LOCUS       sequence-208502-        7660 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CFP 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 7660)
  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 7660)
  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*500CFP
FEATURES             Location/Qualifiers
     source          1..7660
                     /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"
     CDS             complement(2006..2821)
                     /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(2006..2821)
                     /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     2729..2748
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      complement(3036..3063)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(3079..3150)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(3170..3883,3884..3886,3887..3889))
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3215..3236)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3564..3583
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3823..3844
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     RBS             3896..3907
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(6424..6441)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6558..7103)
                     /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(6638..6657)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7571..7589
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(7593..7636)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    7639..7660
                     /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 tcgctcgagt cccgtcaagt cagcgtaatg ctctgccagt
     1981 gttacaacca attaaccaat tctgattaga aaaactcatc gagcatcaaa tgaaactgca
     2041 atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag
     2101 gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc
     2161 cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa
     2221 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagc ttatgcattt
     2281 ccttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa
     2341 ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa
     2401 aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa
     2461 caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga
     2521 tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa
     2581 gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa
     2641 cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat
     2701 agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag
     2761 catccatgtt ggaatttaat cgcggcctgg agcaagacgt ttcccgttga atatggctca
     2821 taacaccact tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat
     2881 ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtggctt tgttgaataa
     2941 atcgaacttt tgctgagttg aaggatcaga tcacctacgt aattagttag ttagccctta
     3001 gtgactcgaa ttcgctgccg cttctagagt ataaacgcag aaaggcccac ccgaaggtga
     3061 gccagtgtga ctctagtaga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
     3121 gtctttcgac tgagcctttc gttttatttg atgcctggct ctagtattat cacttgtaca
     3181 gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat
     3241 cgcgcttctc gttggggtct ttgctcagct tggactgggt gctcaggtag tggttgtcgg
     3301 gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca
     3361 cgccgccgtc ctcgatgttg tggcggatct tgaagttggc cttgatgccg ttcttctgct
     3421 tgtcggcggt gatatagacg ttgtcgctaa agtagttgta ctccagcttg tgccccagga
     3481 tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc
     3541 cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc
     3601 gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt
     3661 cggggtagcg ggcgaagcac tgcacgcccc aggacagggt ggtcacgagg gtgggccagg
     3721 gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat
     3781 cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga
     3841 ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcaccatc tagtatttct
     3901 cctctttctc tagtattatt acgcgaacgc gaagtccgac tctaagatgt cacggaggtt
     3961 caagttacct ttagccggaa gtgctggcat tttgtccaat tgagactcgt gcaactggtc
     4021 agcgaactgg tcgtagaaat cagccagtac atcacaagac tcatatgtgt caaccatagt
     4081 ttcgcgcact gctttgaaca ggttcgcagc gtcagccgga atggtaccga aggagtcgtg
     4141 aatcagtgca aaagattcga ttccgtactt ctcgtgtgcc cacactacag tcttacgaag
     4201 gtggctaccg tcttggctgt gtacaaagtt aggagcgata ccagactcct gtttgtgtgc
     4261 atcaatctcg ctatctttgt tggtgttaat ggtaggctgt aagcggaact gaccgaggaa
     4321 catcaggttc aagcgcgtct gaataggctt cttgtattcc tgccacacag ggaaaccatc
     4381 aggagttacc caatgcacag cgcaacgctt gcgaagaatc tctccagtct tcttatcttt
     4441 gacctcagca gccagcagct tagcagcaga cttaagccag ttcattgctt caaccgcagc
     4501 taccaccgtc acgctcacag attcccaaat cagcttagcc atgtatccag cagcctgatt
     4561 cggctgagtg aacatcagac ccttgccgga atcaatagct ggctgaatgg tatcttccag
     4621 cacttgttga cggaagccga actctttgga cccgtaagcc agcgtcatga ctgaacgctt
     4681 agtcacactg cgagtaacac cgtaagccag ccattgacca gccagtgcct tagtgcccag
     4741 cttgactttc tcagagattt caccagtgtt ctcatcggtc acggtaacta cttcgttatc
     4801 ggtcccattg attgcgtctg cttgtagaat ctcgttgact ttcttagcaa caatcccgta
     4861 gatgtcctga acggtttcac taggaagcaa gttaaccgcg cgaccaccta cctcatctcg
     4921 gagcatcgcg gagaagtgct ggatgccaga gcaagacccg tcaaacgcca gcggaaggga
     4981 gcagttatag ctcaggccgt ggtgctgtac cccagcgtac tcaaagcaga acgcaaggaa
     5041 gcagaacgga gaatcttgct cagcccacca agtgttctcc agtggagact tagcgcaagc
     5101 catgatgttc tcgtggtttt cctcaatgaa cttgatgcgc tcagggaacg gaaccttatc
     5161 gacacccgca cagtttgcac cgtggatttt cagccagtag taaccttcct taccgattgg
     5221 tttacctttc gccagcgtaa gcagtccttt ggtcatatcg ttaccttgcg ggttgaacat
     5281 tgacacagcg taaacacgac cgcgccagtc catgttgtaa gggaaccaga tggccttatg
     5341 gttagcaaac ttattggctt gctcaagcat gaactcaagg ctgatacggc gagacttgcg
     5401 agccttgtcc ttgcggtaca cagcagcggc agcacgtttc cacgcggtga gagcctcagg
     5461 attcatgtcg atgtcttccg gtttcatcgg gagttcttca cgctcaatcg cagggatgtc
     5521 ctcgaccgga caatgcttcc acttggtgat tacgttggcg accgctagga ctttcttgtt
     5581 gattttccat gcggtgtttt gcgcaatgtt aatcgctttg tacacctcag gcatgtaaac
     5641 gtcttcgtag cgcatcagtg ctttcttact gtgagtacgc accagcgcca gaggacgacg
     5701 accgttagcc caatagccac caccagtaat gccagtccac ggcttaggag gaactacgca
     5761 aggttggaac atcggagaga tgccagccag cgcacctgca cgggttgcga tagcctcagc
     5821 gtattcaggt gcgagttcga tagtctcaga gtcttgacct actacgccag cattttggcg
     5881 gtgtaagcta accattccgg ttgactcaat gagcatctcg atgcagcgta ctcctacatg
     5941 aatagagtct tccttatgcc acgaagacca cgcctcgcca ccgagtagac ccttagagag
     6001 catgtcagcc tcgacaactt gcataaatgc tttcatctag tagtcctgtg tgactctagt
     6061 atttattcga aagtaacaaa ccattttctt gcgtaaactt gtacgatcct acaggtgcat
     6121 cttcccgaca acgcagaccg ttccgtggca aagcaaaagt tcaaaatcac caactggtcc
     6181 acctacaaca aagctctcat caaccgtggc tccctcactt tctggctgga tgatggggcg
     6241 attcgggcct ggtatgagtc agcaacacct tcttcacgag gcagacctca gcgctagcgg
     6301 agtgtatact ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg
     6361 caggagaaaa aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc
     6421 ttcctcgctc actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta
     6481 cgaacggggc ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg
     6541 gccgcggcaa agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatatga
     6601 cgctcaaatc agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6661 ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg
     6721 ttatggccgc gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc
     6781 caagctggac tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa
     6841 ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg
     6901 taattgattt agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga
     6961 caagttttgg tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc
     7021 agagaacctt cgaaaaaccg ccctgcaagg cggttttttc gttctcagag caagagatta
     7081 cgcgcagacc aaaacaatct caagaagatc atcttattaa ggggtctgac gctcagtgga
     7141 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7201 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7261 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7321 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7381 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7441 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7501 ccatccagtc tattccatgg tgccacctga cgtctaagaa accattatta tcatgacatt
     7561 aacctataaa aataggcgta tcacgaggca gaatttcaga taaaaaaaat ccttagcttt
     7621 cgctaaggat gatttctgga attcgcggcc gcttctagag
//
