LOCUS       sequence-484246-        3755 bp    DNA     circular SYN 21-AUG-2025
DEFINITION  Medium copy mRFP1 reporter for monitoring metabolite-responsive 
            CRISPRa activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3755)
  TITLE     Metabolite-responsive scaffold RNAs for dynamic CRISPR 
            transcriptional regulation
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3755)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 21, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pColE1-MP-mRFP1
FEATURES             Location/Qualifiers
     source          1..3755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(19..607)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(99..118)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(778..1569,1570..1638))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1401..1420
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1639..1743)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1811..1829
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     RBS             2395..2406
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2413..3090
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(3603..3620)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
       61 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      121 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
      181 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
      241 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
      301 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
      361 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
      421 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
      481 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
      541 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
      601 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
      661 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
      721 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
      781 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
      841 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
      901 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
      961 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     1021 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     1081 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     1141 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     1201 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     1261 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     1321 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     1381 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     1441 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     1501 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     1561 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     1621 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     1681 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     1741 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     1801 aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattct ggcgaatcct
     1861 ctgaccagcc agaaaacgac ctttctgtgg tgaaaccgga tgctgcaatt cagagcgcca
     1921 gcaagtgggg gacagcagaa gacctgaccg ccgcagagtg gatgtttgac atggtgaaga
     1981 ctatcgcacc atcagccaga aaaccgaatt ttgctgggtg ggctaacgat atccgcctga
     2041 tgcgtgaacg tgacggacgt aaccaccgcg acatgtgtgt gctgttccgc tgggcatgcc
     2101 aggacaactt ctggtccggt aacgtgctga gcccggccaa gcttagcccg aaaagtgcca
     2161 cctgtggcaa ttccgacgtc agcatttgcg atcattcacg cagcgcttat tcagttgctc
     2221 actgcgatgt cataatcatc gctacgagct gtgaaagatg cataaagctc gtacgacgcg
     2281 ttcgctcgtc tcctcacttc tcctaactct cacacgtggc tgcaccggcc ccccccgctg
     2341 ccgcgggccg ttgacagttt cggggccgaa agggattgta cgagcgaatt cattaaagag
     2401 gagaaaggta ccatggcgag tagcgaagac gttatcaaag agttcatgcg tttcaaagtt
     2461 cgtatggaag gttccgttaa cggtcacgag ttcgaaatcg aaggtgaagg tgaaggtcgt
     2521 ccgtacgaag gtacccagac cgctaaactg aaagttacca aaggtggtcc gctgccgttc
     2581 gcttgggaca tcctgtcccc gcagttccag tacggttcca aagcttacgt taaacacccg
     2641 gctgacatcc cggactacct gaaactgtcc ttcccggaag gtttcaaatg ggaacgtgtt
     2701 atgaacttcg aagacggtgg tgttgttacc gttacccagg actcctccct gcaagacggt
     2761 gagttcatct acaaagttaa actgcgtggt accaacttcc cgtccgacgg tccggttatg
     2821 cagaaaaaaa ccatgggttg ggaagcttcc accgaacgta tgtacccgga agacggtgct
     2881 ctgaaaggtg aaatcaaaat gcgtctgaaa ctgaaagacg gtggtcacta cgacgctgaa
     2941 gttaaaacca cctacatggc taaaaaaccg gttcagctgc cgggtgctta caaaaccgac
     3001 atcaaactgg acatcacctc ccacaacgaa gactacacca tcgttgaaca gtacgaacgt
     3061 gctgaaggtc gtcactccac cggtgcttaa ggatccaaac tcgagtaagg atctgtgctt
     3121 tttttaacgc atgagaaagc ccccggaaga tcaccttccg ggggcttttt tattgcgcgg
     3181 atccttactc cgtcgacaag cttgcggccg cactcgagga attccgactc aattagttca
     3241 gtcagtttca ggatattagt catctctaca ttgattatga gtattcagaa attccttaaa
     3301 tattctgaca aatgctcttt ccctaaactc cccccataaa aaaacccgcc gaagcgggtt
     3361 tttacgttat ttgcggatta acgattactc gttatcagaa ccgcccagac ctgcgttcag
     3421 cagttctgcc aggctggcag atgcgtcttc cggaattgat ccgtcgaccg atgcccttga
     3481 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac
     3541 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctcttccgct
     3601 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3661 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3721 gcaaaaggcc agcaaaaggc caggaaccgt aaaaa
//
