LOCUS       sequence-208503-        6118 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  YFP half-circuit controller plasmid [ca. 558.5 relative RBS strength
            at PLas81*].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6118)
  AUTHORS   Boehm CR, Grant PK, Haseloff J
  TITLE     Programmed hierarchical patterning of bacterial populations.
  JOURNAL   Nat Commun. 2018 Feb 22;9(1):776. doi: 10.1038/s41467-018-03069-3.
  PUBMED    29472537
REFERENCE   2  (bases 1 to 6118)
  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: "2018-02-22"; volume: "9(1):776. doi"; pages: " 
            10.1038/s41467-018-03069-3"
COMMENT     Sequence Label: pCRB DRT7VTPLux*500YFP
FEATURES             Location/Qualifiers
     source          1..6118
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..19
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    26..44
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      852..923
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      940..966
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    975..991
                     /label=lac operator
                     /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)."
     protein_bind    998..1014
                     /label=lac operator
                     /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)."
     terminator      1806..1852
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1894..1921
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             2575..2586
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(2593..2595,2596..2598,2599..3312)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2638..2659)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2899..2918)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3246..3267
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      3332..3403
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3419..3446
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3606..4421)
                     /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(3606..4421)
                     /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     4329..4348
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5016..5561)
                     /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(5096..5115)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6029..6047
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(6051..6094)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    6097..6118
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 tccctatcag tgatagagat tgacatccct atcagtgata gagatactga gcacatacta
       61 gagtcacaca ggactactag atgaaaaaca tcaacgcgga cgacacctac cgtatcatta
      121 acaaaatcaa agcgtgtcgt tctaacaacg atattaacca atgcctgtct gatatgacca
      181 aaatggttca ctgtgaatat tatctgctgg ccatcattta tccgcacagc atggtaaaat
      241 ccgatatttc tattctggat aactacccaa aaaaatggcg tcagtactat gatgacgcca
      301 acctgatcaa gtacgatccg atcgtagatt actctaactc taaccattcc ccgattaact
      361 ggaatatctt tgagaacaac gcggttaaca aaaagtcccc aaacgttatc aaagaagcaa
      421 aaaccagcgg tctgatcacc ggtttcagct tcccgattca taccgcgaac aacggcttcg
      481 gtatgctgtc ttttgcgcac tccgaaaaag acaactacat cgattctctg ttcctgcacg
      541 cgtgcatgaa cattcctctg atcgttccgt ccctggttga taactatcgc aagattaaca
      601 ttgccaataa caaaagcaac aacgacctga ccaaacgcga gaaagaatgc ctggcatggg
      661 cgtgcgaagg taagtcctct tgggacatct ccaaaattct gggttgctct gaacgcaccg
      721 taactttcca tctgaccaac gcgcagatga aactgaacac cacgaatcgt tgtcaatcta
      781 tcagcaaagc tatcctgacc ggtgcgatcg attgcccgta tttcaaaaac taataactaa
      841 tggccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
      901 gttgtttgtc ggtgaacgct ctctactaga gtcacacttg ctcaccttcg ggtgggcctt
      961 tctgcgttta taaattgtga gcggataaca attgacattg tgagcggata acaagatact
     1021 gagcacatac tagagggata ttaactaatt taacggcatc acgagtaatg gctctggtgg
     1081 atggttttct ggaactggaa cgttcttctg gtaaactgga atggagcgct atcctgcaaa
     1141 aaatggcctc tgacctgggt ttctctaaaa tcctgttcgg cctgctgccg aaagattccc
     1201 aagactacga aaacgcattc atcgtaggca attacccggc cgcgtggcgt gaacattatg
     1261 atcgtgccgg ttatgcacgt gtagatccaa ccgtttccca ttgcacccag tccgtgctgc
     1321 cgattttctg ggaaccgagc atctaccaga cccgtaaaca gcacgagttc ttcgaagagg
     1381 cgtccgcggc tggtctggtt tacggcctga cgatgccgct gcatggtgct cgtggcgaac
     1441 tgggcgctct gtctctgtcc gtagaagcag aaaaccgcgc ggaggctaac cgtttcatcg
     1501 aatctgtact gccgaccctg tggatgctga aagattatgc actgcaatcc ggcgcgggtc
     1561 tggcgttcga acacccagtt tctaaaccgg tagttctgac cagccgtgaa aaagaggtcc
     1621 tgcaatggtg cgctatcggc aaaacctcct gggagatttc tgtaatttgc aactgctctg
     1681 aagccaacgt caacttccac atgggtaaca ttcgtcgtaa attcggtgtt acttctcgcc
     1741 gcgttgcagc gattatggcc gttaacctgg gcctgattac tctgtaataa gcatgcacca
     1801 ggcatcaaat aaaacgaaag gctcagtcga aagacttggc ctttcgtttt atctgttgtt
     1861 tgtcggtgaa acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc
     1921 gtttatatac tagagtgagc tcgacctata ggatcgtata ggtttacgca agaaaatggt
     1981 ttgttacttt cgaataaata ctagagtcac acaggactac tagatgaaca cgattaacat
     2041 cgctaagaac gacttctctg acatcgaact ggctgctatc ccgttcaaca ctctggctga
     2101 ccattacggt gagcgtttag ctcgcgaaca gttggccctt gagcatgagt cttacgagat
     2161 gggtgaagca cgcttccgca agatgtttga gcgtcaactt aaagctggtg aggttgcgga
     2221 taacgctgcc gccaagcctc tcatcactac cctactccct aagatgattg cacgcatcaa
     2281 cgactggttt gaggaagtga aagctaagcg cggcaagcgc ccgacagcct tccagttcct
     2341 gcaagaaatc aagccggaag ccgtagcgta catcaccatt aagaccactc tggcttgcct
     2401 aaccagtgct gacaatacaa ccgttcaggc tgtagcaagc gcaatcggtc gggccattga
     2461 ggacgaggct cgcttcggtc gtatccgtga ccttgaagct aagcacttca agaaaaacgt
     2521 tgaggaacaa ctcaacaagc gcgtagggca cgtctacaag taataatact agagaaagag
     2581 gagaaatact agatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     2641 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     2701 gatgccacct acggcaagct gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg
     2761 ccctggccca ccctcgtgac caccctgggc tacggcctgc agtgcttcgc ccgctacccc
     2821 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     2881 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     2941 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     3001 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac
     3061 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc
     3121 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     3181 cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc
     3241 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     3301 ctgtacaagt aataatacta gagccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     3361 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctactaga gtcacactgg
     3421 ctcaccttcg ggtgggcctt tctgcgttta tatactagag atgcaggctt cctcgctcac
     3481 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3541 aatctcgagt cccgtcaagt cagcgtaatg ctctgccagt gttacaacca attaaccaat
     3601 tctgattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
     3661 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
     3721 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     3781 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     3841 acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ccttccagac ttgttcaaca
     3901 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
     3961 gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
     4021 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     4081 ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat
     4141 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
     4201 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     4261 agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc
     4321 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     4381 cgcggcctgg agcaagacgt ttcccgttga atatggctca taacaccact tgtattactg
     4441 tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa
     4501 catcagagat tttgagacac aacgtggctt tgttgaataa atcgaacttt tgctgagttg
     4561 aaggatcaga tcacgcatct tcccaacaac gcagaccgtt ccgtggcaaa gcaaaagttc
     4621 aaaatcacca actggtccac ctacaacaaa gctctcatca accgtggctc cctcactttc
     4681 tggctggatg atggggcgat tcgggcctgg tatgagtcag caacaccttc ttcacgaggc
     4741 agacctcagc gctagcggag tgtatactgg cttactatgt tggcactgat gagggtgtca
     4801 gtgaagtgct tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg
     4861 atacaggata tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg
     4921 gcgagcggaa atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact
     4981 taacagggaa gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac
     5041 aagcatcacg aaatatgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga
     5101 taccaggcgt ttccccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt
     5161 accggtgtca ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc
     5221 cgggtaggca gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg
     5281 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc
     5341 actggcagca gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt
     5401 aaggctaaac tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt
     5461 tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt
     5521 tctcagagca agagattacg cgcagaccaa aacgatctca agaagatcat cttattaagg
     5581 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5641 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5701 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5761 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5821 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5881 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5941 ctgcaacttt atccgcctcc atccagtcta ttccatggtg ccacctgacg tctaagaaac
     6001 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggcaga atttcagata
     6061 aaaaaaatcc ttagctttcg ctaaggatga tttctggaat tcgcggccgc ttctagag
//
