LOCUS       sequence-211536-        3074 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 3074)
  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 3074)
  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-25-0
FEATURES             Location/Qualifiers
     source          1..3074
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        624..642
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(925..941)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(925..941)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(938..960)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    949..965
                     /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(973..979,980..997,998..1003))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1018..1039
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1156..1173)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1327..1915)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1407..1426)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2086..2877,2878..2946))
                     /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     2709..2728
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2947..3051)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
       61 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
      121 cggagacggg cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
      181 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
      241 tactgagagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagctaacc attattatca tgacattaac ctataaaaat aggcgtatca
      361 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
      421 tcccggagac gggcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
      481 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
      541 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
      601 acaaaggatc ccccaaaatg atgtaatacg actcactata ggaaagaatt cacttaagca
      661 acaaaatgga aacaaacaaa caaacaaaca aacaaaacta gtcaacaaca acaacaacaa
      721 caacaacaac aacaacaaca acaacaacaa caacaacaac aacaacaaca acaacaacaa
      781 caacaacaac aacaacaaca acaacaacaa caacaacaac aacaacaaca acaacaacaa
      841 caacaacaac aacaacaaca acaacaacaa caacaacaaa caaacaaagt cgacctcgag
      901 ccccccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
      961 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     1021 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     1081 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     1141 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     1201 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     1261 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1321 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1381 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1441 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1501 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1561 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1621 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1681 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     1741 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     1801 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     1861 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1921 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     1981 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2041 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2101 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     2161 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     2221 accgcgagat ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     2281 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     2341 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     2401 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     2461 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     2521 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     2581 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     2641 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     2701 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     2761 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     2821 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     2881 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     2941 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3001 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     3061 ccgaaaagtg ccac
//
