LOCUS       sequence-208355-        3221 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 3221)
  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 3221)
  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 617
FEATURES             Location/Qualifiers
     source          1..3221
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(411..429)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(450..466)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(450..466)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(463..485)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    474..490
                     /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(498..504,505..522,523..528))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    543..564
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(681..698)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(852..1440)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(932..951)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1611..2402,2403..2471))
                     /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     2234..2253
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2472..2576)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2602..3057)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2739..2760)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2951..2970
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3184..3206
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3198..3215
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3199..3215
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactatagga aagaattcaa caacaacaac aacaacaaca acaacaacaa
       61 caacaacaac aacaacaaca aaaaatgaga gccaagtgga gaaagaagag aactagaaga
      121 cttaagagaa agagacggaa ggtgagagcc agatccaaat aagcggatta tgagtaaata
      181 actctaattt tgttttaaat tctttcaaga gtatcgtaat gtcattgatg aattaacatg
      241 ttagtttcta ttctacctca taatggatct aaattgcata ctaatctcac ggtggggtgt
      301 aaaccattgc ctactattta tatagtgctt tatatatgtc tcacatagtt taatcaattg
      361 tccgtttttt tggtcgacct cgaggggggg cccggtacca gcttttgttc cctttagtga
      421 gggttaattt cgagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
      481 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
      541 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
      601 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
      661 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
      721 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
      781 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
      841 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
      901 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
      961 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     1021 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     1081 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     1141 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     1201 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     1261 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     1321 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     1381 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     1441 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     1501 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     1561 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     1621 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     1681 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     1741 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     1801 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     1861 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     1921 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     1981 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     2041 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     2101 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     2161 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     2221 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     2281 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     2341 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     2401 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     2461 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     2521 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     2581 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     2641 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     2701 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     2761 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     2821 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     2881 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcggacgtgg
     2941 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     3001 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     3061 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     3121 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     3181 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt g
//
