LOCUS       Exported                4044 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  BIOFAB reporter plasmid for measuring rrnB T1 termination 
            efficiency.
ACCESSION   .
VERSION     .
KEYWORDS    pFAB808
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4044)
  AUTHORS   Mutalik VK, Guimaraes JC, Cambray G, Lam C, Christoffersen MJ, Mai 
            QA, Tran AB, Paull M, Keasling JD, Arkin AP, Endy D
  TITLE     Precise and reliable gene expression via standard transcription and 
            translation initiation elements.
  JOURNAL   Nat Methods. 2013 Apr;10(4):354-60. doi: 10.1038/nmeth.2404. Epub 
            2013 Mar 10.
  PUBMED    23474465
REFERENCE   2  (bases 1 to 4044)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4044
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        320..393
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    320..338
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    345..363
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             407..418
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             425..1102
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1192..1263
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             1353..1364
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1371..2084
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1401..1429)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2024..2044
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      2120..2191
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2207..2234
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(2305..2322)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2439..2984)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(2519..2538)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      3098..3192
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3223..4017)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3334..3353)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3944..3963
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 atcagatcat gatcccctgc gccatcagat ccttggcggc aagaaagcca tccagtttac
       61 tttgcagggc ttcccaacct taccagaggg cgccccagct ggcaattccg acgtctaggg
      121 ataacagggt aattacggcc ccagaattca aaagatctta agtaagtaag agtatacgta
      181 tatcggctaa aacgtattaa ggcgcttcgg cgcctttttt tatgggggta ttttcatccc
      241 aatccacacg tccaacgcac agcaaacacc acgtcgaccc tatcagctgc gtgctttcta
      301 tgagtcgttg ctgcatatct ccctatcagt gatagagatt gacatcccta tcagtgatag
      361 agatactgag cacatcagca ggacgcactg acctttggaa ttcattaaag aggagaaagg
      421 taccatggcg agtagcgaag acgttatcaa agagttcatg cgtttcaaag ttcgtatgga
      481 aggttccgtt aacggtcacg agttcgaaat cgaaggtgaa ggtgaaggtc gtccgtacga
      541 aggtacccag accgctaaac tgaaagttac caaaggtggt ccgctgccgt tcgcttggga
      601 catcctgtcc ccgcagttcc agtacggttc caaagcttac gttaaacacc cggctgacat
      661 cccggactac ctgaaactgt ccttcccgga aggtttcaaa tgggaacgtg ttatgaactt
      721 cgaagacggt ggtgttgtta ccgttaccca ggactcctcc ctgcaagacg gtgagttcat
      781 ctacaaagtt aaactgcgtg gtaccaactt cccgtccgac ggtccggtta tgcagaaaaa
      841 aaccatgggt tgggaagctt ccaccgaacg tatgtacccg gaagacggtg ctctgaaagg
      901 tgaaatcaaa atgcgtctga aactgaaaga cggtggtcac tacgacgctg aagttaaaac
      961 cacctacatg gctaaaaaac cggttcagct gccgggtgct tacaaaaccg acatcaaact
     1021 ggacatcacc tcccacaacg aagactacac catcgttgaa cagtacgaac gtgctgaagg
     1081 tcgtcactcc accggtgctt aataatctgc tgagtgatag actcaaggtc gctcctagcg
     1141 agtggccttt atgattatca ctttacttat gtgatgggaa ctgccaggca tcaaataaaa
     1201 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     1261 ctcctgggcg gactcactat agagggacaa actcaaggtc attcgcaaga gtggccttta
     1321 tgattgacct tcttccggtt aatcaattca ttaaagagga gaaaggtacc atgagcaaag
     1381 gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt gatgttaatg
     1441 ggcacaaatt ttctgtccgt ggagagggtg aaggtgatgc tacaaacgga aaactcaccc
     1501 ttaaatttat ttgcactact ggaaaactac ctgttccgtg gccaacactt gtcactactc
     1561 tgacctatgg tgttcaatgc ttttcccgtt atccggatca catgaaacgg catgactttt
     1621 tcaagagtgc catgcccgaa ggttatgtac aggaacgcac tatatctttc aaagatgacg
     1681 ggacctacaa gacgcgtgct gaagtcaagt ttgaaggtga tacccttgtt aatcgtatcg
     1741 agttaaaggg tattgatttt aaagaagatg gaaacattct tggacacaaa ctcgagtaca
     1801 actttaactc acacaatgta tacatcacgg cagacaaaca aaagaatgga atcaaagcta
     1861 acttcaaaat tcgccacaac gttgaagatg gttccgttca actagcagac cattatcaac
     1921 aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac ctgtcgacac
     1981 aatctgtcct ttcgaaagat cccaacgaaa agcgtgacca catggtcctt cttgagtttg
     2041 taactgctgc tgggattaca catggcatgg atgagctcta caaataagga tcggttgtcg
     2101 agtaaggatc tccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     2161 ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg
     2221 tgggcctttc tgcgtttata ggatcctaac tcgagctgaa gccaataggg ataacagggt
     2281 aatcctaggg atatattccg cttcctcgct cactgactcg ctacgctcgg tcgttcgact
     2341 gcggcgagcg gaaatggctt acgaacgggg cggagatttc ctggaagatg ccaggaagat
     2401 acttaacagg gaagtgagag ggccgcggca aagccgtttt tccataggct ccgcccccct
     2461 gacaagcatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac aggactataa
     2521 agataccagg cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg
     2581 tttaccggtg tcattccgct gttatggccg cgtttgtctc attccacgcc tgacactcag
     2641 ttccgggtag gcagttcgct ccaagctgga ctgtatgcac gaaccccccg ttcagtccga
     2701 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggaaagac atgcaaaagc
     2761 accactggca gcagccactg gtaattgatt tagaggagtt agtcttgaag tcatgcgccg
     2821 gttaaggcta aactgaaagg acaagttttg gtgactgcgc tcctccaagc cagttacctc
     2881 ggttcaaaga gttggtagct cagagaacct tcgaaaaacc gccctgcaag gcggtttttt
     2941 cgttttcaga gcaagagatt acgcgcagac caaaacgatc tcaagaagat catcttatta
     3001 atcagataaa atatttctag atttcagtgc aatttatctc ttcaaatgta gcacctgaag
     3061 tcagccccat acgatataag ttgttactag tgcttggatt ctcaccaata aaaaacgccc
     3121 ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta
     3181 tcaacaggag tccaagcgag ctctcgaacc ccagagtccc gctcagaaga actcgtcaag
     3241 aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa
     3301 gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc
     3361 ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt
     3421 ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc
     3481 gggcatgcgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc
     3541 gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg
     3601 atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat
     3661 tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg
     3721 ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac
     3781 agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag
     3841 ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga
     3901 cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa
     3961 tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg
     4021 aaacgatcct catcctgtct cttg
//
