LOCUS       sequence-330505-        5853 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 5853)
  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 5853)
  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-CT67
FEATURES             Location/Qualifiers
     source          1..5853
                     /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             161..172
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             758..790
                     /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      838..909
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      925..952
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1105..1116
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1123..1836
                     /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             1123..1836
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGGTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1153..1181)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1776..1796
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1843..1875
                     /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      1898..1969
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1985..2012
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             2165..2176
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2183..3061
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     terminator      3083..3154
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3170..3197
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    3204..3224
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(3364..4179)
                     /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(3364..4179)
                     /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     4087..4106
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4171..4190
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      complement(4774..5318)
                     /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(4853..4872)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5786..5804
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(5808..5851)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 gaattcgcgg ccgcttctag agacattgat tatttgcacg gcgtcacact ttgctatgcc
       61 atagcaagat agtccataag attagcggat cctacctgac gctttttatc gcaactctct
      121 actgtttctc cataccgttt ttttgggcta gctactagag aaagaggaga aatactagat
      181 gactataatg ataaaaaaat cggatttttt ggcaattcca tcggaggagt ataaaggtat
      241 tctaagtctt cgttatcaag tgtttaagca aagacttgag tgggacttag ttgtagaaaa
      301 taaccttgaa tcagatgagt atgataactc aaatgcagaa tatatttatg cttgtgatga
      361 tactgaaaat gtaagtggat gctggcgttt attacctaca acaggtgatt atatgctgaa
      421 aagtgttttt cctgaattgc ttggtcaaca gagtgctccc aaagatccta atatagtcga
      481 attaagtcgt tttgctgtag gtaaaaatag ctcaaagata aataactctg ctagtgaaat
      541 tacaatgaaa ctatttgaag ctatatataa acacgctgtt agtcaaggta ttacagaata
      601 tgtaacagta acatcaacag caatagagcg atttttaaag cgtattaaag ttccttgtca
      661 tcgtattgga gacaaagaaa ttcatgtatt aggtgatact aaatcggttg tattgtctat
      721 gcctattaat gaacagttta aaaaagcagt cttaaatgct gcaaacgacg aaaactacgc
      781 tttagtagct taataactct gatagtgcta gtgtagatct ctactagagc caggcatcaa
      841 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
      901 aacgctctct actagagtca cactggctca ccttcgggtg ggcctttctg cgtttatata
      961 ctagagacat tgattatttg cacggcgtca cactttgcta tgccatagca agatagtcca
     1021 taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt tctccatacc
     1081 gtttttttgg gctagctact agagaaagag gagaaatact agatgcgtaa aggagaagaa
     1141 cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa
     1201 ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt
     1261 atttgcacta ctggaaaact acctgttcca tggccaacac ttgtcactac tttcggttat
     1321 ggtgttcaat gctttgcgag atacccagat catatgaaac agcatgactt tttcaagagt
     1381 gccatgcccg aaggttatgt acaggaaaga actatatttt tcaaagatga cgggaactac
     1441 aagacacgtg ctgaagtcaa gtttgaaggt ggtacccttg ttaatagaat cgagttaaaa
     1501 ggtattgatt ttaaagaaga tggaaacatt cttggacaca aattggaata caactataac
     1561 tcacacaatg tatacatcat ggcagacaaa caaaagaatg gaatcaaagt taacttcaaa
     1621 attagacaca acattgaaga tggaagcgtt caactagcag accattatca acaaaatact
     1681 ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc
     1741 ctttcgaaag atcccaacga aaagagagac cacatggtcc ttcttgagtt tgtaacagct
     1801 gctgggatta cacatggcat ggatgaacta tacaaaaggc ctgctgcaaa cgacgaaaac
     1861 tacgctttag tagcttaata atactagagc caggcatcaa ataaaacgaa aggctcagtc
     1921 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtca
     1981 cactggctca ccttcgggtg ggcctttctg cgtttatata ctagagacat tgattatttg
     2041 cacggcgtca cactttgcta tgccatagca agatagtcca taagattagc ggatcctacc
     2101 tgacgctttt tatcgcaact ctctactgtt tctccatacc gtttttttgg gctagctact
     2161 agagaaagag gagaaatact agatggctga agcgcaaaat gatcccctgc tgccgggata
     2221 ctcgtttaac gcccatctgg tggcgggttt aacgccgatt gaggccaatg gttatctcga
     2281 tttttttatc gaccgaccgc tgggaatgaa aggttatatt ctcaatctca ccattcgcgg
     2341 tcagggggtg gtgaaaaatc agggacgaga atttgtctgc cgaccgggtg atattttgct
     2401 gttcccgcca ggagagattc atcactacgg tcgtcatccg gaggctcgcg aatggtatca
     2461 ccagtgggtt tactttcgtc cgcgcgccta ctggcatgaa tggcttaact ggccgtcaat
     2521 atttgccaat acgggtttct ttcgcccgga tgaagcgcac cagccgcatt tcagcgacct
     2581 gtttgggcaa atcattaacg ccgggcaagg ggaagggcgc tattcggagc tgctggcgat
     2641 aaatctgctt gagcaattgt tactgcggcg catggaagcg attaacgagt cgctccatcc
     2701 accgatggat aatcgggtac gcgaggcttg tcagtacatc agcgatcacc tggcagacag
     2761 caattttgat atcgccagcg tcgcacagca tgtttgcctg tcgccgtcgc gtctgtcaca
     2821 tcttttccgc cagcagttag ggattagcgt cttaagctgg cgcgaggacc aacgcatcag
     2881 ccaggcgaag ctgcttttga gcactacccg gatgcctatc gccaccgtcg gtcgcaatgt
     2941 tggttttgac gatcaactct atttctcgcg agtatttaaa aaatgcaccg gggccagccc
     3001 gagcgagttc cgtgccggtt gtgaagaaaa agtgaatgat gtagccgtca agttgtcata
     3061 ataaactact agagccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
     3121 tcgttttatc tgttgtttgt cggtgaacgc tctctactag agtcacactg gctcaccttc
     3181 gggtgggcct ttctgcgttt atatactagt agcggccgct gcaggcttcc tcgctcactg
     3241 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3301 tctcgagtcc cgtcaagtca gcgtaatgct ctgccagtgt tacaaccaat taaccaattc
     3361 tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
     3421 gataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
     3481 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
     3541 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
     3601 gactgaatcc ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg
     3661 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
     3721 ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
     3781 cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg
     3841 atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc
     3901 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
     3961 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     4021 aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc
     4081 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
     4141 cggcctggag caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt
     4201 tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca
     4261 tcagagattt tgagacacaa cgtggctttg ttgaataaat cgaacttttg ctgagttgaa
     4321 ggatcagatc acgcatcttc ccgacaacgc agaccgttcc gtggcaaagc aaaagttcaa
     4381 aatcaccaac tggtccacct acaacaaagc tctcatcaac cgtggctccc tcactttctg
     4441 gctggatgat ggggcgattc aggcctggta tgagtcagca acaccttctt cacgaggcag
     4501 acctcagcgc tagcggagtg tatactggct tactatgttg gcactgatga gggtgtcagt
     4561 gaagtgcttc atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat
     4621 acaggatata ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt cgactgcggc
     4681 gagcggaaat ggcttacgaa cggggcggag atttcctgga agatgccagg aagatactta
     4741 acagggaagt gagagggccg cggcaaagcc gtttttccat aggctccgcc ccctgacaag
     4801 catcacgaaa tctgacgctc aaatcagtgg tggcgaaacc cgacaggact ataaagatac
     4861 caggcgtttc cccctggcgg ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc
     4921 ggtgtcattc cgctgttatg gccgcgtttg tctcattcca cgcctgacac tcagttccgg
     4981 gtaggcagtt cgctccaagc tggactgtat gcacgaaccc cccgttcagt ccgaccgctg
     5041 cgccttatcc ggtaactatc gtcttgagtc caacccggaa agacatgcaa aagcaccact
     5101 ggcagcagcc actggtaatt gatttagagg agttagtctt gaagtcatgc gccggttaag
     5161 gctaaactga aaggacaagt tttggtgact gcgctcctcc aagccagtta cctcggttca
     5221 aagagttggt agctcagaga accttcaaaa aaccgccctg caaggcggtt ttttcgtttt
     5281 cagagcaaga gattacgcgc agaccaaaac gatctcaaga agatcatctt attaaggggt
     5341 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5401 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5461 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5521 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5581 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5641 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5701 caactttatc cgcctccatc cagtctattc catggtgcca cctgacgtct aagaaaccat
     5761 tattatcatg acattaacct ataaaaatag gcgtatcacg aggcagaatt tcagataaaa
     5821 aaaatcctta gctttcgcta aggatgattt ctg
//
