LOCUS       sequence-201610-        2936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent sensor of E. coli SOS response.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2936)
  AUTHORS   Moreb EA, Hoover B, Yaseen A, Valyasevi N, Roecker Z, Menacho-Melgar
            R, Lynch MD
  TITLE     Managing the SOS Response for Enhanced CRISPR-Cas-Based 
            Recombineering in E. coli through Transient Inhibition of Host RecA 
            Activity.
  JOURNAL   ACS Synth Biol. 2017 Oct 2. doi: 10.1021/acssynbio.7b00174.
  PUBMED    28915012
REFERENCE   2  (bases 1 to 2936)
  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: "ACS Synth 
            Biol. 2017 Oct 2. doi: 10.1021/acssynbio.7b00174."
COMMENT     Sequence Label: pSMART-SOS-GFPuv
FEATURES             Location/Qualifiers
     source          1..2936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(82..795,796..798,799..801))
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(125..145)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     740..768
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     terminator      complement(996..1023)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(1039..1110)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1216..1245
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      1267..1854
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1755..1774
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1866..1893)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1914..2729)
                     /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
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2637..2656
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2730..2832)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(2833..2864)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcctatgaga gggtaatgta cttgacgtcg gagtccctgc agaaaaaaaa ccccgccctg
       61 tcaggggcgg ggtttttttt tttatttgta gagctcatcc atgccatgtg taatcccagc
      121 agcagttaca aactcaagaa ggaccatgtg gtcacgcttt tcgttgggat ctttcgaaag
      181 ggcagattgt gtcgacaggt aatggttgtc tggtaaaagg acagggccat cgccaattgg
      241 agtattttgt tgataatggt ctgctagttg aacggatcca tcttcaatgt tgtggcgaat
      301 tttgaagtta gctttgattc cattcttttg tttgtctgcc gtgatgtata cattgtgtga
      361 gttatagttg tactcgagtt tgtgtccgag aatgtttcca tcttctttaa aatcaatacc
      421 ttttaactcg atacgattaa caagggtatc accttcaaac ttgacttcag cacgcgtctt
      481 gtagttcccg tcatctttga aagatatagt gcgttcctgt acataacctt cgggcatggc
      541 actcttgaaa aagtcatgcc gtttcatatg atccggataa cgggaaaagc attgaacacc
      601 ataagagaaa gtagtgacaa gtgttggcca tggaacaggt agttttccag tagtgcaaat
      661 aaatttaagg gtaagctttc cgtatgtagc atcaccttca ccctctccac tgacagaaaa
      721 tttgtgccca ttaacatcac catctaattc aacaagaatt gggacaactc cagtgaaaag
      781 ttcttctcct ttgctagcca tgcaaacacc tctttgactt ttaaaacaat gcgttaaaaa
      841 caacaaactg tatataaaac cactggtttt atatacagtg aaaagttacg tcacctcgta
      901 aatcctgtca acccccgagg ataagagggc tgaaattgac ggtctacaat attgctcatg
      961 tgtcagcaat taacagtaaa cgaatacgct ataaacgcag aaaggcccac ccgaaggtga
     1021 gccagtgtga ctctagtaga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
     1081 gtctttcgac tgagcctttc gttttatttg atgcctggga tctacgccat ttgacacaca
     1141 acgaacgaat tcaagcttga tatcattcag gacgagcctc agactccagc gtaactggac
     1201 tgcaatcaac tcactggctc accttcacgg gtgggccttt cttcggtaga aaatcaaagg
     1261 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     1321 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaggtaact
     1381 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     1441 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1501 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     1561 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     1621 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     1681 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     1741 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     1801 tgacttgagc atcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     1861 agcaacgcag aaaggcccac ccgaaggtga gccaggtgat tacatttggg ccctcagaaa
     1921 aactcatcga gcatcaagtg aaactgcaat ttattcatat caggattatc aataccatat
     1981 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     2041 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     2101 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     2161 ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg ccagccatta
     2221 cgctcgtcat caaaatcact cgcaccaacc aaaccgttat tcattcgtga ttgcgcctga
     2281 gcgagacgaa atacgcgatc gccgttaaaa ggacaattac aaacaggaat cgaatgcaac
     2341 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     2401 aatacctgga atgctgtttt ccctgggatc gcagtggtga gtaaccatgc atcatcagga
     2461 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagcctg
     2521 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     2581 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
     2641 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctcgag
     2701 caagacgttt cccgttgaat atggctcatt ttagcttcct tagctcctga aaatctcgat
     2761 aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt
     2821 acgtgccgat caagtcaaaa gcctccggtc ggaggctttt gactttctgc tatggaggtc
     2881 aggtatgatt taaatggtca gtattgagcg atatctagag aattcgtcag aacgcc
//
