LOCUS       sequence-178133-        3007 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic gene circuit.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3007)
  AUTHORS   Rubens JR, Selvaggio G, Lu TK
  TITLE     Synthetic mixed-signal computation in living cells.
  JOURNAL   Nat Commun. 2016 Jun 3;7:11658. doi: 10.1038/ncomms11658.
  PUBMED    27255669
REFERENCE   2  (bases 1 to 3007)
  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"; journalName: "Nat 
            Commun."; date: "2016-06-3"; volume: "7:11658. doi"; pages: " 
            10.1038/ncomms11658"
COMMENT     Sequence Label: Bxbi GFP MCP Reporter
FEATURES             Location/Qualifiers
     source          1..3007
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(31..59)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     654..674
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      777..863
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    883..931
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     primer_bind     complement(1104..1122)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1188..1290
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1291..1950
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1357..1376)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1971..2065
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2179..2724
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     2625..2644
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    2842..2899
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attP site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     misc_RNA        2908..2957
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             2990..3001
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atgagtaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt
       61 gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc aacatacgga
      121 aaacttaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg gccaacactt
      181 gtcactactt tcggttatgg tgttcaatgc tttgcgagat acccagatca tatgaaacag
      241 catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaagaac tatatttttc
      301 aaagatgacg ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga tacccttgtt
      361 aatagaatcg agttaaaagg tattgatttt aaagaagatg gaaacattct tggacacaaa
      421 ttggaataca actataactc acacaatgta tacatcatgg cagacaaaca aaagaatgga
      481 atcaaagtta acttcaaaat tagacacaac attgaagatg gaagcgttca actagcagac
      541 cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac
      601 ctgtccacac aatctgccct ttcgaaagat cccaacgaaa agagagacca catggtcctt
      661 cttgagtttg taacagctgc tgggattaca catggcatgg atgatctcta caaataaaag
      721 cttgatatcg aattcctgca gcccggggga tcccatggta cgcgtgctag aggcatcaaa
      781 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
      841 acgctctcct gagtaggaca aatccgccgc cctagaatcg atgcccggat gatcctgacg
      901 acggagaccg ccgtcgtcga caagccggcc gttaattaaa aagttaaaca aaattatttg
      961 tagagggaaa ccgttgtggt ctccctgaat atattatacg agccttatgc atgcccgtaa
     1021 agttatccag caaccactca tagacctagg gcagcagata gggacgacgt ggtgttagct
     1081 gtgctcgagg tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     1141 gataataatg gtttcttaga cgtcgatatc tggcgaaaat gagacgttga tcggcacgta
     1201 agaggttcca actttcacca taatgaaata agatcactac cgggcgtatt ttttgagtta
     1261 tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc
     1321 gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa
     1381 tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa
     1441 aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat
     1501 ccggaattcc gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct
     1561 tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac
     1621 gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac
     1681 ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg
     1741 gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt
     1801 ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag
     1861 gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag
     1921 tactgcgatg agtggcaggg cggggcgtaa tttgatatcg agctcgcttg gactcctgtt
     1981 gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg
     2041 ccgggcgttt tttattggtg agaatccaag cactagtaac aacttatatc gtatggggct
     2101 gacttcaggt gctacatttg aagagataaa ttgcactgaa atctagaaat attttatctg
     2161 attaataaga tgatcttctt gagatcgttt tggtctgcgc gtaatctctt gctctgaaaa
     2221 cgaaaaaacc gccttgcagg gcggtttttc gaaggttctc tgagctacca actctttgaa
     2281 ccgaggtaac tggcttggag gagcgcagtc accaaaactt gtcctttcag tttagcctta
     2341 accggcgcat gacttcaaga ctaactcctc taaatcaatt accagtggct gctgccagtg
     2401 gtgcttttgc atgtctttcc gggttggact caagacgata gttaccggat aaggcgcagc
     2461 ggtcggactg aacggggggt tcgtgcatac agtccagctt ggagcgaact gcctacccgg
     2521 aactgagtgt caggcgtgga atgagacaaa cgcggccata acagcggaat gacaccggta
     2581 aaccgaaagg caggaacagg agagcgcacg agggagccgc cagggggaaa cgcctggtat
     2641 ctttatagtc ctgtcgggtt tcgccaccac tgatttgagc gtcagatttc gtgatgcttg
     2701 tcaggggggc ggagcctatg gaaaaacggc tttgccgcgg ccctctcact tccctgttaa
     2761 gtatcttcct ggcatcttcc aggaaatctc cgccccgttc gtaagccatt tccgctcgcc
     2821 gcagtcgaac gaccggctag cgtcgtggtt tgtctggtca accaccgcgg tctcagtggt
     2881 gtacggtaca aaccccgacg tgcacagctg tcaccggatg tgctttccgg tctgatgagt
     2941 ccgtgaggac gaaacagcct ctacaaataa ttttgtttaa gaattcatta aagaggagaa
     3001 aggtacc
//
