LOCUS       sequence-330509-        6742 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 6742)
  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 6742)
  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-CT82
FEATURES             Location/Qualifiers
     source          1..6742
                     /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"""
     promoter        23..57
                     /label=BBa_J23116
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.16"
     RBS             66..77
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             84..704
                     /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(157..176)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             705..737
                     /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      785..856
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      872..899
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             1052..1063
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1070..1783
                     /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             1070..1783
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGGTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1100..1128)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1723..1743
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1790..1822
                     /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      1845..1916
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1932..1959
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             2112..2123
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2130..3008
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     terminator      3030..3101
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3117..3144
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             3295..3306
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3892..3924
                     /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      3972..4043
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4059..4086
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    4093..4113
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(4253..5068)
                     /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(4253..5068)
                     /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     4976..4995
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5060..5079
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      complement(5663..6207)
                     /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(5742..5761)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6675..6693
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(6697..6740)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 gaattcgcgg ccgcttctag agttgacagc tagctcagtc ctagggacta tgctagctac
       61 tagagaaaga ggagaaatac tagatgtcca gattagataa aagtaaagtg attaacagcg
      121 cattagagct gcttaatgag gtcggaatcg aaggtttaac aacccgtaaa ctcgcccaga
      181 agctaggtgt agagcagcct acattgtatt ggcatgtaaa aaataagcgg gctttgctcg
      241 acgccttagc cattgagatg ttagataggc accatactca cttttgccct ttagaagggg
      301 aaagctggca agatttttta cgtaataacg ctaaaagttt tagatgtgct ttactaagtc
      361 atcgcgatgg agcaaaagta catttaggta cacggcctac agaaaaacag tatgaaactc
      421 tcgaaaatca attagccttt ttatgccaac aaggtttttc actagagaat gcattatatg
      481 cactcagcgc tgtggggcat tttactttag gttgcgtatt ggaagatcaa gagcatcaag
      541 tcgctaaaga agaaagggaa acacctacta ctgatagtat gccgccatta ttacgacaag
      601 ctatcgaatt atttgatcac caaggtgcag agccagcctt cttattcggc cttgaattga
      661 tcatatgcgg attagaaaaa caacttaaat gtgaaagtgg gtccgctgca aacgacgaaa
      721 actacgcttt agtagcttaa taacactgat agtgctagtg tagatcacta ctagagccag
      781 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
      841 gtcggtgaac gctctctact agagtcacac tggctcacct tcgggtgggc ctttctgcgt
      901 ttatatacta gagacattga ttatttgcac ggcgtcacac tttgctatgc catagcaaga
      961 tagtccataa gattagcgga tcctacctga cgctttttat cgcaactctc tactgtttct
     1021 ccataccgtt tttttgggct agctactaga gaaagaggag aaatactaga tgcgtaaagg
     1081 agaagaactt ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg
     1141 gcacaaattt tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct
     1201 taaatttatt tgcactactg gaaaactacc tgttccatgg ccaacacttg tcactacttt
     1261 cggttatggt gttcaatgct ttgcgagata cccagatcat atgaaacagc atgacttttt
     1321 caagagtgcc atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg
     1381 gaactacaag acacgtgctg aagtcaagtt tgaaggtggt acccttgtta atagaatcga
     1441 gttaaaaggt attgatttta aagaagatgg aaacattctt ggacacaaat tggaatacaa
     1501 ctataactca cacaatgtat acatcatggc agacaaacaa aagaatggaa tcaaagttaa
     1561 cttcaaaatt agacacaaca ttgaagatgg aagcgttcaa ctagcagacc attatcaaca
     1621 aaatactcca attggcgatg gccctgtcct tttaccagac aaccattacc tgtccacaca
     1681 atctgccctt tcgaaagatc ccaacgaaaa gagagaccac atggtccttc ttgagtttgt
     1741 aacagctgct gggattacac atggcatgga tgaactatac aaaaggcctg ctgcaaacga
     1801 cgaaaactac gctttagtag cttaataata ctagagccag gcatcaaata aaacgaaagg
     1861 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctctact
     1921 agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttatatacta gagacattga
     1981 ttatttgcac ggcgtcacac tttgctatgc catagcaaga tagtccataa gattagcgga
     2041 tcctacctga cgctttttat cgcaactctc tactgtttct ccataccgtt tttttgggct
     2101 agctactaga gaaagaggag aaatactaga tggctgaagc gcaaaatgat cccctgctgc
     2161 cgggatactc gtttaacgcc catctggtgg cgggtttaac gccgattgag gccaatggtt
     2221 atctcgattt ttttatcgac cgaccgctgg gaatgaaagg ttatattctc aatctcacca
     2281 ttcgcggtca gggggtggtg aaaaatcagg gacgagaatt tgtctgccga ccgggtgata
     2341 ttttgctgtt cccgccagga gagattcatc actacggtcg tcatccggag gctcgcgaat
     2401 ggtatcacca gtgggtttac tttcgtccgc gcgcctactg gcatgaatgg cttaactggc
     2461 cgtcaatatt tgccaatacg ggtttctttc gcccggatga agcgcaccag ccgcatttca
     2521 gcgacctgtt tgggcaaatc attaacgccg ggcaagggga agggcgctat tcggagctgc
     2581 tggcgataaa tctgcttgag caattgttac tgcggcgcat ggaagcgatt aacgagtcgc
     2641 tccatccacc gatggataat cgggtacgcg aggcttgtca gtacatcagc gatcacctgg
     2701 cagacagcaa ttttgatatc gccagcgtcg cacagcatgt ttgcctgtcg ccgtcgcgtc
     2761 tgtcacatct tttccgccag cagttaggga ttagcgtctt aagctggcgc gaggaccaac
     2821 gcatcagcca ggcgaagctg cttttgagca ctacccggat gcctatcgcc accgtcggtc
     2881 gcaatgttgg ttttgacgat caactctatt tctcgcgagt atttaaaaaa tgcaccgggg
     2941 ccagcccgag cgagttccgt gccggttgtg aagaaaaagt gaatgatgta gccgtcaagt
     3001 tgtcataata aactactaga gccaggcatc aaataaaacg aaaggctcag tcgaaagact
     3061 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct
     3121 caccttcggg tgggcctttc tgcgtttata tactagagac attgattatt tgcacggcgt
     3181 cacactttgc tatgccatag caagatagtc cataagatta gcggatccta cctgacgctt
     3241 tttatcgcaa ctctctactg tttctccatt ccctatcagt gatagatact agagaaagag
     3301 gagaaatact agatgactat aatgataaaa aaatcggatt ttttggcaat tccatcggag
     3361 gagtataaag gtattctaag tcttcgttat caagtgttta agcaaagact tgagtgggac
     3421 ttagttgtag aaaataacct tgaatcagat gagtatgata actcaaatgc agaatatatt
     3481 tatgcttgtg atgatactga aaatgtaagt ggatgctggc gtttattacc tacaacaggt
     3541 gattatatgc tgaaaagtgt ttttcctgaa ttgcttggtc aacagagtgc tcccaaagat
     3601 cctaatatag tcgaattaag tcgttttgct gtaggtaaaa atagctcaaa gataaataac
     3661 tctgctagtg aaattacaat gaaactattt gaagctatat ataaacacgc tgttagtcaa
     3721 ggtattacag aatatgtaac agtaacatca acagcaatag agcgattttt aaagcgtatt
     3781 aaagttcctt gtcatcgtat tggagacaaa gaaattcatg tattaggtga tactaaatcg
     3841 gttgtattgt ctatgcctat taatgaacag tttaaaaaag cagtcttaaa tgctgcaaac
     3901 gacgaaaact acgctttagt agcttaataa ctctgatagt gctagtgtag atctctacta
     3961 gagccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     4021 gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg ggtgggcctt
     4081 tctgcgttta tatactagta gcggccgctg caggcttcct cgctcactga ctcgctgcgc
     4141 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat ctcgagtccc
     4201 gtcaagtcag cgtaatgctc tgccagtgtt acaaccaatt aaccaattct gattagaaaa
     4261 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatcg ataccatatt
     4321 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     4381 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     4441 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
     4501 gtgagaatgg caaaagctta tgcatttctt tccagacttg ttcaacaggc cagccattac
     4561 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
     4621 cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
     4681 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
     4741 atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag
     4801 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
     4861 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
     4921 gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc
     4981 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctggagc
     5041 aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag
     5101 acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt
     5161 gagacacaac gtggctttgt tgaataaatc gaacttttgc tgagttgaag gatcagatca
     5221 cgcatcttcc cgacaacgca gaccgttccg tggcaaagca aaagttcaaa atcaccaact
     5281 ggtccaccta caacaaagct ctcatcaacc gtggctccct cactttctgg ctggatgatg
     5341 gggcgattca ggcctggtat gagtcagcaa caccttcttc acgaggcaga cctcagcgct
     5401 agcggagtgt atactggctt actatgttgg cactgatgag ggtgtcagtg aagtgcttca
     5461 tgtggcagga gaaaaaaggc tgcaccggtg cgtcagcaga atatgtgata caggatatat
     5521 tccgcttcct cgctcactga ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg
     5581 gcttacgaac ggggcggaga tttcctggaa gatgccagga agatacttaa cagggaagtg
     5641 agagggccgc ggcaaagccg tttttccata ggctccgccc cctgacaagc atcacgaaat
     5701 ctgacgctca aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5761 ccctggcggc tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc
     5821 gctgttatgg ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc
     5881 gctccaagct ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg
     5941 gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca
     6001 ctggtaattg atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa
     6061 aggacaagtt ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta
     6121 gctcagagaa ccttcaaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag
     6181 attacgcgca gaccaaaacg atctcaagaa gatcatctta ttaaggggtc tgacgctcag
     6241 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6301 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6361 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6421 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6481 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6541 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6601 gcctccatcc agtctattcc atggtgccac ctgacgtcta agaaaccatt attatcatga
     6661 cattaaccta taaaaatagg cgtatcacga ggcagaattt cagataaaaa aaatccttag
     6721 ctttcgctaa ggatgatttc tg
//
