LOCUS       sequence-436939-        5710 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  tRNA reporter vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5710)
  AUTHORS   Preston MA, Porter DF, Chen F, Buter N, Lapointe CP, Keles S, Kimble
            J, Wickens M
  TITLE     Unbiased screen of RNA tailing activities reveals a poly(UG) 
            polymerase.
  JOURNAL   Nat Methods. 2019 May;16(5):437-445. doi: 10.1038/s41592-019-0370-6.
            Epub 2019 Apr 15.
  PUBMED    30988468
REFERENCE   2  (bases 1 to 5710)
  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: 
            "10.1038/s41592-019-0370-6"; journalName: "Nat Methods"; date: 
            "2019-05"; volume: "16"; issue: "5"; pages: "437-445"
COMMENT     Sequence Label: MAP80A_MS2
FEATURES             Location/Qualifiers
     source          1..5710
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..65)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        133..237
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(238..306,307..1098)
                     /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     complement(456..475)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1269..1857
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1758..1777
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2011..2028
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2145..2166
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2181..2186,2187..2204,2205..2211)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2219..2235
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2224..2246
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2243..2259
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2243..2259
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_RNA        2532..2550
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     primer_bind     complement(2804..2821)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2804..2820)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2813..2835)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3256..4602
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4713..5516)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(join(5517..5710,1..22))
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
ORIGIN
        1 aaaaaaaaaa tgatgaattg aaaagctaat tcttgaagac gaaagggcct cgtgatacgc
       61 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
      121 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
      181 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
      241 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
      301 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
      361 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
      421 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
      481 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
      541 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
      601 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
      661 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
      721 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
      781 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
      841 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
      901 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
      961 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     1021 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     1081 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     1141 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     1201 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     1261 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     1321 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1381 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     1441 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     1501 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     1561 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     1621 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     1681 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     1741 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     1801 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     1861 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     1921 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     1981 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     2041 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     2101 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     2161 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     2221 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttgca
     2281 tgcctgcagg tcgactctag aggatcccta aactaattgt tcaggcattt atacttttgg
     2341 tagttcagct agggaaggac gggttttatc tcatgttgtt cgttttgtta taaggttgtt
     2401 tcatatgtgt tttatgaacg tttaggatga cgtattgtca tactgacata tctcattttg
     2461 agatctaaca aagttcataa agaaattagg caacttggcc gagtggttaa ggcgaaagat
     2521 tagaaatctt tacatgagga tcacccatgt cgcaggttcg agtcctgcag ttgtcgtttt
     2581 ttccgaaatt tcgccttctc gagcaacttt ccaaacccga ccaagagaag gaagaaatca
     2641 ttttttttgt tgcgagttgt catagaacga ggtcgagaaa cagaagagga attgaaaatg
     2701 aattgtaaaa gcgttcgttc tatgcctcta ataagagagt acgtcttgct tgatgtgaac
     2761 ccaaatgccc acaaaaggat ccccgggtac cgagctcgaa ttcactggcc gtcgttttac
     2821 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     2881 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     2941 gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     3001 tttcacaccg cataatcgga tcgtacttgt tacccatcat tgaattttga acatccgaac
     3061 ctgggagttt tccctgaaac agatagtata tttgaacctg tataataata tatagtctag
     3121 cgctttacgg aagacaatgt atgtatttcg gttcctggag aaactattgc atctattgca
     3181 taggtaatct tgcacgtcgc atccccggtt cattttctgc gtttccatct tgcacttcaa
     3241 tagcatatct ttgttaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg
     3301 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     3361 gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca
     3421 acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat
     3481 gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa
     3541 atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt
     3601 gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga
     3661 agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc
     3721 gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt
     3781 atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga
     3841 ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta
     3901 taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca
     3961 atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta
     4021 gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga
     4081 tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc aatattttag
     4141 tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcttca gagcgctttt
     4201 ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg aacttcggaa
     4261 taggaacttc aaagcgtttc cgaaaacgag cgcttccgaa aatgcaacgc gagctgcgca
     4321 catacagctc actgttcacg tcgcacctat atctgcgtgt tgcctgtata tatatataca
     4381 tgagaagaac ggcatagtgc gtgtttatgc ttaaatgcgt acttatatgc gtctatttat
     4441 gtaggatgaa aggtagtcta gtacctcctg tgatattatc ccattccatg cggggtatcg
     4501 tatgcttcct tcagcactac cctttagctg ttctatatgc tgccactcct caattggatt
     4561 agtctcatcc ttcaatgcta tcatttcctt tgatattgga tcgatccgat gataagctgt
     4621 caaacatgag aattgggtaa taactgatat aattaaattg aagctctaat ttgtgagttt
     4681 agtatacatg catttactta taatacagtt ttttagtttt gctggccgca tcttctcaaa
     4741 tatgcttccc agcctgcttt tctgtaacgt tcaccctcta ccttagcatc ccttcccttt
     4801 gcaaatagtc ctcttccaac aataataatg tcagatcctg tagagaccac atcatccacg
     4861 gttctatact gttgacccaa tgcgtctccc ttgtcatcta aacccacacc gggtgtcata
     4921 atcaaccaat cgtaaccttc atctcttcca cccatgtctc tttgagcaat aaagccgata
     4981 acaaaatctt tgtcgctctt cgcaatgtca acagtaccct tagtatattc tccagtagat
     5041 agggagccct tgcatgacaa ttctgctaac atcaaaaggc ctctaggttc ctttgttact
     5101 tcttctgccg cctgcttcaa accgctaaca atacctgggc ccaccacacc gtgtgcattc
     5161 gtaatgtctg cccattctgc tattctgtat acacccgcag agtactgcaa tttgactgta
     5221 ttaccaatgt cagcaaattt tctgtcttcg aagagtaaaa aattgtactt ggcggataat
     5281 gcctttagcg gcttaactgt gccctccatg gaaaaatcag tcaagatatc cacatgtgtt
     5341 tttagtaaac aaattttggg acctaatgct tcaactaact ccagtaattc cttggtggta
     5401 cgaacatcca atgaagcaca caagtttgtt tgcttttcgt gcatgatatt aaatagcttg
     5461 gcagcaacag gactaggatg agtagcagca cgttccttat atgtagcttt cgacatgatt
     5521 tatcttcgtt tcctgcatgt ttttgttctg tgcagttggg ttaagaatac tgggcaattt
     5581 catgtttctt caacactaca tatgcgtata tataccaatc taagtctgtg ctccttcctt
     5641 cgttcttcct tctgttcgga gattaccgaa tcaaaaaaat ttcaaggaaa ccgaaatcaa
     5701 aaaaaagaat
//
