LOCUS       sequence-106559-        4242 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid is designed to validate the biophysical model of 
            translational coupling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 135 to 169)
  AUTHORS   Tian T, Salis HM
  TITLE     A predictive biophysical model of translational coupling to 
            coordinate and control protein expression in bacterial operons.
  JOURNAL   Nucleic Acids Res. 2015 Aug 18;43(14):7137-51. doi: 
            10.1093/nar/gkv635. Epub 2015 Jun 27.
  PUBMED    26117546
REFERENCE   2  (bases 1 to 4242)
  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"; doi: 
            "10.1093/nar/gkv635"; journalName: "Nucleic Acids Res"; date: 
            "2015-08-18- 18"; volume: "43"; issue: "14"; pages: "7137-51"
COMMENT     Sequence Label: pTd-4-15-13-1
FEATURES             Location/Qualifiers
     source          1..4242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      39..66
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        135..169
                     /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             947..955
                     /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             959..1636
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MKASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIK
                     MRLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRH
                     STGA"
     RBS             1650..1658
                     /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)"
     terminator      2455..2541
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(2706..3294)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2786..2805)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3383..3477)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3498..4157)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4072..4091
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 cctcttacgt gaacctaggc cttactgcag atcacactgg ctcaccttcg ggtgggcctt
       61 tctgcgttta tatactagag agagaatata aaaagccaga ttattaatcc ggctttttta
      121 ttatttagga tccattgacg gctagctcag tccaaggtac agtgctagct tctagacgaa
      181 gaccatgatc acaacttaat gacgtattca tggtcgacaa gggcgaagaa ctctttaccg
      241 gcgtcgtccc gatcctggta gagcttgacg gcgatgtcaa cggtcataag tttagcgtta
      301 gcggcgaagg cgagggcgac gcgacgtacg gtaaattaac cttaaaattc atctgtacaa
      361 ccggtaaatt accggtaccg tggcccacgt tagttaccac ccttacctgg ggcgtccagt
      421 gcttcgcgcg ctatccagac cacatgaagc aacacgattt cttcaaatcg gctatgcccg
      481 aaggctatgt acaggagcgc accatcttct tcaaggacga cggtaattat aaaacccgcg
      541 cagaagttaa gttcgagggc gatacgctcg ttaatcgcat cgaactcaaa ggtatcgact
      601 tcaaagagga cggcaacatt ttaggtcaca agttagagta taacgcgatt tccgataacg
      661 tctacattac cgccgataaa caaaagaacg gtatcaaagc aaactttaag attcgtcata
      721 acatcgagga cggctcggta cagctcgccg accattacca acagaatacg ccgatcggcg
      781 acggtccggt cctcctcccg gacaatcatt atctcagcac ccagagcgcg ctctccaaag
      841 atccaaacga gaagcgcgac cacatggtcc ttctcgaatt tgtaacggcg gcaggcatca
      901 ccctcggtat ggacgaactc tataaggacg tcaaaaaata cctccgtaag gaggtatcat
      961 gaaagcgagc tccgaagacg ttatcaaaga atttatgcgt tttaaagttc gcatggaggg
     1021 ctcggttaac ggtcacgagt tcgaaatcga aggggaaggc gaaggtcgtc cgtacgaagg
     1081 tacccagacc gctaaactca aagttaccaa aggcggtccc ttaccgttcg cttgggatat
     1141 tttaagcccg cagttccagt acggttccaa agcttacgtt aaacacccgg cggacatccc
     1201 ggactacctt aaattatcct tccccgaagg gtttaagtgg gagcgcgtta tgaacttcga
     1261 ggacggcggc gtcgttaccg ttacccagga ctcctccctt caagacggcg agttcatcta
     1321 caaagttaaa cttcgcggta ccaacttccc atccgatggt ccggtcatgc agaagaagac
     1381 catgggttgg gaagcttcca ccgagcgcat gtatccggag gacggcgctt taaaaggcga
     1441 aatcaagatg cgcttaaagt taaaagacgg cggtcactac gacgcagaag ttaaaacgac
     1501 gtacatggcc aagaaaccgg ttcagttacc gggcgcttac aaaaccgaca tcaaactcga
     1561 catcaccagc cacaacgaag actacaccat cgtcgaacag tacgaacgcg cagaaggtcg
     1621 tcacagcacc ggtgctgaat tcgtacacct aaggaggtgt acagcatgaa acgtaaactc
     1681 gaggaacttt tcacgggggt cgtcccaatt ctggtagaat tagacggtga tgtaaacggg
     1741 cacaagttta gcgtcagcgg agagggcgag ggggacgcca cctacggtaa acttaccctt
     1801 aagtttattt gcacgacggg caaattaccc gttccgtggc caacgctcgt cactactttc
     1861 ggctacggcg ttcagtgctt cgcgagatat ccggatcaca tgaagcagca cgatttcttc
     1921 aagtcggcca tgccagaggg ttacgttcag gagagaacta tatttttcaa ggacgacggc
     1981 aattacaaga cacgcgccga agtcaagttc gaaggcgata ccctggttaa ccgtatcgag
     2041 ttaaaaggta tcgattttaa agaagacgga aacattctcg gtcacaaact cgaatacaac
     2101 tataacagcc ataatgtcta cattatggct gacaaacaga agaacggcat taaggttaac
     2161 ttcaaaattc gccacaacat cgaagacggt agcgtccaac tagcagacca ttatcaacaa
     2221 aatactccta tcggcgacgg cccagtcctt ttaccagaca accattacct tagcacacaa
     2281 agcgccctta gcaaagatcc caacgagaag cgcgaccaca tggtccttct cgaatttgtc
     2341 acagcggcag ggatcaccca cggcatggac gaactctaca aataataatt aattaaaaaa
     2401 aaaaaccccg cccctgacag ggcggggttt ttttttctaa tatgcggccg ccatcaaata
     2461 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     2521 gctctcctga gtaggacaaa tccgccgccc tagagcatgc gcgttcggct gcggcgagcg
     2581 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2641 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2701 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2761 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2821 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2881 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2941 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3001 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3061 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3121 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     3181 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3241 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3301 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3361 ttggtcatgc ccgggtgctt ggattctcac caataaaaaa cgcccggcgg caaccgagcg
     3421 ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa
     3481 gcggtaccga tatcaaatta cgccccgccc tgccactcat cgcagtactg ttgtaattca
     3541 ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc
     3601 ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag
     3661 aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca gggattggct
     3721 gagacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa
     3781 cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc
     3841 cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta
     3901 tcccatatca ccagctcacc gtctttcatt gccatacgaa attccggatg agcattcatc
     3961 aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc
     4021 tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac
     4081 tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca
     4141 gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat
     4201 acgcccggta gtgatcttat ttcattatgg tgaaagttgg aa
//
