LOCUS       sequence-217725-        2765 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  in vitro mRNA transcription.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2765)
  AUTHORS   Yourik P, Aitken CE, Zhou F, Gupta N, Hinnebusch AG, Lorsch JR
  TITLE     Yeast eIF4A enhances recruitment of mRNAs regardless of their 
            structural complexity.
  JOURNAL   Elife. 2017 Nov 30;6. doi: 10.7554/eLife.31476.
  PUBMED    29192585
REFERENCE   2  (bases 1 to 2765)
  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: "Elife."; 
            date: "2017-11-30"; volume: "6. doi"; pages: " 10.7554/eLife.31476"
COMMENT     Sequence Label: SL-150-0
FEATURES             Location/Qualifiers
     source          1..2765
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(299..315)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(299..315)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(312..334)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    323..339
                     /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)."
     promoter        complement(join(347..353,354..371,372..377))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    392..413
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(530..547)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(701..1289)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(781..800)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1460..2251,2252..2320))
                     /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     2083..2102
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2321..2425)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2493..2511
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2549..2571)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2671..2690
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 taatacgact cactatagga aagaattcac aacaacaaca acaacaacaa caacaacaac
       61 aacaacaaca acaacaacaa caacaactta agcaacaaca acaacaacaa caacaacaac
      121 aacaattgaa acaacaacaa caacaacaac aacaacaaca acaaaaaatg gaaacaaaca
      181 aacaaacaaa caaacaaaac tagtcaacaa caacaacaac aacaacaaca acaacaacaa
      241 caacaacaac aacaacaaca aagtcgacct cgagcccccc aagcttggcg taatcatggt
      301 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
      361 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
      421 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
      481 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
      541 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
      601 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
      661 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
      721 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
      781 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
      841 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
      901 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
      961 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     1021 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     1081 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     1141 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     1201 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     1261 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     1321 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     1381 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     1441 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     1501 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     1561 agggcttacc atctggcccc agtgctgcaa tgataccgcg agatccacgc tcaccggctc
     1621 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     1681 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     1741 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     1801 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     1861 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     1921 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     1981 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     2041 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     2101 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     2161 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     2221 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     2281 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     2341 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     2401 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     2461 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     2521 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     2581 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     2641 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     2701 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaatacaaa ggatccccca
     2761 aaatg
//
