LOCUS       sequence-178836-        4975 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 4975)
  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 4975)
  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..4975
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        11..29
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      41..50
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    840..961
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(851..870)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     905..924
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      1142..1597
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1229..1248)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1439..1460
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1664..1683)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1783..1805
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1843..1861)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1929..2033
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2034..2102,2103..2894)
                     /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(2252..2271)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3065..3653
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3554..3573
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3732..4941
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4778..4797
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
ORIGIN
        1 ctagagtact taatacgact cactataggc tagcctcgag gccaccatgg cgactgtcga
       61 accggaaacc acccctactc ctaatccccc gactacagaa gaggagaaaa cggaatctaa
      121 tcaggaggtt gctaacccag aacactatat taaacatccc ctacagaaca gatgggcact
      181 ctggtttttt aaaaatgata aaagcaaaac ttggcaagca aacctgcggc tgatctccaa
      241 gtttgatact gttgaagact tttgggctct gtacaaccat atccagttgt ctagtaattt
      301 aatgcctggc tgtgactact cactttttaa ggatggtatt gagcctatgt gggaagatga
      361 gaaaaacaaa cggggaggac gatggctaat tacattgaac aaacagcagc gaagaagtga
      421 cctcgatcgc ttttggctag agacacttct gtgccttatt ggagaatctt ttgatgacta
      481 cagtgatgat gtatgtggcg ctgttgttaa tgttagagct aaaggtgata agatagcaat
      541 atggactact gaatgtgaaa acagagaagc tgttacacat atagggaggg tgtacaagga
      601 aaggttagga cttcctccaa agatagtgat tggttatcag tcccacgcag acacagctac
      661 taagagcggc tccaccacta aaaataggtt tgttgttggg ggaggctcgg gcggtggaag
      721 tcgagagaga ccagatggca gagagtccgt ttctcattgt accgatgtcg gacggtctcc
      781 acgctaacac gcgtggtacc tctagagtcg acccgggcgg ccgcttcgag cagacatgat
      841 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
      901 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
      961 taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt
     1021 ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatccgg gctggcgtaa
     1081 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     1141 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     1201 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     1261 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     1321 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     1381 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     1441 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     1501 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     1561 aacgcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta
     1621 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
     1681 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     1741 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     1801 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     1861 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     1921 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     1981 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     2041 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     2101 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     2161 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     2221 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     2281 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     2341 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     2401 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     2461 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     2521 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     2581 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     2641 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     2701 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tcgcgcggta
     2761 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     2821 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     2881 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     2941 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     3001 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3061 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3121 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3181 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     3241 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3301 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3361 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3421 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3481 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     3541 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     3601 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     3661 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atggctcgac
     3721 agatctgatc tggcctccgc gccgggtttt ggcgcctccc gcgggcgccc ccctcctcac
     3781 ggcgagcgct gccacgtcag acgaagggcg caggagcgtc ctgatccttc cgcccggacg
     3841 ctcaggacag cggcccgctg ctcataagac tcggccttag aaccccagta tcagcagaag
     3901 gacattttag gacgggactt gggtgactct agggcactgg ttttctttcc agagagcgga
     3961 acaggcgagg aaaagtagtc ccttctcggc gattctgcgg agggatctcc gtggggcggt
     4021 gaacgccgat gattatataa ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg
     4081 ggatttgggt cgcggttctt gtttgtggat cgctgtgatc gtcacttggt gagtagcggg
     4141 ctgctgggct ggccggggct ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg
     4201 gagagtccgc caagggctgt agtctgggtc cgcgagcaag gttgccctga actgggggtt
     4261 ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg aatggaagac gcttgtgagg
     4321 cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga acccaaggtc
     4381 ttgaggcctt cgctaatgcg ggaaagctct tattcgggtg agatgggctg gggcaccatc
     4441 tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg
     4501 gggcggcagt tatgcggtgc cgttgggcag tgcacccgta cctttgggag cgcgcgccct
     4561 cgtcgtgtcg tgacgtcacc cgttctgttg gcttataatg cagggtgggg ccacctgccg
     4621 gtaggtgtgc ggtaggcttt tctccgtcgc aggacgcagg gttcgggcct agggtaggct
     4681 ctcctgaatc gacaggcgcc ggacctctgg tgaggggagg gataagtgag gcgtcagttt
     4741 ctttggtcgg ttttatgtac ctatcttctt aagtagctga agctccggtt ttgaactatg
     4801 cgctcggggt tggcgagtgt gttttgtgaa gttttttagg caccttttga aatgtaatca
     4861 tttgggtcaa tatgtaattt tcagtgttag actagtaaat tgtccgctaa attctggccg
     4921 tttttggctt ttttgttaga cgtgtccact cccagttcaa ttacagctct taagg
//
