LOCUS       sequence-211622-        3009 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 3009)
  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 3009)
  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: CAA 659
FEATURES             Location/Qualifiers
     source          1..3009
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     11..28
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     12..28
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        50..68
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(421..437)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(421..437)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(434..456)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    445..461
                     /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(469..475,476..493,494..499))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    514..535
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(652..669)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(823..1411)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(903..922)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1582..2373,2374..2442))
                     /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     2205..2224
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2443..2547)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2615..2633
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2671..2693)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2793..2812
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(3006..3009,1..19)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gtcacgacgt tgtaaaacga cggccagtga attcggtaac acgcatatgt aatacgactc
       61 actataggaa acaacaacaa caacaacaac aacaacaaca acaacaacaa gaattgccat
      121 cttggcaatt ccaacaacaa caacaaaaaa tgagagaaca acaacaacaa caacaacaac
      181 aacaacaaca acaacaacaa caacaacaac aacaacaaca acaacaacaa caacaacaac
      241 aacaacaaca acaacaacaa caacaacaac aacaacaaca acaacaacaa caacaacaac
      301 aacaacaaca acaacaacaa caacaacaac aacaaatatt aagcttgttg ttgttgttgt
      361 tgttgttgaa caacaacaac aacaacaaca acaacaaata ttaagcttgg cgtaatcatg
      421 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
      481 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
      541 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
      601 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
      661 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
      721 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
      781 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
      841 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
      901 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
      961 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     1021 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     1081 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     1141 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     1201 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     1261 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     1321 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     1381 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     1441 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     1501 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     1561 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     1621 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     1681 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     1741 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     1801 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     1861 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     1921 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     1981 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     2041 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     2101 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     2161 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     2221 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     2281 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     2341 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     2401 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     2461 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     2521 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     2581 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     2641 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     2701 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     2761 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     2821 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     2881 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     2941 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     3001 gttttccca
//
