LOCUS       sequence-64680-e        5047 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter plasmid for measuring terminator strength by inserting 
            terminator of interest between the GFP and RFP, expression driven by
            a single PBAD promoter inducible by arabinose.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5047)
  AUTHORS   Chen YJ, Liu P, Nielsen AA, Brophy JA, Clancy K, Peterson T, Voigt 
            CA
  TITLE     Characterization of 582 natural and synthetic terminators and 
            quantification of their design constraints.
  JOURNAL   Nat Methods. 2013 Jun 2. doi: 10.1038/nmeth.2515.
  PUBMED    23727987
REFERENCE   2  (bases 1 to 5047)
  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: "Nat 
            Methods. 2013 Jun 2. doi: 10.1038/nmeth.2515."
COMMENT     Sequence Label: pGR
FEATURES             Location/Qualifiers
     source          1..5047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             27..38
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             45..722
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      767..838
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      854..881
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(975..992)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1146..1734)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1226..1245)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1905..2696,2697..2765))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2528..2547
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2766..2870)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2938..2956
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2960..3003)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             complement(3092..3970)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3997..4281
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     4204..4223
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             4310..4321
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4328..5041
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(4358..4386)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4981..5001
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
ORIGIN
        1 gaattcacta gtagcggccg ctgcagaaag aggagaaata ctagatggct tcctccgaag
       61 acgttatcaa agagttcatg cgtttcaaag ttcgtatgga aggttccgtt aacggtcacg
      121 agttcgaaat cgaaggtgaa ggtgaaggtc gtccgtacga aggtacccag accgctaaac
      181 tgaaagttac caaaggtggt ccgctgccgt tcgcttggga catcctgtcc ccgcagttcc
      241 agtacggttc caaagcttac gttaaacacc cggctgacat cccggactac ctgaaactgt
      301 ccttcccgga aggtttcaaa tgggaacgtg ttatgaactt cgaagacggt ggtgttgtta
      361 ccgttaccca ggactcctcc ctgcaagacg gtgagttcat ctacaaagtt aaactgcgtg
      421 gtaccaactt cccgtccgac ggtccggtta tgcagaaaaa aaccatgggt tgggaagctt
      481 ccaccgaacg tatgtacccg gaagacggtg ctctgaaagg tgaaatcaaa atgcgtctga
      541 aactgaaaga cggtggtcac tacgacgctg aagttaaaac cacctacatg gctaaaaaac
      601 cggttcagct gccgggtgct tacaaaaccg acatcaaact ggacatcacc tcccacaacg
      661 aagactacac catcgttgaa cagtacgaac gtgctgaagg tcgtcactcc accggtgctt
      721 aataacgctg atagtgctag tgtagatcgc tactagagcc aggcatcaaa taaaacgaaa
      781 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcta
      841 ctagagtcac actggctcac cttcgggtgg gcctttctgc gtttatatac tagagtcaat
      901 tgaggtagag gtacacacgc gaactccgat agccaattca gagtaataaa ctgtgataat
      961 caatgcagtg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     1021 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     1081 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     1141 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     1201 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     1261 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     1321 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     1381 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     1441 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     1501 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     1561 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     1621 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     1681 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     1741 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     1801 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     1861 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     1921 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     1981 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     2041 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     2101 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     2161 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     2221 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     2281 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     2341 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     2401 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     2461 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     2521 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     2581 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     2641 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     2701 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     2761 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     2821 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     2881 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     2941 aggcgtatca cgaggcagaa tttcagataa aaaaaatcct tagctttcgc taaggatgat
     3001 ttctggaatt gaattgaggt agaggtacac acgcgaactc cgatagccaa ttcagagtaa
     3061 taaactgtga taatcaatgc atgtactaga gttatgacaa cttgacggct acatcattca
     3121 ctttttcttc acaaccggca cggaactcgc tcgggctggc cccggtgcat tttttaaata
     3181 cccgcgagaa atagagttga tcgtcaaaac caacattgcg accgacggtg gcgataggca
     3241 tccgggtggt gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg cgccagctta
     3301 agacgctaat ccctaactgc tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa
     3361 catgctgtgc gacgctggcg atatcaaaat tgctgtctgc caggtgatcg ctgatgtact
     3421 gacaagcctc gcgtacccga ttatccatcg gtggatggag cgactcgtta atcgcttcca
     3481 tgcgccgcag taacaattgc tcaagcagat ttatcgccag cagctccgaa tagcgccctt
     3541 ccccttgccc ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt
     3601 catccgggcg aaagaacccc gtattggcaa atattgacgg ccagttaagc cattcatgcc
     3661 agtaggcgcg cggacgaaag taaacccact ggtgatacca ttcgcgagcc tccggatgac
     3721 gaccgtagtg atgaatctct cctggcggga acagcaaaat atcacccggt cggcaaacaa
     3781 attctcgtcc ctgatttttc accaccccct gaccgcgaat ggtgagattg agaatataac
     3841 ctttcattcc cagcggtcgg tcgataaaaa aatcgagata accgttggcc tcaatcggcg
     3901 ttaaacccgc caccagatgg gcattaaacg agtatcccgg cagcagggga tcattttgcg
     3961 cttcagccat acttttcata ctcccgccat tcagagaaga aaccaattgt ccatattgca
     4021 tcagacattg ccgtcactgc gtcttttact ggctcttctc gctaaccaaa ccggtaaccc
     4081 cgcttattaa aagcattctg taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca
     4141 aaagtgtcta taatcacggc agaaaagtcc acattgatta tttgcacggc gtcacacttt
     4201 gctatgccat agcattttta tccataagat tagcggatcc tacctgacgc tttttatcgc
     4261 aactctctac tgtttctcca tacccgtttt tttgggctag ctactagaga aagaggagaa
     4321 atactagatg cgtaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt
     4381 agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgcaac
     4441 atacggaaaa cttaccctta aatttatttg cactactgga aaactacctg ttccatggcc
     4501 aacacttgtc actactttcg gttatggtgt tcaatgcttt gcgagatacc cagatcatat
     4561 gaaacagcat gactttttca agagtgccat gcccgaaggt tatgtacagg aaagaactat
     4621 atttttcaaa gatgacggga actacaagac acgtgctgaa gtcaagtttg aaggtgatac
     4681 ccttgttaat agaatcgagt taaaaggtat tgattttaaa gaagatggaa acattcttgg
     4741 acacaaattg gaatacaact ataactcaca caatgtatac atcatggcag acaaacaaaa
     4801 gaatggaatc aaagttaact tcaaaattag acacaacatt gaagatggaa gcgttcaact
     4861 agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa
     4921 ccattacctg tccacacaat ctgccctttc gaaagatccc aacgaaaaga gagaccacat
     4981 ggtccttctt gagtttgtaa cagctgctgg gattacacat ggcatggatg aactatacaa
     5041 ataataa
//
