LOCUS       sequence-310112-        4431 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  J3-mRFP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4431)
  AUTHORS   Fontana J, Dong C, Kiattisewee C, Chavali VP, Tickman BI, Carothers 
            JM, Zalatan JG
  TITLE     Effective CRISPRa-mediated control of gene expression in bacteria 
            must overcome strict target site requirements.
  JOURNAL   Nat Commun. 2020 Apr 1;11(1):1618. doi: 10.1038/s41467-020-15454-y.
  PUBMED    32238808
REFERENCE   2  (bases 1 to 4431)
  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: "2020-04-1"; volume: "11(1):1618. doi"; pages: " 
            10.1038/s41467-020-15454-y"
COMMENT     Sequence Label: pJF143.J3
FEATURES             Location/Qualifiers
     source          1..4431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        177..211
                     /label=BBa_J23117
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.06"
     RBS             221..232
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             239..916
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      949..1020
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1036..1063
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1594..2544)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(3306..3400)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(3430..4221,4222..4290))
                     /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"
     primer_bind     4053..4072
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4291..4395)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcagca tttgcgatca ttcacgcagc gcttattcag ttgctcactg cgatgtcata
       61 atcatcgcta cgagctgtga aagatgcata aagctcgtac gacgcgttcg ctcgtctcct
      121 cacttctcct acggagcgtt ctggacacaa cgtcgtcttg aagttgcgat tatagattga
      181 cagctagctc agtcctaggg attgtgctag cgaattcatt aaagaggaga aaggtaccat
      241 ggcgagtagc gaagacgtta tcaaagagtt catgcgtttc aaagttcgta tggaaggttc
      301 cgttaacggt cacgagttcg aaatcgaagg tgaaggtgaa ggtcgtccgt acgaaggtac
      361 ccagaccgct aaactgaaag ttaccaaagg tggtccgctg ccgttcgctt gggacatcct
      421 gtccccgcag ttccagtacg gttccaaagc ttacgttaaa cacccggctg acatcccgga
      481 ctacctgaaa ctgtccttcc cggaaggttt caaatgggaa cgtgttatga acttcgaaga
      541 cggtggtgtt gttaccgtta cccaggactc ctccctgcaa gacggtgagt tcatctacaa
      601 agttaaactg cgtggtacca acttcccgtc cgacggtccg gttatgcaga aaaaaaccat
      661 gggttgggaa gcttccaccg aacgtatgta cccggaagac ggtgctctga aaggtgaaat
      721 caaaatgcgt ctgaaactga aagacggtgg tcactacgac gctgaagtta aaaccaccta
      781 catggctaaa aaaccggttc agctgccggg tgcttacaaa accgacatca aactggacat
      841 cacctcccac aacgaagact acaccatcgt tgaacagtac gaacgtgctg aaggtcgtca
      901 ctccaccggt gcttaaggat ccaaactcga gtaaggatct ccaggcatca aataaaacga
      961 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     1021 tactagagtc acactggctc accttcgggt gggcctttct gcgtttatac ctagggtacg
     1081 ggttttgctg cccgcaaacg ggctgttctg gtgttgctag tttgttatca gaatcgcaga
     1141 tccggcttca gccggtttgc cggctgaaag cgctatttct tccagaattg ccatgatttt
     1201 ttccccacgg gaggcgtcac tggctcccgt gttgtcggca gctttgattc gataagcagc
     1261 atcgcctgtt tcaggctgtc tatgtgtgac tgttgagctg taacaagttg tctcaggtgt
     1321 tcaatttcat gttctagttg ctttgtttta ctggtttcac ctgttctatt aggtgttaca
     1381 tgctgttcat ctgttacatt gtcgatctgt tcatggtgaa cagctttgaa tgcaccaaaa
     1441 actcgtaaaa gctctgatgt atctatcttt tttacaccgt tttcatctgt gcatatggac
     1501 agttttccct ttgatatgta acggtgaaca gttgttctac ttttgtttgt tagtcttgat
     1561 gcttcactga tagatacaag agccataaga acctcagatc cttccgtatt tagccagtat
     1621 gttctctagt gtggttcgtt gtttttgcgt gagccatgag aacgaaccat tgagatcata
     1681 cttactttgc atgtcactca aaaattttgc ctcaaaactg gtgagctgaa tttttgcagt
     1741 taaagcatcg tgtagtgttt ttcttagtcc gttatgtagg taggaatctg atgtaatggt
     1801 tgttggtatt ttgtcaccat tcatttttat ctggttgttc tcaagttcgg ttacgagatc
     1861 catttgtcta tctagttcaa cttggaaaat caacgtatca gtcgggcggc ctcgcttatc
     1921 aaccaccaat ttcatattgc tgtaagtgtt taaatcttta cttattggtt tcaaaaccca
     1981 ttggttaagc cttttaaact catggtagtt attttcaagc attaacatga acttaaattc
     2041 atcaaggcta atctctatat ttgccttgtg agttttcttt tgtgttagtt cttttaataa
     2101 ccactcataa atcctcatag agtatttgtt ttcaaaagac ttaacatgtt ccagattata
     2161 ttttatgaat ttttttaact ggaaaagata aggcaatatc tcttcactaa aaactaattc
     2221 taatttttcg cttgagaact tggcatagtt tgtccactgg aaaatctcaa agcctttaac
     2281 caaaggattc ctgatttcca cagttctcgt catcagctct ctggttgctt tagctaatac
     2341 accataagca ttttccctac tgatgttcat catctgagcg tattggttat aagtgaacga
     2401 taccgtccgt tctttccttg tagggttttc aatcgtgggg ttgagtagtg ccacacagca
     2461 taaaattagc ttggtttcat gctccgttaa gtcatagcga ctaatcgcta gttcatttgc
     2521 tttgaaaaca actaattcag acatacatct caattggtct aggtgatttt aatcactata
     2581 ccaattgaga tgggctagtc aatgataatt actagtcctt ttcccgggtg atctgggtat
     2641 ctgtaaattc tgctagacct ttgctggaaa acttgtaaat tctgctagac cctctgtaaa
     2701 ttccgctaga cctttgtgtg ttttttttgt ttatattcaa gtggttataa tttatagaat
     2761 aaagaaagaa taaaaaaaga taaaaagaat agatcccagc cctgtgtata actcactact
     2821 ttagtcagtt ccgcagtatt acaaaaggat gtcgcaaacg ctgtttgctc ctctacaaaa
     2881 cagaccttaa aaccctaaag gcttaagtag caccctcgca agctcgggca aatcgctgaa
     2941 tattcctttt gtctccgacc atcaggcacc tgagtcgctg tctttttcgt gacattcagt
     3001 tcgctgcgct cacggctctg gcagtgaatg ggggtaaatg gcactacagg cgccttttat
     3061 ggattcatgc aaggaaacta cccataatac aagaaaagcc cgtcacgggc ttctcagggc
     3121 gttttatggc gggtctgcta tgtggtgcta tctgactttt tgctgttcag cagttcctgc
     3181 cctctgattt tccagtctga ccacttcgga ttatcccgtg acaggtcatt cagactggct
     3241 aatgcaccca gtaaggcagc ggtatcatca acaggcttac ccgtcttact gtccctagtg
     3301 cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga
     3361 tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct cgtaaacttg
     3421 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3481 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3541 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3601 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3661 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3721 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     3781 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     3841 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     3901 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     3961 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4021 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4081 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4141 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4201 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4261 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4321 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4381 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gtggcaattc c
//
