LOCUS        pSicoR-eIF1a-GFP-RPL10a        8284 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses GFP-RPL10a for ribosomal immunoprecipitation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8284)
  AUTHORS   Rodrigues DC, Mufteev M, Weatheritt RJ, Djuric U, Ha KCH, Ross PJ, 
            Wei W, Piekna A, Sartori MA, Byres L, Mok RSF, Zaslavsky K, Pasceri 
            P, Diamandis P, Morris Q, Blencowe BJ, Ellis J
  TITLE     Shifts in Ribosome Engagement Impact Key Gene Sets in 
            Neurodevelopment and Ubiquitination in Rett Syndrome.
  JOURNAL   Cell Rep. 2020 Mar 24;30(12):4179-4196.e11. doi: 
            10.1016/j.celrep.2020.02.107.
  PUBMED    32209477
REFERENCE   2  (bases 1 to 8284)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            //gt12365.cn
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.celrep.2020.02"; journalName: "Cell Rep"; date: 
            "2020-03-24- 24"; volume: "30"; issue: "12"; pages: "4179-4196.e11"
COMMENT     Sequence Label: pSicoR-eIF1a-GFP-RPL10a
FEATURES             Location/Qualifiers
     source          1..8284
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1684..1917
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2102..2146
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2444..2561
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(2814..2847)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3007..3218
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3231..3499
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      3537..3546
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3543..3545,3546..3548,3549..4259)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3543..3545,3546..3548,3549..4259)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3588..3609)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3849..3868)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4196..4217
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(5084..5117)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5173..5761
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5226..5246)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5644..5655)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5764..5780)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5765..5781)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6290..6470
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6532..7120)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6612..6631)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7291..8082,8083..8151))
                     /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     7914..7933
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8152..8256)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga
     1561 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     1621 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     1681 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     1741 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     1801 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     1861 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     1921 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     1981 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     2041 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     2101 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     2161 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     2221 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     2281 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     2341 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact
     2401 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg
     2461 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     2521 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     2581 gcagagatcc agtttggtta gtaccgggcc cgctctagag atccgacgcg ccatctctag
     2641 gcccgcgccg gccccctcgc acagacttgt gggagaagct cggctactcc cctgccccgg
     2701 ttaatttgca tataatattt cctagtaact atagaggctt aatgtgcgat aaaagacaga
     2761 taatctgttc tttttaatac tagctacatt ttacatgata ggcttggatt tctataactt
     2821 cgtatagcat acattatacg aagttataca tgtcacaaaa ggaaactcac cctaactgta
     2881 aagtaattgt gtgttttgag actataaata tcccttggag aaaagccttg ttaacgcgcg
     2941 gtgaccctcg agtactagga tccattaggc ggccgcggat ctgcgatcgc tccggtgccc
     3001 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
     3061 attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
     3121 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
     3181 acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca
     3241 tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg
     3301 ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa
     3361 agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc
     3421 ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg
     3481 ttctgcgccg ttacagatcc aagctgtgac cggcgcctac gctagcgcta ccggtcgcca
     3541 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     3601 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     3661 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     3721 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     3781 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     3841 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     3901 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     3961 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     4021 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     4081 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     4141 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     4201 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     4261 ccggccggac tcagatctcg agctcaagct tcgaattcag cagcaaagtc tcacgcgaca
     4321 ccctgtacga ggcggtgcgg gaagtcctgc acgggaacca gcgcaagcgc cgcaagtttc
     4381 tggagacggt ggagctgcag atcagcctga agaactacga ccctcagaag gacaaacgtt
     4441 tctcgggcac cgtcaggctc aagtccaccc cacgccccaa gttctcggtg tgcgttctgg
     4501 gggaccagca gcactgtgat gaagccaagg ccgtggatat cccccacatg gacatcgagg
     4561 cgctcaagaa gcttaacaaa aacaagaagt tggtcaagaa gctggctaag aagtacgatg
     4621 cctttttggc ctctgagtct ctgattaagc agatcccacg tatcctgggc ccaggcctaa
     4681 acaaggctgg caagttcccc tccctgctga cacacaatga aaacatggtg gccaaagtgg
     4741 atgaggtgaa atcgacaatc aagttccaga tgaagaaggt gctgtgtttg gccgtcgctg
     4801 ttggccacgt gaagatgacc gatgatgagc tagtctacaa cattcatctg gctgtcaatt
     4861 tcttggtgtc cttgcttaag aaaaactggc aaaacgtgcg ggctctgtac atcaagagca
     4921 ccatgggcaa gccccagcgt ctgtattagg gatccaccgg atctagataa ctgatcataa
     4981 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     5041 tgaacctgaa acataaaatg aatgcaattc gtcgagggac ctaataactt cgtatagcat
     5101 acattatacg aagttataca tgtttaaggg ttccggttcc actaggtaca attcgatatc
     5161 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     5221 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     5281 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     5341 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     5401 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     5461 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     5521 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     5581 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     5641 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     5701 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     5761 catcgatacc gtcgacctcg atcgagacct agaaaaacat ggagcaatca caagtagcaa
     5821 tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg
     5881 ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct
     5941 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
     6001 agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt ggcagaacta
     6061 cacaccaggg ccagggatca gatatccact gacctttgga tggtgctaca agctagtacc
     6121 agttgagcaa gagaaggtag aagaagccaa tgaaggagag aacacccgct tgttacaccc
     6181 tgtgagcctg catgggatgg atgacccgga gagagaagta ttagagtgga ggtttgacag
     6241 ccgcctagca tttcatcaca tggcccgaga gctgcatccg gactgtactg ggtctctctg
     6301 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     6361 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     6421 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gcatgtgagc
     6481 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6541 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6601 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6661 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6721 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6781 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6841 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6901 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6961 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7021 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7081 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7141 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7201 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7261 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7321 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7381 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7441 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7501 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7561 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7621 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7681 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7741 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7801 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7861 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7921 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7981 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     8041 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8101 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8161 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8221 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8281 tgac
//