LOCUS       sequence-330502-        5698 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacteria gene circuit on medium copy backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5698)
  AUTHORS   Zhang R, Goetz H, Melendez-Alvarez J, Li J, Ding T, Wang X, Tian XJ
  TITLE     Winner-takes-all resource competition redirects cascading cell fate 
            transitions.
  JOURNAL   Nat Commun. 2021 Feb 8;12(1):853. doi: 10.1038/s41467-021-21125-3.
  PUBMED    33558556
REFERENCE   2  (bases 1 to 5698)
  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: "2021-02-8"; volume: "12(1):853. doi"; pages: " 
            10.1038/s41467-021-21125-3"
COMMENT     Sequence Label: pSB3K3-CT66
FEATURES             Location/Qualifiers
     source          1..5698
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..22
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     RBS             86..97
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             104..982
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     terminator      1004..1075
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1091..1118
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1196..1207
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      1988..2059
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2075..2102
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             2180..2191
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2198..2872
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      2928..2999
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3015..3042
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    3049..3069
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(3209..4024)
                     /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(3209..4024)
                     /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     3932..3951
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4016..4035
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      complement(4619..5163)
                     /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(4698..4717)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5631..5649
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(5653..5696)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 gaattcgcgg ccgcttctag agacctgtag gatcgtacag ggttacgcaa gaaaatggtt
       61 tgttatagtc gaataaatac tagagaaaga ggagaaatac tagatggctg aagcgcaaaa
      121 tgatcccctg ctgccgggat actcgtttaa cgcccatctg gtggcgggtt taacgccgat
      181 tgaggccaat ggttatctcg atttttttat cgaccgaccg ctgggaatga aaggttatat
      241 tctcaatctc accattcgcg gtcagggggt ggtgaaaaat cagggacgag aatttgtctg
      301 ccgaccgggt gatattttgc tgttcccgcc aggagagatt catcactacg gtcgtcatcc
      361 ggaggctcgc gaatggtatc accagtgggt ttactttcgt ccgcgcgcct actggcatga
      421 atggcttaac tggccgtcaa tatttgccaa tacgggtttc tttcgcccgg atgaagcgca
      481 ccagccgcat ttcagcgacc tgtttgggca aatcattaac gccgggcaag gggaagggcg
      541 ctattcggag ctgctggcga taaatctgct tgagcaattg ttactgcggc gcatggaagc
      601 gattaacgag tcgctccatc caccgatgga taatcgggta cgcgaggctt gtcagtacat
      661 cagcgatcac ctggcagaca gcaattttga tatcgccagc gtcgcacagc atgtttgcct
      721 gtcgccgtcg cgtctgtcac atcttttccg ccagcagtta gggattagcg tcttaagctg
      781 gcgcgaggac caacgcatca gccaggcgaa gctgcttttg agcactaccc ggatgcctat
      841 cgccaccgtc ggtcgcaatg ttggttttga cgatcaactc tatttctcgc gagtatttaa
      901 aaaatgcacc ggggccagcc cgagcgagtt ccgtgccggt tgtgaagaaa aagtgaatga
      961 tgtagccgtc aagttgtcat aataaactac tagagccagg catcaaataa aacgaaaggc
     1021 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta
     1081 gagtcacact ggctcacctt cgggtgggcc tttctgcgtt tatatactag agacctgtag
     1141 gatcgtacag ggttacgcaa gaaaatggtt tgttatagtc gaataaatac tagagaaaga
     1201 ggagaaatac tagatgaaaa acataaatgc cgacgacaca tacagaataa ttaataaaat
     1261 taaagcttgt agaagcaata atgatattaa tcaatgctta tctgatatga ctaaaatggt
     1321 acattgtgaa tattatttac tcgcgatcat ttatcctcat tctatggtta aatctgatat
     1381 ttcaatccta gataattacc ctaaaaaatg gaggcaatat tatgatgacg ctaatttaat
     1441 aaaatatgat cctatagtag attattctaa ctccaatcat tcaccaatta attggaatat
     1501 atttgaaaac aatgctgtaa ataaaaaatc tccaaatgta attaaagaag cgaaaacagc
     1561 aggtcttatc actgggttta gtttccctat tcatacggct aacaatggct tcggaatact
     1621 tagttttgca cattcagaaa aagacaacta tatagatagt ttatttttac atgcgtgtat
     1681 gaacatacca ttaattgttc cttctctagt tgataattat cgaaaaataa atatagcaaa
     1741 taataaatca aacaacgatt taaccaaaag agaaaaagaa tgtttagcgt gggcatgcga
     1801 aggaaaaagc tcttgggata tttcaaaaat attaggttgc agtgagcgta ctgtcacttt
     1861 ccatttaacc aatgcgcaaa tgaaactcaa tacaacaaac cgctgccaaa gtatttctaa
     1921 agcaatttta acaggagcaa ttgattgccc atactttaaa aattaataaa ctactagagc
     1981 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     2041 tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg ggcctttctg
     2101 cgtttatata ctagagacct gtaggatcgt acagggttac gcaagaaaat ggtttgttat
     2161 agtcgaataa atactagaga aagaggagaa atactagatg gcttcctccg aagacgttat
     2221 caaagagttc atgcgtttca aagttcgtat ggaaggttcc gttaacggtc acgagttcga
     2281 aatcgaaggt gaaggtgaag gtcgtccgta cgaaggtacc cagaccgcta aactgaaagt
     2341 taccaaaggt ggtccgctgc cgttcgcttg ggacatcctg tccccgcagt tccagtacgg
     2401 ttccaaagct tacgttaaac acccggctga catcccggac tacctgaaac tgtccttccc
     2461 ggaaggtttc aaatgggaac gtgttatgaa cttcgaagac ggtggtgttg ttaccgttac
     2521 ccaggactcc tccctgcaag acggtgagtt catctacaaa gttaaactgc gtggtaccaa
     2581 cttcccgtcc gacggtccgg ttatgcagaa aaaaaccatg ggttgggaag cttccaccga
     2641 acgtatgtac ccggaagacg gtgctctgaa aggtgaaatc aaaatgcgtc tgaaactgaa
     2701 agacggtggt cactacgacg ctgaagttaa aaccacctac atggctaaaa aaccggttca
     2761 gctgccgggt gcttacaaaa ccgacatcaa actggacatc acctcccaca acgaagacta
     2821 caccatcgtt gaacagtacg aacgtgctga aggtcgtcac tccaccggtg ctgctgcaaa
     2881 cgacgaaaac tacgcagcag cagtttaata atactagagc caggcatcaa ataaaacgaa
     2941 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct
     3001 actagagtca cactggctca ccttcgggtg ggcctttctg cgtttatata ctagtagcgg
     3061 ccgctgcagg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3121 gtatcagctc actcaaaggc ggtaatctcg agtcccgtca agtcagcgta atgctctgcc
     3181 agtgttacaa ccaattaacc aattctgatt agaaaaactc atcgagcatc aaatgaaact
     3241 gcaatttatt catatcagga ttatcgatac catatttttg aaaaagccgt ttctgtaatg
     3301 aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga
     3361 ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat
     3421 caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa agcttatgca
     3481 tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat
     3541 caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt
     3601 taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat
     3661 caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg
     3721 ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg
     3781 gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg
     3841 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc
     3901 gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat
     3961 cagcatccat gttggaattt aatcgcggcc tggagcaaga cgtttcccgt tgaatatggc
     4021 tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata
     4081 tatttttatc ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttgttgaa
     4141 taaatcgaac ttttgctgag ttgaaggatc agatcacgca tcttcccgac aacgcagacc
     4201 gttccgtggc aaagcaaaag ttcaaaatca ccaactggtc cacctacaac aaagctctca
     4261 tcaaccgtgg ctccctcact ttctggctgg atgatggggc gattcaggcc tggtatgagt
     4321 cagcaacacc ttcttcacga ggcagacctc agcgctagcg gagtgtatac tggcttacta
     4381 tgttggcact gatgagggtg tcagtgaagt gcttcatgtg gcaggagaaa aaaggctgca
     4441 ccggtgcgtc agcagaatat gtgatacagg atatattccg cttcctcgct cactgactcg
     4501 ctacgctcgg tcgttcgact gcggcgagcg gaaatggctt acgaacgggg cggagatttc
     4561 ctggaagatg ccaggaagat acttaacagg gaagtgagag ggccgcggca aagccgtttt
     4621 tccataggct ccgccccctg acaagcatca cgaaatctga cgctcaaatc agtggtggcg
     4681 aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc tcgtgcgctc
     4741 tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc gtttgtctca
     4801 ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac tgtatgcacg
     4861 aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4921 cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt agaggagtta
     4981 gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg tgactgcgct
     5041 cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt caaaaaaccg
     5101 ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc aaaacgatct
     5161 caagaagatc atcttattaa ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5221 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5281 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5341 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5401 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5461 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5521 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattccatgg
     5581 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     5641 tcacgaggca gaatttcaga taaaaaaaat ccttagcttt cgctaaggat gatttctg
//
