LOCUS       sequence-330511-        6833 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 6833)
  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 6833)
  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-IC15
FEATURES             Location/Qualifiers
     source          1..6833
                     /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)"
     terminator      878..949
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      965..992
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1070..1081
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1088..1762
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1818..1889
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1905..1932
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             2085..2096
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2103..2816
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGGTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             2103..2816
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGGTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2133..2161)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2756..2776
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             2823..2855
                     /codon_start=1
                     /product="C-terminal peptide that mediates degradation in 
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
                     /label=ssrA tag (LVA)
                     /note="mutant LVA variant that confers accelerated 
                     degradation under some conditions (Andersen et al., 1998)"
                     /translation="AANDENYALVA"
     terminator      2878..2949
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2965..2992
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             3145..3156
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3163..4041
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     terminator      4063..4134
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4150..4177
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    4184..4204
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(4344..5159)
                     /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(4344..5159)
                     /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     5067..5086
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5151..5170
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      complement(5754..6298)
                     /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(5833..5852)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6766..6784
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(6788..6831)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 gaattcgcgg ccgcttctag agacctgtag gatcgtacag ggttacgcaa gaaaatggtt
       61 tgttatagtc gaataaatac tagagaaaga ggagaaatac tagatgaaaa acataaatgc
      121 cgacgacaca tacagaataa ttaataaaat taaagcttgt agaagcaata atgatattaa
      181 tcaatgctta tctgatatga ctaaaatggt acattgtgaa tattatttac tcgcgatcat
      241 ttatcctcat tctatggtta aatctgatat ttcaatccta gataattacc ctaaaaaatg
      301 gaggcaatat tatgatgacg ctaatttaat aaaatatgat cctatagtag attattctaa
      361 ctccaatcat tcaccaatta attggaatat atttgaaaac aatgctgtaa ataaaaaatc
      421 tccaaatgta attaaagaag cgaaaacagc aggtcttatc actgggttta gtttccctat
      481 tcatacggct aacaatggct tcggaatact tagttttgca cattcagaaa aagacaacta
      541 tatagatagt ttatttttac atgcgtgtat gaacatacca ttaattgttc cttctctagt
      601 tgataattat cgaaaaataa atatagcaaa taataaatca aacaacgatt taaccaaaag
      661 agaaaaagaa tgtttagcgt gggcatgcga aggaaaaagc tcttgggata tttcaaaaat
      721 attaggttgc agtgagcgta ctgtcacttt ccatttaacc aatgcgcaaa tgaaactcaa
      781 tacaacaaac cgctgccaaa gtatttctaa agcaatttta acaggagcaa ttgattgccc
      841 atactttaaa aattaataaa ctactagagc caggcatcaa ataaaacgaa aggctcagtc
      901 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca
      961 cactggctca ccttcgggtg ggcctttctg cgtttatata ctagagacct gtaggatcgt
     1021 acagggttac gcaagaaaat ggtttgttat agtcgaataa atactagaga aagaggagaa
     1081 atactagatg gcttcctccg aagacgttat caaagagttc atgcgtttca aagttcgtat
     1141 ggaaggttcc gttaacggtc acgagttcga aatcgaaggt gaaggtgaag gtcgtccgta
     1201 cgaaggtacc cagaccgcta aactgaaagt taccaaaggt ggtccgctgc cgttcgcttg
     1261 ggacatcctg tccccgcagt tccagtacgg ttccaaagct tacgttaaac acccggctga
     1321 catcccggac tacctgaaac tgtccttccc ggaaggtttc aaatgggaac gtgttatgaa
     1381 cttcgaagac ggtggtgttg ttaccgttac ccaggactcc tccctgcaag acggtgagtt
     1441 catctacaaa gttaaactgc gtggtaccaa cttcccgtcc gacggtccgg ttatgcagaa
     1501 aaaaaccatg ggttgggaag cttccaccga acgtatgtac ccggaagacg gtgctctgaa
     1561 aggtgaaatc aaaatgcgtc tgaaactgaa agacggtggt cactacgacg ctgaagttaa
     1621 aaccacctac atggctaaaa aaccggttca gctgccgggt gcttacaaaa ccgacatcaa
     1681 actggacatc acctcccaca acgaagacta caccatcgtt gaacagtacg aacgtgctga
     1741 aggtcgtcac tccaccggtg ctgctgcaaa cgacgaaaac tacgcagcag cagtttaata
     1801 atactagagc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     1861 ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg
     1921 ggcctttctg cgtttatata ctagagacat tgattatttg cacggcgtca cactttgcta
     1981 tgccatagca agatagtcca taagattagc ggatcctacc tgacgctttt tatcgcaact
     2041 ctctactgtt tctccatacc gtttttttgg gctagctact agagaaagag gagaaatact
     2101 agatgcgtaa aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg
     2161 gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg
     2221 gaaaacttac ccttaaattt atttgcacta ctggaaaact acctgttcca tggccaacac
     2281 ttgtcactac tttcggttat ggtgttcaat gctttgcgag atacccagat catatgaaac
     2341 agcatgactt tttcaagagt gccatgcccg aaggttatgt acaggaaaga actatatttt
     2401 tcaaagatga cgggaactac aagacacgtg ctgaagtcaa gtttgaaggt ggtacccttg
     2461 ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt cttggacaca
     2521 aattggaata caactataac tcacacaatg tatacatcat ggcagacaaa caaaagaatg
     2581 gaatcaaagt taacttcaaa attagacaca acattgaaga tggaagcgtt caactagcag
     2641 accattatca acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt
     2701 acctgtccac acaatctgcc ctttcgaaag atcccaacga aaagagagac cacatggtcc
     2761 ttcttgagtt tgtaacagct gctgggatta cacatggcat ggatgaacta tacaaaaggc
     2821 ctgctgcaaa cgacgaaaac tacgctttag tagcttaata atactagagc caggcatcaa
     2881 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     2941 aacgctctct actagagtca cactggctca ccttcgggtg ggcctttctg cgtttatata
     3001 ctagagacat tgattatttg cacggcgtca cactttgcta tgccatagca agatagtcca
     3061 taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt tctccatacc
     3121 gtttttttgg gctagctact agagaaagag gagaaatact agatggctga agcgcaaaat
     3181 gatcccctgc tgccgggata ctcgtttaac gcccatctgg tggcgggttt aacgccgatt
     3241 gaggccaatg gttatctcga tttttttatc gaccgaccgc tgggaatgaa aggttatatt
     3301 ctcaatctca ccattcgcgg tcagggggtg gtgaaaaatc agggacgaga atttgtctgc
     3361 cgaccgggtg atattttgct gttcccgcca ggagagattc atcactacgg tcgtcatccg
     3421 gaggctcgcg aatggtatca ccagtgggtt tactttcgtc cgcgcgccta ctggcatgaa
     3481 tggcttaact ggccgtcaat atttgccaat acgggtttct ttcgcccgga tgaagcgcac
     3541 cagccgcatt tcagcgacct gtttgggcaa atcattaacg ccgggcaagg ggaagggcgc
     3601 tattcggagc tgctggcgat aaatctgctt gagcaattgt tactgcggcg catggaagcg
     3661 attaacgagt cgctccatcc accgatggat aatcgggtac gcgaggcttg tcagtacatc
     3721 agcgatcacc tggcagacag caattttgat atcgccagcg tcgcacagca tgtttgcctg
     3781 tcgccgtcgc gtctgtcaca tcttttccgc cagcagttag ggattagcgt cttaagctgg
     3841 cgcgaggacc aacgcatcag ccaggcgaag ctgcttttga gcactacccg gatgcctatc
     3901 gccaccgtcg gtcgcaatgt tggttttgac gatcaactct atttctcgcg agtatttaaa
     3961 aaatgcaccg gggccagccc gagcgagttc cgtgccggtt gtgaagaaaa agtgaatgat
     4021 gtagccgtca agttgtcata ataaactact agagccaggc atcaaataaa acgaaaggct
     4081 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag
     4141 agtcacactg gctcaccttc gggtgggcct ttctgcgttt atatactagt agcggccgct
     4201 gcaggcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4261 agctcactca aaggcggtaa tctcgagtcc cgtcaagtca gcgtaatgct ctgccagtgt
     4321 tacaaccaat taaccaattc tgattagaaa aactcatcga gcatcaaatg aaactgcaat
     4381 ttattcatat caggattatc gataccatat ttttgaaaaa gccgtttctg taatgaagga
     4441 gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg
     4501 actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt
     4561 gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagctt atgcatttct
     4621 ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc
     4681 aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa
     4741 ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca
     4801 atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc
     4861 gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga
     4921 ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg
     4981 ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag
     5041 attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca
     5101 tccatgttgg aatttaatcg cggcctggag caagacgttt cccgttgaat atggctcata
     5161 acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga tgatatattt
     5221 ttatcttgtg caatgtaaca tcagagattt tgagacacaa cgtggctttg ttgaataaat
     5281 cgaacttttg ctgagttgaa ggatcagatc acgcatcttc ccgacaacgc agaccgttcc
     5341 gtggcaaagc aaaagttcaa aatcaccaac tggtccacct acaacaaagc tctcatcaac
     5401 cgtggctccc tcactttctg gctggatgat ggggcgattc aggcctggta tgagtcagca
     5461 acaccttctt cacgaggcag acctcagcgc tagcggagtg tatactggct tactatgttg
     5521 gcactgatga gggtgtcagt gaagtgcttc atgtggcagg agaaaaaagg ctgcaccggt
     5581 gcgtcagcag aatatgtgat acaggatata ttccgcttcc tcgctcactg actcgctacg
     5641 ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa cggggcggag atttcctgga
     5701 agatgccagg aagatactta acagggaagt gagagggccg cggcaaagcc gtttttccat
     5761 aggctccgcc ccctgacaag catcacgaaa tctgacgctc aaatcagtgg tggcgaaacc
     5821 cgacaggact ataaagatac caggcgtttc cccctggcgg ctccctcgtg cgctctcctg
     5881 ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg tctcattcca
     5941 cgcctgacac tcagttccgg gtaggcagtt cgctccaagc tggactgtat gcacgaaccc
     6001 cccgttcagt ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggaa
     6061 agacatgcaa aagcaccact ggcagcagcc actggtaatt gatttagagg agttagtctt
     6121 gaagtcatgc gccggttaag gctaaactga aaggacaagt tttggtgact gcgctcctcc
     6181 aagccagtta cctcggttca aagagttggt agctcagaga accttcaaaa aaccgccctg
     6241 caaggcggtt ttttcgtttt cagagcaaga gattacgcgc agaccaaaac gatctcaaga
     6301 agatcatctt attaaggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6361 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6421 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6481 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6541 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6601 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6661 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctattc catggtgcca
     6721 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     6781 aggcagaatt tcagataaaa aaaatcctta gctttcgcta aggatgattt ctg
//
