LOCUS       sequence-292673-        4937 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  XOR gate with IPTG and aTc as inputs and mRFP1 as output.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4937)
  AUTHORS   Rosado A, Cordero T, Rodrigo G
  TITLE     Binary addition in a living cell based on riboregulation.
  JOURNAL   PLoS Genet. 2018 Jul 19;14(7):e1007548. doi: 
            10.1371/journal.pgen.1007548. eCollection 2018 Jul.
  PUBMED    30024870
REFERENCE   2  (bases 1 to 4937)
  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: "PLoS 
            Genet."; date: "2018-07-19"; volume: "14(7):e1007548. doi"; pages: "
            10.1371/journal.pgen.1007548. eCollection 2018 Jul"
COMMENT     Sequence Label: pRHA40
FEATURES             Location/Qualifiers
     source          1..4937
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    820..836
                     /label=lac operator
                     /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)."
     protein_bind    875..893
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    900..918
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1136..1154
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1161..1179
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1257..1273
                     /label=lac operator
                     /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)."
     protein_bind    1280..1296
                     /label=lac operator
                     /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)."
     RBS             1393..1404
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1420..2094
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKMR
                     LKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHST
                     GA"
     terminator      2117..2148
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     primer_bind     complement(2272..2289)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2272..2288)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2281..2303)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2497..2516)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2616..2638
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2676..2694)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2762..2866
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2867..2935,2936..3727)
                     /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     complement(3085..3104)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3898..4486
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4387..4406
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4640..4657
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4774..4795
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4810..4815,4816..4833,4834..4840)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4848..4864
                     /label=lac operator
                     /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)."
     primer_bind     4853..4875
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4872..4888
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4872..4888
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gatccacaga aaagcccgcc tttcggtggg ctttgttaac cgaatgtttc ttcgggccac
       61 tggctcgcaa ttactttccc caccacgcta caggattcat tgcacggaat catgggatac
      121 tgcggattaa gcggttgcaa aaaaacttgt ccactgtcac ggataagttt cttaaaggta
      181 aactcatcgc ctccaagacg ggcgatgcaa aaatcccctg gttctacagc ctgttcgggg
      241 tcaaccaaaa tcagcatccc atccgggaat gagggtttac ttccagtggg tgccgtcatg
      301 gagttacctt ctacctccag ccaaaaagca gagtcagatg cctttttcgt ggtactcacc
      361 caacgctccg cgtcgccctt tgtaaacgta cgaagttcgg gactaaacat gcccgcttga
      421 acgtgggaga acaccgggta ttcatactcg ctacgtaagg acggttgcat agaaaccgcc
      481 tcatacatct cgtagatttc acgagcaatg gaaggactga actcttcaac acttactttc
      541 aggatttttg ctaataatgc tgcattgtat gcgtttaaag cattaatacc gttaaaaagc
      601 gcgcccactc cactctgccc catacccatt ttatccgcta cagactcttg ggataagcca
      661 agttcatttt tctttttctc gtagatggct ttcaaacgac gagcatcttc cagttgctcc
      721 tgcgtaagtg gctttttctt ggttgagggc cccataacca atatctcctc tttactggag
      781 gaagcaaaga agagatacta ggcaggggtg ctcagtatct tgttatccgc tcacaatgtc
      841 aatgttatcc gctcacattt atggtcagct cgagactcta tcattgatag agtttgacat
      901 ccctatcagt gatagagata ctgagcactg gaggagagag cgatcctagt tctcactcaa
      961 agaagggtag gactaggcag ggccgcaaaa aaccccgctt cggcggggtt ttttcgcctg
     1021 cagagatcta aaaaaaaagg ctccaaaagg agccgcctcg cataaatctg tcacagcatc
     1081 aacgtttccg ctgaactgtg acagactcac acaatcaacc ctcccgtgct cagtatctct
     1141 atcactgata gggatgtcaa tctctatcac tgatagggat ctagaaaaaa aaatccttag
     1201 ctttcgctaa ggatgcctcg cataatcaca caatcaaccc tcccgtgctc agtatcttgt
     1261 tatccgctca caatgtcaat tgttatccgc tcacaattag gccgcgaatt aacaccgtgc
     1321 gtgttgacta ttttacctct ggcggtgata atggttgccc tcgcataatt tcacttcttc
     1381 aatcctcccg ttaaagagga gaaattatga atgactagtg cttcctccga agacgttatc
     1441 aaagagttca tgcgtttcaa agttcgtatg gaaggttccg ttaacggtca cgagttcgaa
     1501 atcgaaggtg aaggtgaagg tcgtccgtac gaaggtaccc agaccgctaa actgaaagtt
     1561 accaaaggtg gtccgctgcc gttcgcttgg gacatcctgt ccccgcagtt ccagtacggt
     1621 tccaaagctt acgttaaaca cccggctgac atcccggact acctgaaact gtccttcccg
     1681 gaaggtttca aatgggaacg tgttatgaac ttcgaagacg gtggtgttgt taccgttacc
     1741 caggactcct ccctgcaaga cggtgagttc atctacaaag ttaaactgcg tggtaccaac
     1801 ttcccgtccg acggtccggt tatgcagaaa aaaaccatgg gttgggaagc ttccaccgaa
     1861 cgtatgtacc cggaagacgg tgctctgaaa ggtgaaatca aaatgcgtct gaaactgaaa
     1921 gacggtggtc actacgacgc tgaagttaaa accacctaca tggctaaaaa accggttcag
     1981 ctgccgggtg cttacaaaac cgacatcaaa ctggacatca cctcccacaa cgaagactac
     2041 accatcgttg aacagtacga acgtgctgaa ggtcgtcact ccaccggtgc ttaacaccga
     2101 aattcagtaa gcagaaagtc aaaagcctcc gaccggaggc ttttgactat tactcaacag
     2161 gtaaggcgcg aggttttcga attcatctgt cgcccgtctc aaacgcaacc ctccggcggc
     2221 ggcgatcggg tcagctcatc gatgaatcgc tatccatcgt catcgcgttg cactggccgt
     2281 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     2341 acatccccct ttcgccagct ggcgtaatag cgatgaggcc cgcaccgatc gcccttccca
     2401 acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct
     2461 gtgcggtatt tcacaccgca tttggtgcac tctcagtaca atctgctctg atgccgcata
     2521 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     2581 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     2641 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     2701 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     2761 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2821 acaataaccc tgataaatgc ttcaataata ttgaaaaagg tagagtatga gtattcaaca
     2881 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     2941 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     3001 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     3061 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     3121 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     3181 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     3241 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     3301 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     3361 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     3421 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     3481 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     3541 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     3601 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     3661 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     3721 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     3781 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     3841 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3901 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3961 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4021 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4081 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4141 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4201 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4261 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4321 aaaggcggac aggtatccgg taagcgacag ggtcggaaca ggagagcgca cgagggagct
     4381 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4441 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4501 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     4561 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     4621 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     4681 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     4741 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     4801 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     4861 acaatttcac acaggaaaca gctatgacca tgattacgcc acgagtcgca gcacagctcg
     4921 ctggtccaga actgatg
//
