LOCUS       sequence-467741-        6023 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter of the SOS response.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6023)
  AUTHORS   Kaldalu N, Berzins N, Berglund Fick S, Sharma A, Andersson NC, Aedla
            J, Hinnu M, Puhar A, Hauryliuk V, Tenson T
  TITLE     Antibacterial compounds against non-growing and intracellular 
            bacteria.
  JOURNAL   NPJ Antimicrob Resist. 2025 Apr 11;3(1):25. doi: 
            10.1038/s44259-025-00097-0.
  PUBMED    40216902
REFERENCE   2  (bases 1 to 6023)
  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: "NPJ 
            Antimicrob Resist."; date: "2025-04-11"; volume: "3(1):25. doi"; 
            pages: " 10.1038/s44259-025-00097-0"
COMMENT     Sequence Label: pAED2
FEATURES             Location/Qualifiers
     source          1..6023
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     118..137
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(join(196..987,988..1056))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     819..838
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1057..1161)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(1213..1240)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        join(1448..1453,1454..1470,1471..1476)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1576..1604)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2199..2219
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             2677..3372
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(2810..2828)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3078..3097
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3475..3498
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      3583..3677
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      4169..4391
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             4439..5389
                     /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(5927..6013)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 agcttgcatg cccaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat
       61 acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt
      121 gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat
      181 gcgtccggcg tagagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
      241 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
      301 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
      361 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
      421 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
      481 taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt
      541 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
      601 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
      661 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
      721 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
      781 gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag
      841 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
      901 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
      961 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     1021 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     1081 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     1141 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     1201 cattattatc ataaaaggcc atccgtcagg atggccttct atgaatgcct tggctgcgat
     1261 tcattcttta tatgaataaa attgttgtca gtttcacgtc ttgtcctgcc atatcgcgaa
     1321 atttctgcgc aaaagcacaa aaaatttttg catctccccc ttgatgacgt gggttacgac
     1381 cccatttagt agtcaaccgc agtgagtgag tctgacgagg ccctttcgtc ttcaagaatt
     1441 ctcatgtttg acagcttatc atcgataagc tttaatgcgg tagtttatca cagttaaatt
     1501 gctaacgcag tccggggaat tcaggcttgg aggaacaata tgaaaggatc caaaggtgaa
     1561 gaattattca ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac
     1621 aaattttctg tcagtggaga gggtgaaggt gatgcaacat acggaaaact tacccttaaa
     1681 tttatttgca ctactggaaa actacctgtt ccatggccaa cacttgtcac tactttcgcg
     1741 tatggtcttc aatgctttgc gagataccca gatcatatga aacagcatga ctttttcaag
     1801 agtgccatgc ccgaaggtta tgtacaggaa agaactatat ttttcaaaga tgacgggaac
     1861 tacaagacac gtgctgaagt caagtttgaa ggtgataccc ttgttaatag aatcgagtta
     1921 aaaggtattg attttaaaga agatggaaac attcttggac acaaattgga atacaactat
     1981 aactcacaca aagtatacat catggcagac aaacaaaagg atggaatcaa agttaacttc
     2041 aaaattagac acaacattga agatggaagc gttcaactag cagaccatta tcaacgaaat
     2101 actccaattg gcgatggccc tgtcctttta ccagacaacc attacctgtc cacacaatct
     2161 gccctttcga aagatcccaa cgaaaagaga gaccacatgg tccttcttga gtttgtaaca
     2221 gctgctggga ttacacatgg tatggatgaa ttgtacaaat aaccgccacc ccctgaacct
     2281 gaaacataaa atgaatagaa atcatcctta gcgaaagcta aggatttttt ttgagctcgg
     2341 cgcgcctgca ggtcgacaag cttggaaatg acctgacagc gctcttcggc ttcggtcagt
     2401 ctatggcgtt tcatggtggt ctccgggtgt ttctctggtg tcataacagt ttaactgttg
     2461 gctctctcct tatgctaatc agccttcttt catgcctgtt gccattgagt ttaagaatga
     2521 ttggttgttt ttcttgagcg ttttcacaaa aacgtaaatc cagtgggttg acaggattta
     2581 cgaggtgacg taacttttca ctgtatataa aaccagtggt tttatataca gtttgttgtt
     2641 tttaacgcat tgttttaaaa gtcaaggagg tgcaatatgg tgagcaaggg cgaggcagtg
     2701 atcaaggagt tcatgcggtt caaggtgcac atggagggct ccatgaacgg ccacgagttc
     2761 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
     2821 gtgaccaagg gtggccccct gcccttctcc tgggacatcc tgtcccctca gttcatgtac
     2881 ggctccaggg ccttcatcaa gcaccccgcc gacatccccg actactataa gcagtccttc
     2941 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgc cgtgaccgtg
     3001 acccaggaca cctccctgga ggacggcacc ctgatctaca aggtgaagct ccgcggcacc
     3061 aacttccctc ctgacggccc cgtaatgcag aagaagacaa tgggctggga agcgtccacc
     3121 gagcggttgt accccgagga cggcgtgctg aagggcgaca ttaagatggc cctgcgcctg
     3181 aaggacggcg gccgctacct ggcggacttc aagaccacct acaaggccaa gaagcccgtg
     3241 cagatgcccg gcgcctacaa cgtcgaccgc aagttggaca tcacctccca caacgaggac
     3301 tacaccgtgg tggaacagta cgaacgctcc gagggccgcc actccaccgg cggcatggac
     3361 gagctgtaca agtgaccgcc taatgagcgg gctttaatca ctagtgaatt cgcggccgcc
     3421 tgcaggtcga ccatatggga gagctccgtc gacaagcttg cggccgcact cgagcaccac
     3481 caccaccacc accaccacta attgattaat acctaggctg ctaaacaaag cccatggatc
     3541 gatagctggt cgacaggact ctggggttcg agagctcgct tggactcctg ttgatagatc
     3601 cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc cgccgggcgt
     3661 tttttattgg tgagaatcca agcactaggg acagtaagac gggtaagcct gttgatgata
     3721 ccgctgcctt actgggtgca ttagccagtc tgaatgacct gtcacgggat aatccgaagt
     3781 ggtcagactg gaaaatcaga gggcaggaac tgctgaacag caaaaagtca gatagcacca
     3841 catagcagac ccgccataaa acgccctgag aagcccgtga cgggcttttc ttgtattatg
     3901 ggtagtttcc ttgcatgaat ccataaaagg cgcctgtagt gccatttacc cccattcact
     3961 gccagagccg tgagcgcagc gaactgaatg tcacgaaaaa gacagcgact caggtgcctg
     4021 atggtcggag acaaaaggaa tattcagcga tttgcccgag cttgcgaggg tgctacttaa
     4081 gcctttaggg ttttaaggtc tgttttgtag aggagcaaac agcgtttgcg acatcctttt
     4141 gtaatactgc ggaactgact aaagtagtga gttatacaca gggctgggat ctattctttt
     4201 tatctttttt tattctttct ttattctata aattataacc acttgaatat aaacaaaaaa
     4261 aacacacaaa ggtctagcgg aatttacaga gggtctagca gaatttacaa gttttccagc
     4321 aaaggtctag cagaatttac agatacccac aactcaaagg aaaaggacta gtaattatca
     4381 ttgactagcc catctcaatt ggtatagtga ttaaaatcac ctagaccaat tgagatgtat
     4441 gtctgaatta gttgttttca aagcaaatga actagcgatt agtcgctatg acttaacgga
     4501 gcatgaaacc aagctaattt tatgctgtgt ggcactactc aaccccacga ttgaaaaccc
     4561 tacaaggaaa gaacggacgg tatcgttcac ttataaccaa tacgctcaga tgatgaacat
     4621 cagtagggaa aatgcttatg gtgtattagc taaagcaacc agagagctga tgacgagaac
     4681 tgtggaaatc aggaatcctt tggttaaagg ctttgagatt ttccagtgga caaactatgc
     4741 caagttctca agcgaaaaat tagaattagt ttttagtgaa gagatattgc cttatctttt
     4801 ccagttaaaa aaattcataa aatataatct ggaacatgtt aagtcttttg aaaacaaata
     4861 ctctatgagg atttatgagt ggttattaaa agaactaaca caaaagaaaa ctcacaaggc
     4921 aaatatagag attagccttg atgaatttaa gttcatgtta atgcttgaaa ataactacca
     4981 tgagtttaaa aggcttaacc aatgggtttt gaaaccaata agtaaagatt taaacactta
     5041 cagcaatatg aaattggtgg ttgataagcg aggccgcccg actgatacgt tgattttcca
     5101 agttgaacta gatagacaaa tggatctcgt aaccgaactt gagaacaacc agataaaaat
     5161 gaatggtgac aaaataccaa caaccattac atcagattcc tacctacata acggactaag
     5221 aaaaacacta cacgatgctt taactgcaaa aattcagctc accagttttg aggcaaaatt
     5281 tttgagtgac atgcaaagta agtatgatct caatggttcg ttctcatggc tcacgcaaaa
     5341 acaacgaacc acactagaga acatactggc taaatacgga aggatctgag gttcttatgg
     5401 ctcttgtatc tatcagtgaa gcatcaagac taacaaacaa aagtagaaca actgttcacc
     5461 gttacatatc aaagggaaaa ctgtccatat gcacagatga aaacggtgta aaaaagatag
     5521 atacatcaga gcttttacga gtttttggtg cattcaaagc tgttcaccat gaacagatcg
     5581 acaatgtaac agatgaacag catgtaacac ctaatagaac aggtgaaacc agtaaaacaa
     5641 agcaactaga acatgaaatt gaacacctga gacaacttgt tacagctcaa cagtcacaca
     5701 tagacagcct gaaacaggcg atgctgctta tcgaatcaaa gctgccgaca acacgggagc
     5761 cagtgacgcc tcccgtgggg aaaaaatcat ggcaattctg gaagaaatag cgctttcagc
     5821 cggcaaaccg gctgaagccg gatctgcgat tctgataaca aactagcaac accagaacag
     5881 cccgtttgcg ggcagcaaaa cccgtaccct aggtctaggg cggcggattt gtcctactca
     5941 ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtctttc gactgagcct
     6001 ttcgttttat ttgatgcctc tag
//
