LOCUS       sequence-106571-        2800 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This pFTV1 vector contains a no-aptamer control mRNA encoding mRFP1 
            gene. It should be used when testing AEB-F12 and AEB-Theo45 
            riboswitches to measure their non-specific ligand effect on mRFP1 
            expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 66 to 100)
  AUTHORS   Espah Borujeni A, Mishler DM, Wang J, Huso W, Salis HM
  TITLE     Automated physics-based design of synthetic riboswitches from 
            diverse RNA aptamers.
  JOURNAL   Nucleic Acids Res. 2015 Nov 30. pii: gkv1289.
  PUBMED    26621913
REFERENCE   2  (bases 1 to 2800)
  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: "Nucleic 
            Acids Res. 2015 Nov 30. pii: gkv1289."
COMMENT     Sequence Label: AEB-No-aptamer-controlmRFP1
FEATURES             Location/Qualifiers
     source          1..2800
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        66..100
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     RBS             116..124
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             130..807
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      852..923
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      939..966
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      1008..1094
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(1259..1847)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1339..1358)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1936..2030)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2051..2710)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2625..2644
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(2711..2800,1..13))
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 tacgtgccga tcaacgtctc attttcgcca gatatcctcg agcgcggaat tccctagggg
       61 atccgttgac ggctagctca gtcctaggta cagtgctagc ttctagaaaa aaaaataagg
      121 aggtaaaaaa tggcgagctc tgaagacgtt atcaaagagt tcatgcgttt caaagttcgt
      181 atggaaggtt ccgttaacgg tcacgagttc gaaatcgaag gtgaaggtga aggtcgtccg
      241 tacgaaggta cccagaccgc taaactgaaa gttaccaaag gtggtccgct gccgttcgct
      301 tgggacatcc tgtccccgca gttccagtac ggttccaaag cttacgttaa acacccggct
      361 gacatcccgg actacctgaa actgtccttc ccggaaggtt tcaaatggga acgtgttatg
      421 aacttcgaag acggtggtgt tgttaccgtt acccaggact cctccctgca agacggtgag
      481 ttcatctaca aagttaaact gcgtggtacc aacttcccgt ccgacggtcc ggttatgcag
      541 aaaaaaacca tgggttggga agcttccacc gaacgtatgt acccggaaga cggtgctctg
      601 aaaggtgaaa tcaaaatgcg tctgaaactg aaagacggtg gtcactacga cgctgaagtt
      661 aaaaccacct acatggctaa aaaaccggtt cagctgccgg gtgcttacaa aaccgacatc
      721 aaactggaca tcacctccca caacgaagac tacaccatcg ttgaacagta cgaacgtgct
      781 gaaggtcgtc actccaccgg tgcttaataa ggccggccgt gctagtgtag atcgctacta
      841 gagccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
      901 gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg ggtgggcctt
      961 tctgcgttta tacaattgac tagtctgcag aaaagcggcc gcggcatcaa ataaaacgaa
     1021 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     1081 tgagtaggac aaatccgccg ccctagagca tgcgcgttcg gctgcggcga gcggtatcag
     1141 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1201 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1261 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1321 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1381 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1441 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     1501 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     1561 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     1621 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     1681 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     1741 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     1801 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     1861 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     1921 tgcccgggtg cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa
     1981 caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc caagcggtac
     2041 cgatatcaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca
     2101 ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc agcggcatca
     2161 gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt
     2221 ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgagacga
     2281 aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca
     2341 catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg
     2401 atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata
     2461 tcaccagctc accgtctttc attgccatac gaaattccgg atgagcattc atcaggcggg
     2521 caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa
     2581 aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg
     2641 cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt
     2701 ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg
     2761 gtagtgatct tatttcatta tggtgaaagt tggaacctct
//
