LOCUS       sequence-208356-        3236 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 3236)
  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 3236)
  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: FJZ 663
FEATURES             Location/Qualifiers
     source          1..3236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(426..444)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(465..481)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(465..481)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(478..500)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    489..505
                     /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(513..519,520..537,538..543))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    558..579
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(696..713)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(867..1455)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(947..966)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1626..2417,2418..2486))
                     /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     2249..2268
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2487..2591)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2617..3072)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2754..2775)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2966..2985
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3199..3221
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3213..3230
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3214..3230
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactatagga aagaattgcc atcttggcaa ttcaacaaca acaacaacaa
       61 caacaacaac aacaacaaca acaacaacaa caacaaaaaa tgagagccaa gtggagaaag
      121 aagagaacta gaagacttaa gagaaagaga cggaaggtga gagccagatc caaataagcg
      181 gattatgagt aaataactct aattttgttt taaattcttt caagagtatc gtaatgtcat
      241 tgatgaatta acatgttagt ttctattcta cctcataatg gatctaaatt gcatactaat
      301 ctcacggtgg ggtgtaaacc attgcctact atttatatag tgctttatat atgtctcaca
      361 tagtttaatc aattgtccgt ttttttggtc gacctcgagg gggggcccgg taccagcttt
      421 tgttcccttt agtgagggtt aatttcgagc ttggcgtaat catggtcata gctgtttcct
      481 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
      541 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
      601 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
      661 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
      721 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
      781 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
      841 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
      901 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
      961 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     1021 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     1081 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     1141 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     1201 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     1261 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     1321 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     1381 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     1441 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     1501 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     1561 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     1621 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     1681 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     1741 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     1801 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     1861 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     1921 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     1981 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     2041 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     2101 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     2161 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     2221 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     2281 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     2341 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     2401 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     2461 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     2521 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     2581 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt
     2641 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     2701 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     2761 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     2821 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     2881 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     2941 agccggcgga cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     3001 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     3061 tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     3121 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     3181 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattg
//
