LOCUS       sequence-73360-e        2619 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of AND gate in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2619)
  AUTHORS   Stanton BC, Nielsen AA, Tamsir A, Clancy K, Peterson T, Voigt CA
  TITLE     Genomic mining of prokaryotic repressors for orthogonal logic gates.
  JOURNAL   Nat Chem Biol. 2013 Dec 8. doi: 10.1038/nchembio.1411.
  PUBMED    24316737
REFERENCE   2  (bases 1 to 2619)
  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 Chem 
            Biol. 2013 Dec 8. doi: 10.1038/nchembio.1411."
COMMENT     Sequence Label: pAND
FEATURES             Location/Qualifiers
     source          1..2619
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    21..39
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    46..64
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      702..773
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      789..816
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        join(829..834,835..850,851..857)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    863..882
                     /label=lac operator (symmetric)
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     terminator      1542..1613
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1629..1656
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1738..1749
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(1756..1758,1759..1761,1762..2475)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1801..1822)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2062..2081)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2409..2430
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      2499..2570
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2586..2613
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 tactccaccg ttggcttttt tccctatcag tgatagagat tgacatccct atcagtgata
       61 gagataatga gcacgccatg ccattggctt ttgatagagg acaactacta gatgaacctg
      121 aaagataaaa ttctgggcgt tgccaaagaa ctgtttatca aaaatggcta taacgcaacc
      181 accaccggtg aaattgttaa actgagcgaa agcagcaaag gcaatctgta ttatcacttt
      241 aaaaccaaag agaacctgtt tctggaaatc ctgaacatcg aagaaagcaa atggcaagag
      301 cagtggaaaa aagaacaaat caaatgcaaa accaaccgcg agaaattcta tctgtataat
      361 gaactgagcc tgaccaccga atattactat ccgctgcaga atgccatcat cgagttttat
      421 accgagtact ataaaaccaa cagcatcaac gagaaaatga acaaactgga aaacaaatac
      481 atcgatgcct accacgtgat ctttaaagaa ggtaatctga acggcgaatg gtgcattaat
      541 gatgttaatg ccgtgagcaa aattgcagca aatgccgtta atggcattgt tacctttacc
      601 catgagcaga atatcaacga acgcattaaa ctgatgaaca aattcagcca gatctttctg
      661 aatggcctga gcaaataagg actgcgacgt tctccaggca tcaaataaaa cgaaaggctc
      721 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctactaga
      781 gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta tacgcttgtt gacaattaat
      841 catcggctcg tataatgtgt ggaattgtga gcgctcacaa ttctatggac tatgtttgaa
      901 agggagaaat actagatggc acgtaccccg agccgtagca gcattggtag cctgcgtagt
      961 ccgcataccc ataaagcaat tctgaccagc accattgaaa tcctgaaaga atgtggttat
     1021 agcggtctga gcattgaaag cgttgcacgt cgtgccggtg caagcaaacc gaccatttat
     1081 cgttggtgga ccaataaagc agcactgatt gccgaagtgt atgaaaatga aagcgaacag
     1141 gtgcgtaaat ttccggatct gggtagcttt aaagccgatc tggattttct gctgcgtaat
     1201 ctgtggaaag tttggcgtga aaccatttgt ggtgaagcat ttcgttgtgt tattgcagaa
     1261 gcacagctgg accctgcaac cctgacccag ctgaaagatc agtttatgga acgtcgtcgt
     1321 gagatgccga aaaaactggt tgaaaatgcc attagcaatg gtgaactgcc gaaagatacc
     1381 aatcgtgaac tgctgctgga tatgattttt ggtttttgtt ggtatcgcct gctgaccgaa
     1441 cagctgaccg ttgaacagga tattgaagaa tttaccttcc tgctgattaa tggtgtttgt
     1501 ccgggtacac agcgttaact agggcccata cccccaggca tcaaataaaa cgaaaggctc
     1561 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctactaga
     1621 gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta taggagagcg cgggtgagag
     1681 ggattcgtta ccaattgaca attgattgga cgttcaatat aatgctagct actagagaaa
     1741 gaggagaaat actagatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     1801 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     1861 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     1921 gtgccctggc ccaccctcgt gaccaccttc ggctacggcc tgcaatgctt cgcccgctac
     1981 cccgaccaca tgaagctgca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     2041 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     2101 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     2161 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     2221 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     2281 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     2341 ctgcccgaca accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag
     2401 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     2461 gagctgtaca agtaactagg gcccataccc ccaggcatca aataaaacga aaggctcagt
     2521 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc
     2581 acactggctc accttcgggt gggcctttct gcgtttata
//
