LOCUS       sequence-60652-e        5030 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 5030)
  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 5030)
  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: NOR gate-V2.0
FEATURES             Location/Qualifiers
     source          1..5030
                     /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      complement(129..156)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(172..243)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    302..351
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     protein_bind    466..513
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     misc_RNA        520..569
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             663..671
                     /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             678..1391
                     /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      1414..1485
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1501..1528
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1535..1555
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1591..1622
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     terminator      1747..1841
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2333..2555
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             2603..3553
                     /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(3842..3910,3911..4702)
                     /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(4832..4861)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     terminator      complement(4955..4984)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 gaattcgcgg ccgcttctag agaaaaaatt tatttgcttt cgcatctttt tgtacctata
       61 atgtgtggaa tgccaacaca attaacatct caatcaaggt aaatgctttt tgcttttttt
      121 gctataaacg cagaaaggcc cacccgaagg tgagccagtg tgactctagt agagagcgtt
      181 caccgacaaa caacagataa aacgaaaggc ccagtctttc gactgagcct ttcgttttat
      241 ttgatgcctg gaaaggagtt ttttagttac cttaattgaa ataaacgaaa taaaaactcg
      301 ctcggccggc ttgtcgacga cggcggtctc cgtcgtcagg atcatccggg ctttcttttg
      361 ggtatagcgt cgtggacagt cattcatctt tctgcccctc caaaagcaaa aacccgccga
      421 agcgggtttt tacgtaaatc aggtgaaact gaccgataag ccgggggttt gtaccgtaca
      481 ccactgagac cgcggtggtt gaccagacaa accacgaagc tgtcaccgga tgtgctttcc
      541 ggtctgatga gtccgtgagg acgaaacagc ctctacaaat aattttgttt aagggcccaa
      601 gttcacttaa aaaggagatc aacaatgaaa gcaattttcg tactgaaaca tcttaatcat
      661 gctaaggagg ttttctaatg cgtaaaggcg aagagctgtt cactggtgtc gtccctattc
      721 tggtggaact ggatggtgat gtcaacggtc ataagttttc cgtgcgtggc gagggtgaag
      781 gtgacgcaac taatggtaaa ctgacgctga agttcatctg tactactggt aaactgccgg
      841 taccttggcc gactctggta acgacgctga cttatggtgt tcagtgcttt gctcgttatc
      901 cggaccatat gaagcagcat gacttcttca agtccgccat gccggaaggc tatgtgcagg
      961 aacgcacgat ttcctttaag gatgacggca cgtacaaaac gcgtgcggaa gtgaaatttg
     1021 aaggcgatac tctggtaaac cgcattgagc tgaaaggcat tgactttaaa gaagacggca
     1081 atatcctggg ccataagctg gaatacaatt ttaacagcca caatgtttac atcaccgccg
     1141 ataaacaaaa aaatggcact aaagcgaatt ttaaaattcg ccacaacgtg gaggatggca
     1201 gcgtgcagct ggctgatcac taccagcaaa acactccaat cggtgatggt cctgttctgc
     1261 tgccagacaa tcactatctg agcacgcaaa gcgttctgtc taaagatccg aacgagaaac
     1321 gcgatcatat ggttctgctg gagttcgtaa ccgcagcggg catcacgcat ggtatggatg
     1381 aactgtacaa atgatgatac tagagccagg catcaaataa aacgaaaggc tcagtcgaaa
     1441 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gagtcacact
     1501 ggctcacctt cgggtgggcc tttctgcgtt tatatactag tagcggccgc tgcaggagtc
     1561 actaagggtt agttagttag attagcagaa agtcaaaagc ctccgaccgg aggcttttga
     1621 ctaaaacttc ccttggggtt atcattgggg ctcactcaaa ggcggtaatc agataaaaaa
     1681 aatccttagc tttcgctaag gatgatttct gctagagctg tcagaccaag tttacgagct
     1741 cgcttggact cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca
     1801 gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccaagcact agggacagta
     1861 agacgggtaa gcctgttgat gataccgctg ccttactggg tgcattagcc agtctgaatg
     1921 acctgtcacg ggataatccg aagtggtcag actggaaaat cagagggcag gaactgctga
     1981 acagcaaaaa gtcagatagc accacatagc agacccgcca taaaacgccc tgagaagccc
     2041 gtgacgggct tttcttgtat tatgggtagt ttccttgcat gaatccataa aaggcgcctg
     2101 tagtgccatt tacccccatt cactgccaga gccgtgagcg cagcgaactg aatgtcacga
     2161 aaaagacagc gactcaggtg cctgatggtc ggagacaaaa ggaatattca gcgatttgcc
     2221 cgagcttgcg agggtgctac ttaagccttt agggttttaa ggtctgtttt gtagaggagc
     2281 aaacagcgtt tgcgacatcc ttttgtaata ctgcggaact gactaaagta gtgagttata
     2341 cacagggctg ggatctattc tttttatctt tttttattct ttctttattc tataaattat
     2401 aaccacttga atataaacaa aaaaaacaca caaaggtcta gcggaattta cagagggtct
     2461 agcagaattt acaagttttc cagcaaaggt ctagcagaat ttacagatac ccacaactca
     2521 aaggaaaagg acatgtaatt atcattgact agcccatctc aattggtata gtgattaaaa
     2581 tcacctagac caattgagat gtatgtctga attagttgtt ttcaaagcaa atgaactagc
     2641 gattagtcgc tatgacttaa cggagcatga aaccaagcta attttatgct gtgtggcact
     2701 actcaacccc acgattgaaa accctacaag gaaagaacgg acggtatcgt tcacttataa
     2761 ccaatacgct cagatgatga acatcagtag ggaaaatgct tatggtgtat tagctaaagc
     2821 aaccagagag ctgatgacga gaactgtgga aatcaggaat cctttggtta aaggctttga
     2881 gattttccag tggacaaact atgccaagtt ctcaagcgaa aaattagaat tagtttttag
     2941 tgaagagata ttgccttatc ttttccagtt aaaaaaattc ataaaatata atctggaaca
     3001 tgttaagtct tttgaaaaca aatactctat gaggatttat gagtggttat taaaagaact
     3061 aacacaaaag aaaactcaca aggcaaatat agagattagc cttgatgaat ttaagttcat
     3121 gttaatgctt gaaaataact accatgagtt taaaaggctt aaccaatggg ttttgaaacc
     3181 aataagtaaa gatttaaaca cttacagcaa tatgaaattg gtggttgata agcgaggccg
     3241 cccgactgat acgttgattt tccaagttga actagataga caaatggatc tcgtaaccga
     3301 acttgagaac aaccagataa aaatgaatgg tgacaaaata ccaacaacca ttacatcaga
     3361 ttcctaccta cgtaacggac taagaaaaac actacacgat gctttaactg caaaaattca
     3421 gctcaccagt tttgaggcaa aatttttgag tgacatgcaa agtaagcatg atctcaatgg
     3481 ttcgttctca tggctcacgc aaaaacaacg aaccacacta gagaacatac tggctaaata
     3541 cggaaggatc tgaggttctt atggctcttg tatctatcag tgaagcatca agactaacaa
     3601 acaaaagtag aacaactgtt caccgttaga tatcaaaggg aaaactgtcc atatgcacag
     3661 atgaaaacgg tgtaaaaaag atagatacat cagagctttt acgagttttt ggtgcattta
     3721 aagctgttca ccatgaacag atcgacaatg taactactag agctaaatac attcaaatat
     3781 ctatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagaa
     3841 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     3901 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gccgaagatc agttgggtgc
     3961 acgtgtgggt tacatcgaac tggacctcaa cagcggtaag attcttgaga gttttcgccc
     4021 cgaagaacgt ttcccaatga tgagcacttt taaagttctg ctctgtggcg cggtattatc
     4081 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4141 ggttgagtac tcaccagtca cagaaaagca tcttacggac ggcatgacag tacgcgaatt
     4201 atgcagcgct gccataacca tgagtgataa cacggcggcc aacttacttc tgacaacgat
     4261 cggaggaccg aaggagctta ccgctttttt gcacaacatg ggtgatcatg taactcgcct
     4321 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     4381 gcctgtagct atggcaacaa cgttgcgcaa actcttaact ggcgaacttc ttactctcgc
     4441 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     4501 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     4561 ccgcggtatt attgcagccc tggggccaga tggtaagccc tcccgtatcg tagttatcta
     4621 cacgacgggg agccaggcaa ctatggacga acgtaatcgc cagatcgctg agataggtgc
     4681 ctccctgatt aagcattggt aataatacta gctccggcaa aaaaacgggc aaggtgtcac
     4741 caccctgccc tttttcttta aaaccgaaaa gattacttcg cgtttgccac ctgacgtcta
     4801 agaaaaggaa tattcagcaa tttgcccgtg ccgaagaaag gcccacccgt gaaggtgagc
     4861 cagtgagttg attgctacgt aattagttag ttagccctta gtgactctgc cacctgacgt
     4921 ctaagaaaag gaatattcag caatttgccc gtgccgaaga aaggcccacc cgtgaaggtg
     4981 agccagtgag ttgattgcta cgtaattagt tagttagccc ttagtgactc
//
