LOCUS       sequence-178104-        2912 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 2912)
  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 2912)
  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: pZA1pLtetO-GFP
FEATURES             Location/Qualifiers
     source          1..2912
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..19,20..25,26..42,43..48,49..74)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1..19
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    26..44
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             84..95
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(132..160)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     755..775
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      878..964
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(999..1016)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1133..1678)
                     /direction=LEFT
                     /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     complement(1213..1232)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1792..1886)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(1916..2707,2708..2776))
                     /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"
     primer_bind     2539..2558
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2777..2881)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tccctatcag tgatagagat tgacatccct atcagtgata gagatactga gcacatcagc
       61 aggacgcact gaccgaattc attaaagagg agaaaggtac catgagtaaa ggagaagaac
      121 ttttcactgg agttgtccca attcttgttg aattagatgg tgatgttaat gggcacaaat
      181 tttctgtcag tggagagggt gaaggtgatg caacatacgg aaaacttacc cttaaattta
      241 tttgcactac tggaaaacta cctgttccat ggccaacact tgtcactact ttcggttatg
      301 gtgttcaatg ctttgcgaga tacccagatc atatgaaaca gcatgacttt ttcaagagtg
      361 ccatgcccga aggttatgta caggaaagaa ctatattttt caaagatgac gggaactaca
      421 agacacgtgc tgaagtcaag tttgaaggtg atacccttgt taatagaatc gagttaaaag
      481 gtattgattt taaagaagat ggaaacattc ttggacacaa attggaatac aactataact
      541 cacacaatgt atacatcatg gcagacaaac aaaagaatgg aatcaaagtt aacttcaaaa
      601 ttagacacaa cattgaagat ggaagcgttc aactagcaga ccattatcaa caaaatactc
      661 caattggcga tggccctgtc cttttaccag acaaccatta cctgtccaca caatctgccc
      721 tttcgaaaga tcccaacgaa aagagagacc acatggtcct tcttgagttt gtaacagctg
      781 ctgggattac acatggcatg gatgatctct acaaataaaa gcttgatatc gaattcctgc
      841 agcccggggg atcccatggt acgcgtgcta gaggcatcaa ataaaacgaa aggctcagtc
      901 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
      961 aaatccgccg ccctagacct agggatatat tccgcttcct cgctcactga ctcgctacgc
     1021 tcggtcgttc gactgcggcg agcggaaatg gcttacgaac ggggcggaga tttcctggaa
     1081 gatgccagga agatacttaa cagggaagtg agagggccgc ggcaaagccg tttttccata
     1141 ggctccgccc ccctgacaag catcacgaaa tctgacgctc aaatcagtgg tggcgaaacc
     1201 cgacaggact ataaagatac caggcgtttc cccctggcgg ctccctcgtg cgctctcctg
     1261 ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg tctcattcca
     1321 cgcctgacac tcagttccgg gtaggcagtt cgctccaagc tggactgtat gcacgaaccc
     1381 cccgttcagt ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggaa
     1441 agacatgcaa aagcaccact ggcagcagcc actggtaatt gatttagagg agttagtctt
     1501 gaagtcatgc gccggttaag gctaaactga aaggacaagt tttggtgact gcgctcctcc
     1561 aagccagtta cctcggttca aagagttggt agctcagaga accttcgaaa aaccgccctg
     1621 caaggcggtt ttttcgtttt cagagcaaga gattacgcgc agaccaaaac gatctcaaga
     1681 agatcatctt attaatcaga taaaatattt ctagatttca gtgcaattta tctcttcaaa
     1741 tgtagcacct gaagtcagcc ccatacgata taagttgtta ctagtgcttg gattctcacc
     1801 aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga gttctgaggt
     1861 cattactgga tctatcaaca ggagtccaag cgagctcgta aacttggtct gacagttacc
     1921 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     1981 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     2041 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     2101 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     2161 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     2221 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     2281 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     2341 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     2401 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     2461 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     2521 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     2581 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     2641 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     2701 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     2761 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     2821 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     2881 gcacatttcc ccgaaaagtg ccacctgacg tc
//
