LOCUS       sequence-97361-e        4269 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  used with pAND_YFP  to generate an AND gate without memory .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4269)
  AUTHORS   Yang L, Nielsen AA, Fernandez-Rodriguez J, McClune CJ, Laub MT, Lu 
            TK, Voigt CA
  TITLE     Permanent genetic memory with >1-byte capacity.
  JOURNAL   Nat Methods. 2014 Dec;11(12):1261-6. doi: 10.1038/nmeth.3147. Epub 
            2014 Oct 26.
  PUBMED    25344638
REFERENCE   2  (bases 1 to 4269)
  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: 
            "10.1038/nmeth.3147"; journalName: "Nat Methods"; date: "2014-12"; 
            volume: "11"; issue: "12"; pages: "1261-6"
COMMENT     Sequence Label: pSpec
FEATURES             Location/Qualifiers
     source          1..4269
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..792
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     protein_bind    964..982
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    989..1007
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             1045..1722
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1742..1813
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1829..1856
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      1993..2087
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2579..2801
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             2849..3799
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
        1 atgagggaag cggtgatcgc cgaagtatcg actcaactat cagaggtagt tggcgtcatc
       61 gagcgccatc tcgaaccgac gttgctggcc gtacatttgt acggctccgc agtggatggc
      121 ggcctgaagc cacacagtga tattgatttg ctggttacgg tgaccgtaag gcttgatgaa
      181 acaacgcggc gagctttgat caacgacctt ttggaaactt cggcttcccc tggagagagc
      241 gagattctcc gcgctgtaga agtcaccatt gttgtgcacg acgacatcat tccgtggcgt
      301 tatccagcta agcgcgaact gcaatttgga gaatggcagc gcaatgacat tcttgcaggt
      361 atcttcgagc cagccacgat cgacattgat ctggctatct tgctgacaaa agcaagagaa
      421 catagcgttg ccttggtagg tccagcggcg gaggaactct ttgatccggt tcctgaacag
      481 gatctatttg aggcgctaaa tgaaacctta acgctatgga actcgccgcc cgactgggct
      541 ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt ggtacagcgc agtaaccggc
      601 aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat
      661 cagcccgtca tacttgaagc tagacaggct tatcttggac aagaagaaga tcgcttggcc
      721 tcgcgcgcag atcagttgga agaatttgtc cactacgtga aaggcgagat caccaaggta
      781 gtcggcaaat aatactagct ccggcaaaaa aacgggcaag gtgtcaccac cctgcccttt
      841 ttctttaaaa ccgaaaagat tacttcgcgt ttgccacctg acgtctaaga attttcgaaa
      901 aaaggcctcc caaatcgggg ggcctttttt attgataaca aaatactcca ccgttggctt
      961 ttttccctat cagtgataga gattgacatc cctatcagtg atagagataa tgagcactac
     1021 tagagtcaca caggaaagta ctagatggct tcctccgaag acgttatcaa agagttcatg
     1081 cgtttcaaag ttcgtatgga aggttccgtt aacggtcacg agttcgaaat cgaaggtgaa
     1141 ggtgaaggtc gtccgtacga aggtacccag accgctaaac tgaaagttac caaaggtggt
     1201 ccgctgccgt tcgcttggga catcctgtcc ccgcagttcc agtacggttc caaagcttac
     1261 gttaaacacc cggctgacat cccggactac ctgaaactgt ccttcccgga aggtttcaaa
     1321 tgggaacgtg ttatgaactt cgaagacggt ggtgttgtta ccgttaccca ggactcctcc
     1381 ctgcaagacg gtgagttcat ctacaaagtt aaactgcgtg gtaccaactt cccgtccgac
     1441 ggtccggtta tgcagaaaaa aaccatgggt tgggaagctt ccaccgaacg tatgtacccg
     1501 gaagacggtg ctctgaaagg tgaaatcaaa atgcgtctga aactgaaaga cggtggtcac
     1561 tacgacgctg aagttaaaac cacctacatg gctaaaaaac cggttcagct gccgggtgct
     1621 tacaaaaccg acatcaaact ggacatcacc tcccacaacg aagactacac catcgttgaa
     1681 cagtacgaac gtgctgaagg tcgtcactcc accggtgctt aataatacta gagccaggca
     1741 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     1801 ggtgaacgct ctctactaga gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta
     1861 tagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     1921 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaactgtcag accaagttta
     1981 cgagctcgct tggactcctg ttgatagatc cagtaatgac ctcagaactc catctggatt
     2041 tgttcagaac gctcggttgc cgccgggcgt tttttattgg tgagaatcca agcactaggg
     2101 acagtaagac gggtaagcct gttgatgata ccgctgcctt actgggtgca ttagccagtc
     2161 tgaatgacct gtcacgggat aatccgaagt ggtcagactg gaaaatcaga gggcaggaac
     2221 tgctgaacag caaaaagtca gatagcacca catagcagac ccgccataaa acgccctgag
     2281 aagcccgtga cgggcttttc ttgtattatg ggtagtttcc ttgcatgaat ccataaaagg
     2341 cgcctgtagt gccatttacc cccattcact gccagagccg tgagcgcagc gaactgaatg
     2401 tcacgaaaaa gacagcgact caggtgcctg atggtcggag acaaaaggaa tattcagcga
     2461 tttgcccgag cttgcgaggg tgctacttaa gcctttaggg ttttaaggtc tgttttgtag
     2521 aggagcaaac agcgtttgcg acatcctttt gtaatactgc ggaactgact aaagtagtga
     2581 gttatacaca gggctgggat ctattctttt tatctttttt tattctttct ttattctata
     2641 aattataacc acttgaatat aaacaaaaaa aacacacaaa ggtctagcgg aatttacaga
     2701 gggtctagca gaatttacaa gttttccagc aaaggtctag cagaatttac agatacccac
     2761 aactcaaagg aaaaggacat gtaattatca ttgactagcc catctcaatt ggtatagtga
     2821 ttaaaatcac ctagaccaat tgagatgtat gtctgaatta gttgttttca aagcaaatga
     2881 actagcgatt agtcgctatg acttaacgga gcatgaaacc aagctaattt tatgctgtgt
     2941 ggcactactc aaccccacga ttgaaaaccc tacaaggaaa gaacggacgg tatcgttcac
     3001 ttataaccaa tacgctcaga tgatgaacat cagtagggaa aatgcttatg gtgtattagc
     3061 taaagcaacc agagagctga tgacgagaac tgtggaaatc aggaatcctt tggttaaagg
     3121 ctttgagatt ttccagtgga caaactatgc caagttctca agcgaaaaat tagaattagt
     3181 ttttagtgaa gagatattgc cttatctttt ccagttaaaa aaattcataa aatataatct
     3241 ggaacatgtt aagtcttttg aaaacaaata ctctatgagg atttatgagt ggttattaaa
     3301 agaactaaca caaaagaaaa ctcacaaggc aaatatagag attagccttg atgaatttaa
     3361 gttcatgtta atgcttgaaa ataactacca tgagtttaaa aggcttaacc aatgggtttt
     3421 gaaaccaata agtaaagatt taaacactta cagcaatatg aaattggtgg ttgataagcg
     3481 aggccgcccg actgatacgt tgattttcca agttgaacta gatagacaaa tggatctcgt
     3541 aaccgaactt gagaacaacc agataaaaat gaatggtgac aaaataccaa caaccattac
     3601 atcagattcc tacctacgta acggactaag aaaaacacta cacgatgctt taactgcaaa
     3661 aattcagctc accagttttg aggcaaaatt tttgagtgac atgcaaagta agcatgatct
     3721 caatggttcg ttctcatggc tcacgcaaaa acaacgaacc acactagaga acatactggc
     3781 taaatacgga aggatctgag gttcttatgg ctcttgtatc tatcagtgaa gcatcaagac
     3841 taacaaacaa aagtagaaca actgttcacc gttagatatc aaagggaaaa ctgtccatat
     3901 gcacagatga aaacggtgta aaaaagatag atacatcaga gcttttacga gtttttggtg
     3961 catttaaagc tgttcaccat gaacagatcg acaatgtaac gattatcaaa aaggatcttc
     4021 acctagatcc ttttaaatta aaaatgaagc ttcgttgtgt ctcaaaatct ctgatgttac
     4081 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
     4141 aatacaaggg gtgtttacta gaggttgatc gggcacgtaa gaggttccaa ctttcaccat
     4201 aatgaaataa gatcactacc gggcgtattt tttgagttat cgagattttc aggagctaag
     4261 gaagctaaa
//
