LOCUS       sequence-203739-        5256 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ratiometric reporter [PT7(-3G)-RBSB0034, low copy number, LacI 
            generator removed].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5256)
  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 5256)
  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: p4g3VT
FEATURES             Location/Qualifiers
     source          1..5256
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             19..969
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     RBS             1233..1244
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(1251..1253,1254..1256,1257..1970)
                     /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"
     terminator      1990..2061
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2077..2104
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             join(2198..2266,2267..3058)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     terminator      complement(3188..3217)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     misc_feature    3264..3285
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     terminator      complement(3292..3319)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(3335..3406)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(3426..4139,4140..4142,4143..4145))
                     /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"
     RBS             4152..4163
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(4172..4206)
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     misc_feature    4207..4227
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      4263..4294
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     terminator      4419..4513
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      5005..5227
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 ctagaccaat tgagatgtat gtctgaatta gttgttttca aagcaaatga actagcgatt
       61 agtcgctatg acttaacgga gcatgaaacc aagctaattt tatgctgtgt ggcactactc
      121 aaccccacga ttgaaaaccc tacaaggaaa gaacggacgg tatcgttcac ttataaccaa
      181 tacgctcaga tgatgaacat cagtagggaa aatgcttatg gtgtattagc taaagcaacc
      241 agagagctga tgacgagaac tgtggaaatc aggaatcctt tggttaaagg ctttgagatt
      301 ttccagtgga caaactatgc caagttctca agcgaaaaat tagaattagt ttttagtgaa
      361 gagatattgc cttatctttt ccagttaaaa aaattcataa aatataatct ggaacatgtt
      421 aagtcttttg aaaacaaata ctctatgagg atttatgagt ggttattaaa agaactaaca
      481 caaaagaaaa ctcacaaggc aaatatagag attagccttg atgaatttaa gttcatgtta
      541 atgcttgaaa ataactacca tgagtttaaa aggcttaacc aatgggtttt gaaaccaata
      601 agtaaagatt taaacactta cagcaatatg aaattggtgg ttgataagcg aggccgcccg
      661 actgatacgt tgattttcca agttgaacta gatagacaaa tggatctcgt aaccgaactt
      721 gagaacaacc agataaaaat gaatggtgac aaaataccaa caaccattac atcagattcc
      781 tacctacata acggactaag aaaaacacta cacgatgctt taactgcaaa aattcagctc
      841 accagttttg aggcaaaatt tttgagtgac atgcaaagta agtatgatct caatggttcg
      901 ttctcatggc tcacgcaaaa acaacgaacc acactagaga acatactggc taaatacgga
      961 aggatctgag gttcttatgg ctcttgtatc tatcagtgaa gcatcaagac taacaaacaa
     1021 aagtagaaca actgttcacc gttacatatc aaagggaaaa ctgtccatat gcacagatga
     1081 aaacggtgta aaaaagatag atacatcaga gcttttacga gtttttggtg cattcaaagc
     1141 tgttcaccat gaacagatcg acaatgtaac tactagagta atacgactca ctgtagggaa
     1201 tacaagctac ttgttctttt tgcatactag agaaagagga gaaatactag atggtgagca
     1261 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     1321 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     1381 ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     1441 ccctgggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
     1501 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     1561 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1621 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1681 acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
     1741 ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc
     1801 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
     1861 accagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1921 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa taatactaga
     1981 gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     2041 tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc
     2101 tgcgtttata tactagagct aaatacattc aaatatctat ccgctcatga gacaataacc
     2161 ctgataaatg cttcaataat attgaaaaag gaagaatatg agtattcaac atttccgtgt
     2221 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     2281 ggtgaaagta aaagatgccg aagatcagtt gggtgcacgt gtgggttaca tcgaactgga
     2341 cctcaacagc ggtaagattc ttgagagttt tcgccccgaa gaacgtttcc caatgatgag
     2401 cacttttaaa gttctgctct gtggcgcggt attatcccgt attgacgccg ggcaagagca
     2461 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     2521 aaagcatctt acggacggca tgacagtacg cgaattatgc agcgctgcca taaccatgag
     2581 tgataacacg gcggccaact tacttctgac aacgatcgga ggaccgaagg agcttaccgc
     2641 ttttttgcac aacatgggtg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     2701 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagctatgg caacaacgtt
     2761 gcgcaaactc ttaactggcg aacttcttac tctcgcttcc cggcaacaat taatagactg
     2821 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     2881 tattgctgat aaatctggag ccggtgagcg tgggtcccgc ggtattattg cagccctggg
     2941 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagcc aggcaactat
     3001 ggacgaacgt aatcgccaga tcgctgagat aggtgcctcc ctgattaagc attggtaata
     3061 atactagctc cggcaaaaaa acgggcaagg tgtcaccacc ctgccctttt tctttaaaac
     3121 cgaaaagatt acttcgcgtt tgccacctga cgtctaagaa aaggaatatt cagcaatttg
     3181 cccgtgccga agaaaggccc acccgtgaag gtgagccagt gagttgattg ctacgtaatt
     3241 agttagttag cccttagtga ctcgaattcg cggccgcttc tagagtataa acgcagaaag
     3301 gcccacccga aggtgagcca gtgtgactct agtagagagc gttcaccgac aaacaacaga
     3361 taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggctctag
     3421 tattattact tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc
     3481 agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagcttgga ctgggtgctc
     3541 aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag
     3601 tggtcggcga gctgcacgcc gccgtcctcg atgttgtggc ggatcttgaa gttggccttg
     3661 atgccgttct tctgcttgtc ggcggtgata tagacgttgt cgctaaagta gttgtactcc
     3721 agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg
     3781 ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc
     3841 ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg
     3901 tgctgcttca tgtggtcggg gtagcgggcg aagcactgca cgccccagga cagggtggtc
     3961 acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc
     4021 ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg
     4081 tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc
     4141 accatctagt atttctcctc tttctctagt agctagcata atacctagga ctgagctagc
     4201 tgtaaatact agtagcggcc gctgcaggag tcactaaggg ttagttagtt agattagcag
     4261 aaagtcaaaa gcctccgacc ggaggctttt gactaaaact tcccttgggg ttatcattgg
     4321 ggctcactca aaggcggtaa tcagataaaa aaaatcctta gctttcgcta aggatgattt
     4381 ctgctagagc tgtcagacca agtttacgag ctcgcttgga ctcctgttga tagatccagt
     4441 aatgacctca gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt
     4501 tattggtgag aatccaagca ctagggacag taagacgggt aagcctgttg atgataccgc
     4561 tgccttactg ggtgcattag ccagtctgaa tgacctgtca cgggataatc cgaagtggtc
     4621 agactggaaa atcagagggc aggaactgct gaacagcaaa aagtcagata gcaccacata
     4681 gcagacccgc cataaaacgc cctgagaagc ccgtgacggg cttttcttgt attatgggta
     4741 gtttccttgc atgaatccat aaaaggcgcc tgtagtgcca tttaccccca ttcactgcca
     4801 gagccgtgag cgcagcgaac tgaatgtcac gaaaaagaca gcgactcagg tgcctgatgg
     4861 tcggagacaa aaggaatatt cagcgatttg cccgagcttg cgagggtgct acttaagcct
     4921 ttagggtttt aaggtctgtt ttgtagagga gcaaacagcg tttgcgacat ccttttgtaa
     4981 tactgcggaa ctgactaaag tagtgagtta tacacagggc tgggatctat tctttttatc
     5041 tttttttatt ctttctttat tctataaatt ataaccactt gaatataaac aaaaaaaaca
     5101 cacaaaggtc tagcggaatt tacagagggt ctagcagaat ttacaagttt tccagcaaag
     5161 gtctagcaga atttacagat acccacaact caaaggaaaa ggacatgtaa ttatcattga
     5221 ctagcccatc tcaattggta tagtgattaa aatcac
//
