LOCUS       sequence-60648-e        5042 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 5042)
  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 5042)
  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: AND-gate-V2.0
FEATURES             Location/Qualifiers
     source          1..5042
                     /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"""
     terminator      137..208
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      224..251
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    314..363
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     protein_bind    478..525
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     misc_RNA        532..581
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             675..683
                     /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             690..1403
                     /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      1426..1497
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1513..1540
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1547..1567
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1603..1634
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     terminator      1759..1853
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2345..2567
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             2615..3565
                     /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(3854..3922,3923..4714)
                     /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(4844..4873)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     terminator      complement(4967..4996)
                     /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 ttttttgccc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
      181 tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg
      241 gcctttctgc gtttataaaa ggagtttttt agttacctta attgaaataa acgaaataaa
      301 aactcgcgct agctcggccg gcttgtcgac gacggcggtc tccgtcgtca ggatcatccg
      361 ggcccggctt atcggtcagt ttcacctgat ttacgtaaaa acccgcttcg gcgggttttt
      421 gcttttggag gggcagaaag atgaatgact gtccacgacg ctatacccaa aagaaagggt
      481 ttgtaccgta caccactgag accgcggtgg ttgaccagac aaaccacgaa gctgtcaccg
      541 gatgtgcttt ccggtctgat gagtccgtga ggacgaaaca gcctctacaa ataattttgt
      601 ttaagggccc aagttcactt aaaaaggaga tcaacaatga aagcaatttt cgtactgaaa
      661 catcttaatc atgctaagga ggttttctaa tgcgtaaagg cgaagagctg ttcactggtg
      721 tcgtccctat tctggtggaa ctggatggtg atgtcaacgg tcataagttt tccgtgcgtg
      781 gcgagggtga aggtgacgca actaatggta aactgacgct gaagttcatc tgtactactg
      841 gtaaactgcc ggtaccttgg ccgactctgg taacgacgct gacttatggt gttcagtgct
      901 ttgctcgtta tccggaccat atgaagcagc atgacttctt caagtccgcc atgccggaag
      961 gctatgtgca ggaacgcacg atttccttta aggatgacgg cacgtacaaa acgcgtgcgg
     1021 aagtgaaatt tgaaggcgat actctggtaa accgcattga gctgaaaggc attgacttta
     1081 aagaagacgg caatatcctg ggccataagc tggaatacaa ttttaacagc cacaatgttt
     1141 acatcaccgc cgataaacaa aaaaatggca ctaaagcgaa ttttaaaatt cgccacaacg
     1201 tggaggatgg cagcgtgcag ctggctgatc actaccagca aaacactcca atcggtgatg
     1261 gtcctgttct gctgccagac aatcactatc tgagcacgca aagcgttctg tctaaagatc
     1321 cgaacgagaa acgcgatcat atggttctgc tggagttcgt aaccgcagcg ggcatcacgc
     1381 atggtatgga tgaactgtac aaatgatgat actagagcca ggcatcaaat aaaacgaaag
     1441 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac
     1501 tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttatatact agtagcggcc
     1561 gctgcaggag tcactaaggg ttagttagtt agattagcag aaagtcaaaa gcctccgacc
     1621 ggaggctttt gactaaaact tcccttgggg ttatcattgg ggctcactca aaggcggtaa
     1681 tcagataaaa aaaatcctta gctttcgcta aggatgattt ctgctagagc tgtcagacca
     1741 agtttacgag ctcgcttgga ctcctgttga tagatccagt aatgacctca gaactccatc
     1801 tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccaagca
     1861 ctagggacag taagacgggt aagcctgttg atgataccgc tgccttactg ggtgcattag
     1921 ccagtctgaa tgacctgtca cgggataatc cgaagtggtc agactggaaa atcagagggc
     1981 aggaactgct gaacagcaaa aagtcagata gcaccacata gcagacccgc cataaaacgc
     2041 cctgagaagc ccgtgacggg cttttcttgt attatgggta gtttccttgc atgaatccat
     2101 aaaaggcgcc tgtagtgcca tttaccccca ttcactgcca gagccgtgag cgcagcgaac
     2161 tgaatgtcac gaaaaagaca gcgactcagg tgcctgatgg tcggagacaa aaggaatatt
     2221 cagcgatttg cccgagcttg cgagggtgct acttaagcct ttagggtttt aaggtctgtt
     2281 ttgtagagga gcaaacagcg tttgcgacat ccttttgtaa tactgcggaa ctgactaaag
     2341 tagtgagtta tacacagggc tgggatctat tctttttatc tttttttatt ctttctttat
     2401 tctataaatt ataaccactt gaatataaac aaaaaaaaca cacaaaggtc tagcggaatt
     2461 tacagagggt ctagcagaat ttacaagttt tccagcaaag gtctagcaga atttacagat
     2521 acccacaact caaaggaaaa ggacatgtaa ttatcattga ctagcccatc tcaattggta
     2581 tagtgattaa aatcacctag accaattgag atgtatgtct gaattagttg ttttcaaagc
     2641 aaatgaacta gcgattagtc gctatgactt aacggagcat gaaaccaagc taattttatg
     2701 ctgtgtggca ctactcaacc ccacgattga aaaccctaca aggaaagaac ggacggtatc
     2761 gttcacttat aaccaatacg ctcagatgat gaacatcagt agggaaaatg cttatggtgt
     2821 attagctaaa gcaaccagag agctgatgac gagaactgtg gaaatcagga atcctttggt
     2881 taaaggcttt gagattttcc agtggacaaa ctatgccaag ttctcaagcg aaaaattaga
     2941 attagttttt agtgaagaga tattgcctta tcttttccag ttaaaaaaat tcataaaata
     3001 taatctggaa catgttaagt cttttgaaaa caaatactct atgaggattt atgagtggtt
     3061 attaaaagaa ctaacacaaa agaaaactca caaggcaaat atagagatta gccttgatga
     3121 atttaagttc atgttaatgc ttgaaaataa ctaccatgag tttaaaaggc ttaaccaatg
     3181 ggttttgaaa ccaataagta aagatttaaa cacttacagc aatatgaaat tggtggttga
     3241 taagcgaggc cgcccgactg atacgttgat tttccaagtt gaactagata gacaaatgga
     3301 tctcgtaacc gaacttgaga acaaccagat aaaaatgaat ggtgacaaaa taccaacaac
     3361 cattacatca gattcctacc tacgtaacgg actaagaaaa acactacacg atgctttaac
     3421 tgcaaaaatt cagctcacca gttttgaggc aaaatttttg agtgacatgc aaagtaagca
     3481 tgatctcaat ggttcgttct catggctcac gcaaaaacaa cgaaccacac tagagaacat
     3541 actggctaaa tacggaagga tctgaggttc ttatggctct tgtatctatc agtgaagcat
     3601 caagactaac aaacaaaagt agaacaactg ttcaccgtta gatatcaaag ggaaaactgt
     3661 ccatatgcac agatgaaaac ggtgtaaaaa agatagatac atcagagctt ttacgagttt
     3721 ttggtgcatt taaagctgtt caccatgaac agatcgacaa tgtaactact agagctaaat
     3781 acattcaaat atctatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3841 aaaaaggaag aatatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3901 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgccgaaga
     3961 tcagttgggt gcacgtgtgg gttacatcga actggacctc aacagcggta agattcttga
     4021 gagttttcgc cccgaagaac gtttcccaat gatgagcact tttaaagttc tgctctgtgg
     4081 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     4141 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg acggcatgac
     4201 agtacgcgaa ttatgcagcg ctgccataac catgagtgat aacacggcgg ccaacttact
     4261 tctgacaacg atcggaggac cgaaggagct taccgctttt ttgcacaaca tgggtgatca
     4321 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4381 tgacaccacg atgcctgtag ctatggcaac aacgttgcgc aaactcttaa ctggcgaact
     4441 tcttactctc gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4501 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4561 tgagcgtggg tcccgcggta ttattgcagc cctggggcca gatggtaagc cctcccgtat
     4621 cgtagttatc tacacgacgg ggagccaggc aactatggac gaacgtaatc gccagatcgc
     4681 tgagataggt gcctccctga ttaagcattg gtaataatac tagctccggc aaaaaaacgg
     4741 gcaaggtgtc accaccctgc cctttttctt taaaaccgaa aagattactt cgcgtttgcc
     4801 acctgacgtc taagaaaagg aatattcagc aatttgcccg tgccgaagaa aggcccaccc
     4861 gtgaaggtga gccagtgagt tgattgctac gtaattagtt agttagccct tagtgactct
     4921 gccacctgac gtctaagaaa aggaatattc agcaatttgc ccgtgccgaa gaaaggccca
     4981 cccgtgaagg tgagccagtg agttgattgc tacgtaatta gttagttagc ccttagtgac
     5041 tc
//
