg atctgaggtt cttatggctc ttgtatctat cagtgaagca
     3481 tcaagactaa caaacaaaag tagaacaact gttcaccgtt agatatcaaa gggaaaactg
     3541 tccatatgca cagatgaaaa cggtgtaaaa aagatagata catcagagct tttacgagtt
     3601 tttggtgcat ttaaagctgt tcaccatgaa cagatcgaca atgtaactac tagagctaaa
     3661 tacattcaaa tatctatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3721 gaaaaaggaa gaatatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3781 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgccgaag
     3841 atcagttggg tgcacgtgtg ggttacatcg aactggacct caacagcggt aagattcttg
     3901 agagttttcg ccccgaagaa cgtttcccaa tgatgagcac ttttaaagtt ctgctctgtg
     3961 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4021 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gacggcatga
     4081 cagtacgcga attatgcagc gctgccataa ccatgagtga taacacggcg gccaacttac
     4141 ttctgacaac gatcggagga ccgaaggagc ttaccgcttt tttgcacaac atgggtgatc
     4201 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4261 gtgacaccac gatgcctgta gctatggcaa caacgttgcg caaactctta actggcgaac
     4321 ttcttactct cgcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4381 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4441 gtgagcgtgg gtcccgcggt attattgcag ccctggggcc agatggtaag ccctcccgta
     4501 tcgtagttat ctacacgacg gggagccagg caactatgga cgaacgtaat cgccagatcg
     4561 ctgagatagg tgcctccctg attaagcatt ggtaataata ctagctccgg caaaaaaacg
     4621 ggcaaggtgt caccaccctg ccctttttct ttaaaaccga aaagattact tcgcgtttgc
     4681 cacctgacgt ctaagaaaag gaatattcag caatttgccc gtgccgaaga aaggcccacc
     4741 cgtgaaggtg agccagtgag ttgattgcta cgtaattagt tagttagccc ttagtgactc
     4801 tgccacctga cgtctaagaa aaggaatatt cagcaatttg cccgtgccga agaaaggccc
     4861 acccgtgaag gtgagccagt gagttgattg ctacgtaatt agttagttag cccttagtga
     4921 ctc
//
