LOCUS       sequence-292316-        7431 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tunable NOR gate.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7431)
  AUTHORS   Bartoli V, Meaker GA, di Bernardo M, Gorochowski TE
  TITLE     Tunable genetic devices through simultaneous control of 
            transcription and translation.
  JOURNAL   Nat Commun. 2020 Apr 29;11(1):2095. doi: 10.1038/s41467-020-15653-7.
  PUBMED    32350250
REFERENCE   2  (bases 1 to 7431)
  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: "2020-04-29"; volume: "11(1):2095. doi"; pages: " 
            10.1038/s41467-020-15653-7"
COMMENT     Sequence Label: pVB006
FEATURES             Location/Qualifiers
     source          1..7431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(155..160,161..176,177..183)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    189..208
                     /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."
     promoter        347..631
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     554..573
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     protein_bind    672..690
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    697..715
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        728..777
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     misc_RNA        1678..1727
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             join(1777..1779,1780..1782,1783..2496)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDVTYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1822..1843)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2083..2102)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2430..2451
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        2606..2640
                     /label=BBa_J23105
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.24"
     RBS             2649..2660
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3516..3527
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             3645..4727
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(3664..3683)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4746..5369
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(4819..4838)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(5552..5571)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      complement(5936..6481)
                     /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(6016..6035)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6570..7385)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(6570..7385)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     7293..7312
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7377..7396
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 catgatgata tatttttatc ttgtgcaatg tacatcagag attttgagac acaaccaatt
       61 attgaaggcc tccctaacgg ggggcctttt tttgtttctg gtctcccgct tacttttcat
      121 actcccgccg cggccgcaac gatcgttggc tgtgttgaca attaatcatc ggctcgtata
      181 atgtgtggaa ttgtgagcgc tcacaattgg gtcatgactg ggacacgcca gtcatgagaa
      241 tacagacata aagcaagcca gagattaacg aagccaatta ttgaacaccc ttcggggtgt
      301 ttttttgttt ctggtctacc acttttcata ctcccgccat tcagagaaga aaccaattgt
      361 ccatattgca tcagacattg ccgtcactgc gtcttttact ggctcttctc gctaaccaaa
      421 ccggtaaccc cgcttattaa aagcattctg taacaaagcg ggaccaaagc catgacaaaa
      481 acgcgtaaca aaagtgtcta taatcacggc agaaaagtcc acattgatta tttgcacggc
      541 gtcacacttt gctatgccat agcattttta tccataagat tagcggatcc tacctgacgc
      601 tttttatcgc aactctctac tgtttctcca tacccgtttt tttgggctag ctactccacc
      661 gttggctttt ttccctatca gtgatagaga ttgacatccc tatcagtgat agagataatg
      721 agcacagctg tcaccggatg tgctttccgg tctgatgagt ccgtgaggac gaaacagcct
      781 ctacaaataa ttttgtttaa gggcgttaat ctctggcttg ctttatgtct gtaaacagag
      841 gagatacaga atgaaagcaa gcaacctggc ggcagcgcaa aagatgcgta aagcacgtac
      901 cccgagccgt agcagcattg gtagcctgcg tagtccgcat acccataaag caattctgac
      961 cagcaccatt gaaatcctga aagaatgtgg ttatagcggt ctgagcattg aaagcgttgc
     1021 acgtcgtgcc ggtgcaagca aaccgaccat ttatcgttgg tggaccaata aagcagcact
     1081 gattgccgaa gtgtatgaaa atgaaagcga acaggtgcgt aaatttccgg atctgggtag
     1141 ctttaaagcc gatctggatt ttctgctgcg taatctgtgg aaagtttggc gtgaaaccat
     1201 ttgtggtgaa gcatttcgtt gtgttattgc agaagcacag ctggaccctg caaccctgac
     1261 ccagctgaaa gatcagttta tggaacgtcg tcgtgagatg ccgaaaaaac tggttgaaaa
     1321 tgccattagc aatggtgaac tgccgaaaga taccaatcgt gaactgctgc tggatatgat
     1381 ttttggtttt tgttggtatc gcctgctgac cgaacagctg accgttgaac aggatattga
     1441 agaatttacc ttcctgctga ttaatggtgt ttgtccgggt acacagcgtt aaggaaacac
     1501 agaaaaaagc ccgcacctga cagtgcgggc ttcgatcaac tcgcgccgct ggtcttacgg
     1561 tctcatgaca gtgcgggctt tttttttcga ccaaaggtgt caacgttcga cgtacggtgg
     1621 aatctgattc gttaccaatt gacatgatac gaaacgtacc gtatcgttaa ggtcaagctg
     1681 tcaccggatg tgctttccgg tctgatgagt ccgtgaggac gaaacagcct ctacaaataa
     1741 ttttgtttaa tactagagaa agaggggaaa tactagatgg tgagcaaggg cgaggagctg
     1801 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     1861 agcgtgtccg gcgagggcga gggcgatgtc acctacggca agctgaccct gaagttcatc
     1921 tgcaccacag gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cggctacggc
     1981 ctgcaatgct tcgcccgcta ccccgaccac atgaagctgc acgacttctt caagtccgcc
     2041 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     2101 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     2161 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     2221 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     2281 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacacccca
     2341 atcggcgacg gccccgtgct gctgcccgac aaccactacc ttagctacca gtccgccctg
     2401 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     2461 gggatcactc tcggcatgga cgagctgtac aagtaagaat tccatggacg agctgtacaa
     2521 gtaactcggt accaaattcc agaaaagagg cctcccgaaa ggggggcctt ttttcgtttt
     2581 ggtccaatgg tcaccatata tcaagtttac ggctagctca gtcctaggta ctatgctagc
     2641 tactagagaa agaggagaaa tactagatgg ctgaagcgca aaatgatccc ctgctgccgg
     2701 gatactcgtt taatgcccat ctggtggcgg gtttaacgcc gattgaggcc aacggttatc
     2761 tcgatttttt tatcgaccga ccgctgggaa tgaaaggtta tattctcaat ctcaccattc
     2821 gcggtcaggg ggtggtgaaa aatcagggac gagaatttgt ttgccgaccg ggtgatattt
     2881 tgctgttccc gccaggagag attcatcact acggtcgtca tccggaggct cgcgaatggt
     2941 atcaccagtg ggtttacttt cgtccgcgcg cctactggca tgaatggctt aactggccgt
     3001 caatatttgc caatacgggg ttctttcgcc cggatgaagc gcaccagccg catttcagcg
     3061 acctgtttgg gcaaatcatt aacgccgggc aaggggaagg gcgctattcg gagctgctgg
     3121 cgataaatct gcttgagcaa ttgttactgc ggcgcatgga agcgattaac gagtcgctcc
     3181 atccaccgat ggataatcgg gtacgcgagg cttgtcagta catcagcgat cacctggcag
     3241 acagcaattt tgatatcgcc agcgtcgcac agcatgtttg cttgtcgccg tcgcgtctgt
     3301 cacatctttt ccgccagcag ttagggatta gcgtcttaag ctggcgcgag gaccaacgta
     3361 tcagccaggc gaagctgctt ttgagcacca cccggatgcc tatcgccacc gtcggtcgca
     3421 atgttggttt tgacgatcaa ctctatttct cgcgggtatt taaaaaatgc accggggcca
     3481 gcccgagcga gttccgtgcc ggttaataac caattattga aggccgctaa cgcagccttt
     3541 ttttgtttct ggtctcccaa tggcggcgcg ccatcgaatg gcgcaaaacc tttcgcggta
     3601 tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatatgaaa ccagtaacgt
     3661 tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc
     3721 aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga
     3781 attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg
     3841 ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc
     3901 gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag
     3961 cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact
     4021 atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt
     4081 tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc catgaggacg
     4141 gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag
     4201 cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca
     4261 ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt
     4321 ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca
     4381 acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg
     4441 cggatatctc ggtagtggga tacgacgata ccgaagatag ctcatgttat atcccgccgt
     4501 taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc
     4561 aactctctca gggccaggcg gtgaagggca atcagctgtt gccagtctca ctggtgaaaa
     4621 gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     4681 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgacaa tccaggagga
     4741 aaaaaatgtc cagattagat aaaagtaaag tgattaacag cgcattagag ctgcttaatg
     4801 aggtcggaat cgaaggttta acaacccgta aactcgccca gaagctaggt gtagagcagc
     4861 ctacattgta ttggcatgta aaaaataagc gggctttgct cgacgcctta gccattgaga
     4921 tgttagatag gcaccatact cacttttgcc ctttagaagg ggaaagctgg caagattttt
     4981 tacgtaataa cgctaaaagt tttagatgtg ctttactaag tcatcgcgat ggagcaaaag
     5041 tacatttagg tacacggcct acagaaaaac agtatgaaac tctcgaaaat caattagcct
     5101 ttttatgcca acaaggtttt tcactagaga atgcattata tgcactcagc gctgtggggc
     5161 attttacttt aggttgcgta ttggaagatc aagagcatca agtcgctaaa gaagaaaggg
     5221 aaacacctac tactgatagt atgccgccat tattacgaca agctatcgaa ttatttgatc
     5281 accaaggtgc agagccagcc ttcttattcg gccttgaatt gatcatatgc ggattagaaa
     5341 aacaacttaa atgtgaaagt gggtcctaat aattggtaac gaatcagaca attgacggct
     5401 cgagggagta gcatagggtt tgcagaatcc ctgcttcgtc catttgacag gcacattatg
     5461 catcgatgat aagctgtcaa acatgagcag atcctctacg ccggacgcat cgtggccggc
     5521 atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa
     5581 gatcgggctc gccacttcgg gctcatgagc aaatatttta tctgaggtgc ttcctcgctc
     5641 actgactcgc tgcacgaggc agacctcagc gctagcggag tgtatactgg cttactatgt
     5701 tggcactgat gagggtgtca gtgaagtgct tcatgtggca ggagaaaaaa ggctgcaccg
     5761 gtgcgtcagc agaatatgtg atacaggata tattccgctt cctcgctcac tgactcgcta
     5821 cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg aacggggcgg agatttcccg
     5881 gaagatgcca ggaagatact taacagggaa gtgagagggc cgcggcaaag ccgtttttcc
     5941 ataggctccg cccccctgac aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa
     6001 acccgacagg actataaaga taccaggcgt ttccccctgg cggctccctc gtgcgctctc
     6061 ctgttcctgc ctttcggttt accggtgtca ttccgctgtt atggccgcgt ttgtctcatt
     6121 ccacgcctga cactcagttc cgggtaggca gttcgctcca agctggactg tatgcacgaa
     6181 ccccccgttc agtccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6241 gaaagacatg caaaagcacc actggcagca gccactggta attgatttag aggagttagt
     6301 cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca agttttggtg actgcgctcc
     6361 tccaagccag ttacctcggt tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc
     6421 ctgcaaggcg gttttttcgt tttcagagca agagattacg cgcagaccaa aacgatctca
     6481 agaagatcat cttattaagg ggtctgacgc tcagtggaac gaaaaatcaa tctaaagtat
     6541 atatgagtaa acttggtctg acagttacct tagaaaaact catcgagcat caaatgaaac
     6601 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     6661 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     6721 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     6781 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagcttatgc
     6841 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     6901 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg
     6961 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
     7021 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg
     7081 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     7141 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     7201 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat
     7261 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     7321 tcagcatcca tgttggaatt taatcgcggc ctggagcaag acgtttcccg ttgaatatgg
     7381 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt t
//
