LOCUS       sequence-102050-        4330 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  reporter plasmid that encodes one of five synthetic 
            sgRNA-repressible promoters that express mRFP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4330)
  AUTHORS   Nielsen AA, Voigt CA
  TITLE     Multi-input CRISPR/Cas genetic circuits that interface host 
            regulatory networks.
  JOURNAL   Mol Syst Biol. 2014 Nov 24;10:763. doi: 10.15252/msb.20145735.
  PUBMED    25422271
REFERENCE   2  (bases 1 to 4330)
  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: "Mol Syst 
            Biol."; date: "2014-11-24"; volume: "10:763. doi"; pages: " 
            10.15252/msb.20145735"
COMMENT     Sequence Label: pAN-PA4-RFP
FEATURES             Location/Qualifiers
     source          1..4330
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..792
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRTDQLEEFVHYVKGEITKVVGK"
     misc_RNA        1006..1055
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             1106..1783
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1803..1874
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1890..1917
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      2054..2148
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2640..2862
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             2910..3860
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
        1 atgagggaag cggtgatcgc cgaagtatcg actcaactat cagaggtagt tggcgtcatc
       61 gagcgccatc tcgaaccgac gttgctggcc gtacatttgt acggctccgc agtggatggc
      121 ggcctgaagc cacacagtga tattgatttg ctggttacgg tgaccgtaag gcttgatgaa
      181 acaacgcggc gagctttgat caacgacctt ttggaaactt cggcttcccc tggagagagc
      241 gagattctcc gcgctgtaga agtcaccatt gttgtgcacg acgacatcat tccgtggcgt
      301 tatccagcta agcgcgaact gcaatttgga gaatggcagc gcaatgacat tcttgcaggt
      361 atcttcgagc cagccacgat cgacattgat ctggctatct tgctgacaaa agcaagagaa
      421 catagcgttg ccttggtagg tccagcggcg gaggaactct ttgatccggt tcctgaacag
      481 gatctatttg aggcgctaaa tgaaacctta acgctatgga actcgccgcc cgactgggct
      541 ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt ggtacagcgc agtaaccggc
      601 aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat
      661 cagcccgtca tacttgaagc tagacaggct tatcttggac aagaagaaga tcgcttggcc
      721 tcgcgcacag atcagttgga agaatttgtc cactacgtga aaggcgagat caccaaggta
      781 gtcggcaaat aatactagct ccggcaaaaa aacgggcaag gtgtcaccac cctgcccttt
      841 ttctttaaaa ccgaaaagat tacttcgcgt ttgccacctg acgtctaaga actcggtacc
      901 aaaccaatta ttgaagacgc tgaaaagcgt ctttttttgt ttcggtcctc cgaatgacat
      961 gcgtctcctt tacacctcca caactagctg gtattatgct agcagctgtc accggatgtg
     1021 ctttccggtc tgatgagtcc gtgaggacga aacagcctct acaaataatt ttgtttaata
     1081 ctagagtcac acaggaaagt actagatggc ttcctccgaa gacgttatca aagagttcat
     1141 gcgtttcaaa gttcgtatgg aaggttccgt taacggtcac gagttcgaaa tcgaaggtga
     1201 aggtgaaggt cgtccgtacg aaggtaccca gaccgctaaa ctgaaagtta ccaaaggtgg
     1261 tccgctgccg ttcgcttggg acatcctgtc cccgcagttc cagtacggtt ccaaagctta
     1321 cgttaaacac ccggctgaca tcccggacta cctgaaactg tccttcccgg aaggtttcaa
     1381 atgggaacgt gttatgaact tcgaagacgg tggtgttgtt accgttaccc aggactcctc
     1441 cctgcaagac ggtgagttca tctacaaagt taaactgcgt ggtaccaact tcccgtccga
     1501 cggtccggtt atgcagaaaa aaaccatggg ttgggaagct tccaccgaac gtatgtaccc
     1561 ggaagacggt gctctgaaag gtgaaatcaa aatgcgtctg aaactgaaag acggtggtca
     1621 ctacgacgct gaagttaaaa ccacctacat ggctaaaaaa ccggttcagc tgccgggtgc
     1681 ttacaaaacc gacatcaaac tggacatcac ctcccacaac gaagactaca ccatcgttga
     1741 acagtacgaa cgtgctgaag gtcgtcactc caccggtgct taataatact agagccaggc
     1801 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
     1861 cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt
     1921 atagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     1981 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaactgtca gaccaagttt
     2041 acgagctcgc ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat
     2101 ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aagcactagg
     2161 gacagtaaga cgggtaagcc tgttgatgat accgctgcct tactgggtgc attagccagt
     2221 ctgaatgacc tgtcacggga taatccgaag tggtcagact ggaaaatcag agggcaggaa
     2281 ctgctgaaca gcaaaaagtc agatagcacc acatagcaga cccgccataa aacgccctga
     2341 gaagcccgtg acgggctttt cttgtattat gggtagtttc cttgcatgaa tccataaaag
     2401 gcgcctgtag tgccatttac ccccattcac tgccagagcc gtgagcgcag cgaactgaat
     2461 gtcacgaaaa agacagcgac tcaggtgcct gatggtcgga gacaaaagga atattcagcg
     2521 atttgcccga gcttgcgagg gtgctactta agcctttagg gttttaaggt ctgttttgta
     2581 gaggagcaaa cagcgtttgc gacatccttt tgtaatactg cggaactgac taaagtagtg
     2641 agttatacac agggctggga tctattcttt ttatcttttt ttattctttc tttattctat
     2701 aaattataac cacttgaata taaacaaaaa aaacacacaa aggtctagcg gaatttacag
     2761 agggtctagc agaatttaca agttttccag caaaggtcta gcagaattta cagataccca
     2821 caactcaaag gaaaaggaca tgtaattatc attgactagc ccatctcaat tggtatagtg
     2881 attaaaatca cctagaccaa ttgagatgta tgtctgaatt agttgttttc aaagcaaatg
     2941 aactagcgat tagtcgctat gacttaacgg agcatgaaac caagctaatt ttatgctgtg
     3001 tggcactact caaccccacg attgaaaacc ctacaaggaa agaacggacg gtatcgttca
     3061 cttataacca atacgctcag atgatgaaca tcagtaggga aaatgcttat ggtgtattag
     3121 ctaaagcaac cagagagctg atgacgagaa ctgtggaaat caggaatcct ttggttaaag
     3181 gctttgagat tttccagtgg acaaactatg ccaagttctc aagcgaaaaa ttagaattag
     3241 tttttagtga agagatattg ccttatcttt tccagttaaa aaaattcata aaatataatc
     3301 tggaacatgt taagtctttt gaaaacaaat actctatgag gatttatgag tggttattaa
     3361 aagaactaac acaaaagaaa actcacaagg caaatataga gattagcctt gatgaattta
     3421 agttcatgtt aatgcttgaa aataactacc atgagtttaa aaggcttaac caatgggttt
     3481 tgaaaccaat aagtaaagat ttaaacactt acagcaatat gaaattggtg gttgataagc
     3541 gaggccgccc gactgatacg ttgattttcc aagttgaact agatagacaa atggatctcg
     3601 taaccgaact tgagaacaac cagataaaaa tgaatggtga caaaatacca acaaccatta
     3661 catcagattc ctacctacgt aacggactaa gaaaaacact acacgatgct ttaactgcaa
     3721 aaattcagct caccagtttt gaggcaaaat ttttgagtga catgcaaagt aagcatgatc
     3781 tcaatggttc gttctcatgg ctcacgcaaa aacaacgaac cacactagag aacatactgg
     3841 ctaaatacgg aaggatctga ggttcttatg gctcttgtat ctatcagtga agcatcaaga
     3901 ctaacaaaca aaagtagaac aactgttcac cgttagatat caaagggaaa actgtccata
     3961 tgcacagatg aaaacggtgt aaaaaagata gatacatcag agcttttacg agtttttggt
     4021 gcatttaaag ctgttcacca tgaacagatc gacaatgtaa cgattatcaa aaaggatctt
     4081 cacctagatc cttttaaatt aaaaatgaag cttcgttgtg tctcaaaatc tctgatgtta
     4141 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     4201 taatacaagg ggtgtttact agaggttgat cgggcacgta agaggttcca actttcacca
     4261 taatgaaata agatcactac cgggcgtatt ttttgagtta tcgagatttt caggagctaa
     4321 ggaagctaaa
//
