LOCUS       sequence-197845-        4980 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EIF4e plasmid with Pum cloning site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4980)
  AUTHORS   Adamala KP, Martin-Alarcon DA, Boyden ES
  TITLE     Programmable RNA-binding protein composed of repeats of a single 
            modular unit.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 May 10;113(19):E2579-88. doi: 
            10.1073/pnas.1519368113. Epub 2016 Apr 26.
  PUBMED    27118836
REFERENCE   2  (bases 1 to 4980)
  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"; doi: 
            "10.1073/pnas.1519368113"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2016-05-10- 10"; volume: "113"; issue: "19"; pages: 
            "E2579-88"
COMMENT     Sequence Label: [UBC][EIF4e][GG cloning]
FEATURES             Location/Qualifiers
     source          1..4980
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        12..30
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      42..51
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    841..962
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(852..871)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     906..925
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      1143..1598
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1230..1249)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1440..1461
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1665..1684)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1784..1806
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1844..1862)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1930..2034
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2035..2103,2104..2895)
                     /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     complement(2253..2272)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3066..3654
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3555..3574
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3736..4947
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4784..4803
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
ORIGIN
        1 gctagagtac ttaatacgac tcactatagg ctagcctcga ggccaccatg gcgactgtcg
       61 aaccggaaac cacccctact cctaatcccc cgactacaga agaggagaaa acggaatcta
      121 atcaggaggt tgctaaccca gaacactata ttaaacatcc cctacagaac agatgggcac
      181 tctggttttt taaaaatgat aaaagcaaaa cttggcaagc aaacctgcgg ctgatctcca
      241 agtttgatac tgttgaagac ttttgggctc tgtacaacca tatccagttg tctagtaatt
      301 taatgcctgg ctgtgactac tcacttttta aggatggtat tgagcctatg tgggaagatg
      361 agaaaaacaa acggggagga cgatggctaa ttacattgaa caaacagcag cgaagaagtg
      421 acctcgatcg cttttggcta gagacacttc tgtgccttat tggagaatct tttgatgact
      481 acagtgatga tgtatgtggc gctgttgtta atgttagagc taaaggtgat aagatagcaa
      541 tatggactac tgaatgtgaa aacagagaag ctgttacaca tatagggagg gtgtacaagg
      601 aaaggttagg acttcctcca aagatagtga ttggttatca gtcccacgca gacacagcta
      661 ctaagagcgg ctccaccact aaaaataggt ttgttgttgg gggaggctcg ggcggtggaa
      721 gtcgagagag accagatggc agagagtccg tttctcattg taccgatgtc ggacggtctc
      781 cacgctaaca cgcgtggtac ctctagagtc gacccgggcg gccgcttcga gcagacatga
      841 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
      901 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
      961 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
     1021 tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatccg ggctggcgta
     1081 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     1141 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     1201 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     1261 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     1321 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     1381 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     1441 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     1501 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     1561 taacgcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt
     1621 acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat
     1681 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     1741 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     1801 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     1861 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     1921 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     1981 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     2041 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     2101 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     2161 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     2221 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     2281 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     2341 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     2401 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     2461 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     2521 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     2581 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     2641 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     2701 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtcgcgcggt
     2761 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     2821 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     2881 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     2941 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     3001 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3061 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3121 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3181 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     3241 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3301 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3361 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3421 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3481 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3541 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3601 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3661 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catggctcga
     3721 cagatctcca gatctggcct ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcc
     3781 tcacggcgag cgctgccacg tcagacgaag ggcgcagcga gcgtcctgat ccttccgccc
     3841 ggacgctcag gacagcggcc cgctgctcat aagactcggc cttagaaccc cagtatcagc
     3901 agaaggacat tttaggacgg gacttgggtg actctagggc actggttttc tttccagaga
     3961 gcggaacagg cgaggaaaag tagtcccttc tcggcgattc tgcggaggga tctccgtggg
     4021 gcggtgaacg ccgatgatta tataaggacg cgccgggtgt ggcacagcta gttccgtcgc
     4081 agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta
     4141 gcgggctgct gggctggccg gggctttcgt ggccgccggg ccgctcggtg ggacggaagc
     4201 gtgtggagag tccgccaagg gctgtagtct gggtccgcga gcaaggttgc cctgaactgg
     4261 gggttggggg gagcgcagca aaatggcggc tgttcccgag tcttgaatgg aagacgcttg
     4321 tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg caagaaccca
     4381 aggtcttgag gccttcgcta atgcgggaaa gctcttattc gggtgagatg ggctggggca
     4441 ccatctgggg accctgacgt gaagtttgtc actgactgga gaactcggtt tgtcgtctgt
     4501 tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca cccgtacctt tgggagcgcg
     4561 cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg gtggggccac
     4621 ctgccggtag gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc gggcctaggg
     4681 taggctctcc tgaatcgaca ggcgccggac ctctggtgag gggagggata agtgaggcgt
     4741 cagtttcttt ggtcggtttt atgtacctat cttcttaagt agctgaagct ccggttttga
     4801 actatgcgct cggggttggc gagtgtgttt tgtgaagttt tttaggcacc ttttgaaatg
     4861 taatcatttg ggtcaatatg taattttcag tgttagacta gtaaattgtc cgctaaattc
     4921 tggccgtttt tggctttttt gttagacgtg tccactccca gttcaattac agctcttaag
//
