LOCUS       sequence-60655-e        4822 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 4822)
  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 4822)
  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-ASV
FEATURES             Location/Qualifiers
     source          1..4822
                     /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
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     misc_feature    1327..1347
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1383..1414
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     terminator      1539..1633
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2125..2347
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             2395..3345
                     /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(3634..3702,3703..4494)
                     /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(4624..4653)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     terminator      complement(4747..4776)
                     /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 agctgtcacc
      421 ggatgtgctt tccggcctga 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 actaaagcga 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 caaagccggc gctgctaacg acgaaaacta cgctgcgtct
     1321 gtttaatact agtagcggcc gctgcaggag tcactaaggg ttagttagtt agattagcag
     1381 aaagtcaaaa gcctccgacc ggaggctttt gactaaaact tcccttgggg ttatcattgg
     1441 ggctcactca aaggcggtaa tcagataaaa aaaatcctta gctttcgcta aggatgattt
     1501 ctgctagagc tgtcagacca agtttacgag ctcgcttgga ctcctgttga tagatccagt
     1561 aatgacctca gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt
     1621 tattggtgag aatccaagca ctagggacag taagacgggt aagcctgttg atgataccgc
     1681 tgccttactg ggtgcattag ccagtctgaa tgacctgtca cgggataatc cgaagtggtc
     1741 agactggaaa atcagagggc aggaactgct gaacagcaaa aagtcagata gcaccacata
     1801 gcagacccgc cataaaacgc cctgagaagc ccgtgacggg cttttcttgt attatgggta
     1861 gtttccttgc atgaatccat aaaaggcgcc tgtagtgcca tttaccccca ttcactgcca
     1921 gagccgtgag cgcagcgaac tgaatgtcac gaaaaagaca gcgactcagg tgcctgatgg
     1981 tcggagacaa aaggaatatt cagcgatttg cccgagcttg cgagggtgct acttaagcct
     2041 ttagggtttt aaggtctgtt ttgtagagga gcaaacagcg tttgcgacat ccttttgtaa
     2101 tactgcggaa ctgactaaag tagtgagtta tacacagggc tgggatctat tctttttatc
     2161 tttttttatt ctttctttat tctataaatt ataaccactt gaatataaac aaaaaaaaca
     2221 cacaaaggtc tagcggaatt tacagagggt ctagcagaat ttacaagttt tccagcaaag
     2281 gtctagcaga atttacagat acccacaact caaaggaaaa ggacatgtaa ttatcattga
     2341 ctagcccatc tcaattggta tagtgattaa aatcacctag accaattgag atgtatgtct
     2401 gaattagttg ttttcaaagc aaatgaacta gcgattagtc gctatgactt aacggagcat
     2461 gaaaccaagc taattttatg ctgtgtggca ctactcaacc ccacgattga aaaccctaca
     2521 aggaaagaac ggacggtatc gttcacttat aaccaatacg ctcagatgat gaacatcagt
     2581 agggaaaatg cttatggtgt attagctaaa gcaaccagag agctgatgac gagaactgtg
     2641 gaaatcagga atcctttggt taaaggcttt gagattttcc agtggacaaa ctatgccaag
     2701 ttctcaagcg aaaaattaga attagttttt agtgaagaga tattgcctta tcttttccag
     2761 ttaaaaaaat tcataaaata taatctggaa catgttaagt cttttgaaaa caaatactct
     2821 atgaggattt atgagtggtt attaaaagaa ctaacacaaa agaaaactca caaggcaaat
     2881 atagagatta gccttgatga atttaagttc atgttaatgc ttgaaaataa ctaccatgag
     2941 tttaaaaggc ttaaccaatg ggttttgaaa ccaataagta aagatttaaa cacttacagc
     3001 aatatgaaat tggtggttga taagcgaggc cgcccgactg atacgttgat tttccaagtt
     3061 gaactagata gacaaatgga tctcgtaacc gaacttgaga acaaccagat aaaaatgaat
     3121 ggtgacaaaa taccaacaac cattacatca gattcctacc tacgtaacgg actaagaaaa
     3181 acactacacg atgctttaac tgcaaaaatt cagctcacca gttttgaggc aaaatttttg
     3241 agtgacatgc aaagtaagca tgatctcaat ggttcgttct catggctcac gcaaaaacaa
     3301 cgaaccacac tagagaacat actggctaaa tacggaagga tctgaggttc ttatggctct
     3361 tgtatctatc agtgaagcat caagactaac aaacaaaagt agaacaactg ttcaccgtta
     3421 gatatcaaag ggaaaactgt ccatatgcac agatgaaaac ggtgtaaaaa agatagatac
     3481 atcagagctt ttacgagttt ttggtgcatt taaagctgtt caccatgaac agatcgacaa
     3541 tgtaactact agagctaaat acattcaaat atctatccgc tcatgagaca ataaccctga
     3601 taaatgcttc aataatattg aaaaaggaag aatatgagta ttcaacattt ccgtgtcgcc
     3661 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     3721 aaagtaaaag atgccgaaga tcagttgggt gcacgtgtgg gttacatcga actggacctc
     3781 aacagcggta agattcttga gagttttcgc cccgaagaac gtttcccaat gatgagcact
     3841 tttaaagttc tgctctgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     3901 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     3961 catcttacgg acggcatgac agtacgcgaa ttatgcagcg ctgccataac catgagtgat
     4021 aacacggcgg ccaacttact tctgacaacg atcggaggac cgaaggagct taccgctttt
     4081 ttgcacaaca tgggtgatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     4141 gccataccaa acgacgagcg tgacaccacg atgcctgtag ctatggcaac aacgttgcgc
     4201 aaactcttaa ctggcgaact tcttactctc gcttcccggc aacaattaat agactggatg
     4261 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     4321 gctgataaat ctggagccgg tgagcgtggg tcccgcggta ttattgcagc cctggggcca
     4381 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagccaggc aactatggac
     4441 gaacgtaatc gccagatcgc tgagataggt gcctccctga ttaagcattg gtaataatac
     4501 tagctccggc aaaaaaacgg gcaaggtgtc accaccctgc cctttttctt taaaaccgaa
     4561 aagattactt cgcgtttgcc acctgacgtc taagaaaagg aatattcagc aatttgcccg
     4621 tgccgaagaa aggcccaccc gtgaaggtga gccagtgagt tgattgctac gtaattagtt
     4681 agttagccct tagtgactct gccacctgac gtctaagaaa aggaatattc agcaatttgc
     4741 ccgtgccgaa gaaaggccca cccgtgaagg tgagccagtg agttgattgc tacgtaatta
     4801 gttagttagc ccttagtgac tc
//
