LOCUS       sequence-464902-        6349 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of reporter gene with 7xCGCGCG motifs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6349)
  AUTHORS   Saito H, Handa Y, Chen M, Schneider-Poetsch T, Shichino Y, Takahashi
            M, Romo D, Yoshida M, Furstner A, Ito T, Fukuzawa K, Iwasaki S
  TITLE     DMDA-PatA mediates RNA sequence-selective translation repression by 
            anchoring eIF4A and DDX3 to GNG motifs.
  JOURNAL   Nat Commun. 2024 Sep 2;15(1):7418. doi: 10.1038/s41467-024-51635-9.
  PUBMED    39223140
REFERENCE   2  (bases 1 to 6349)
  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: "Nat 
            Commun."; date: "2024-09-2"; volume: "15(1):7418. doi"; pages: " 
            10.1038/s41467-024-51635-9"
COMMENT     Sequence Label: psiCHECK2-7xCGCGCG
FEATURES             Location/Qualifiers
     source          1..6349
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..419
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        62..253
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(85..105)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      270..405
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          489..621
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     604..623
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     666..685
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        666..684
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             770..1705
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    1764..1812
                     /note="synthetic polyadenylation signal"
     promoter        1821..2572
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      2602..2611
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2608..4260
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /db_xref="GI:42718124"
                     /protein_id="AAS38485.1"
                     /translation="MADAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     RBS             3567..3575
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    4304..4425
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4315..4334)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4369..4388
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4558..4662
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4663..4731,4732..5523)
                     /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(4881..4900)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5694..6282
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6183..6202
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct
       61 tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
      121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
      181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
      241 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
      301 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
      361 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
      421 agcttgattc ttctgacaca acagtctcga acttaagctg cagaagttgg tcgtgaggca
      481 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
      541 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
      601 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga
      661 gtacttaata cgactcacta tagggaacgc gcgcaacaac gcgcgcaaca acgcgcgcaa
      721 caacgcgcgc aacaacgcgc gcaacaacgc gcgcaacaac gcgcgcacca tggcttccaa
      781 ggtgtacgac cccgagcaac gcaaacgcat gatcactggg cctcagtggt gggctcgctg
      841 caagcaaatg aacgtgctgg actccttcat caactactat gattccgaga agcacgccga
      901 gaacgccgtg atttttctgc atggtaacgc tgcctccagc tacctgtgga ggcacgtcgt
      961 gcctcacatc gagcccgtgg ctagatgcat catccctgat ctgatcggaa tgggtaagtc
     1021 cggcaagagc gggaatggct catatcgcct cctggatcac tacaagtacc tcaccgcttg
     1081 gttcgagctg ctgaaccttc caaagaaaat catctttgtg ggccacgact ggggggcttg
     1141 tctggccttt cactactcct acgagcacca agacaagatc aaggccatcg tccatgctga
     1201 gagtgtcgtg gacgtgatcg agtcctggga cgagtggcct gacatcgagg aggatatcgc
     1261 cctgatcaag agcgaagagg gcgagaaaat ggtgcttgag aataacttct tcgtcgagac
     1321 catgctccca agcaagatca tgcggaaact ggagcctgag gagttcgctg cctacctgga
     1381 gccattcaag gagaagggcg aggttagacg gcctaccctc tcctggcctc gcgagatccc
     1441 tctcgttaag ggaggcaagc ccgacgtcgt ccagattgtc cgcaactaca acgcctacct
     1501 tcgggccagc gacgatctgc ctaagatgtt catcgagtcc gaccctgggt tcttttccaa
     1561 cgctattgtc gagggagcta agaagttccc taacaccgag ttcgtgaagg tgaagggcct
     1621 ccacttcagc caggaggacg ctccagatga aatgggtaag tacatcaaga gcttcgtgga
     1681 gcgcgtgctg aagaacgagc agtaattcta ggcgatcgct cgagcccggg aattcgttta
     1741 aacctagagc ggccgctggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg
     1801 gttttttgtg tgaggatcta aatgagtctt cggacctcgc gggggccgct taagcggtgg
     1861 ttagggtttg tctgacgcgg ggggaggggg aaggaacgaa acactctcat tcggaggcgg
     1921 ctcggggttt ggtcttggtg gccacgggca cgcagaagag cgccgcgatc ctcttaagca
     1981 cccccccgcc ctccgtggag gcgggggttt ggtcggcggg tggtaactgg cgggccgctg
     2041 actcgggcgg gtcgcgcgcc ccagagtgtg accttttcgg tctgctcgca gacccccggg
     2101 cggcgccgcc gcggcggcga cgggctcgct gggtcctagg ctccatgggg accgtatacg
     2161 tggacaggct ctggagcatc cgcacgactg cggtgatatt accggagacc ttctgcggga
     2221 cgagccgggt cacgcggctg acgcggagcg tccgttgggc gacaaacacc aggacggggc
     2281 acaggtacac tatcttgtca cccggaggcg cgagggactg caggagcttc agggagtggc
     2341 gcagctgctt catccccgtg gcccgttgct cgcgtttgct ggcggtgtcc ccggaagaaa
     2401 tatatttgca tgtctttagt tctatgatga cacaaacccc gcccagcgtc ttgtcattgg
     2461 cgaattcgaa cacgcagatg cagtcggggc ggcgcggtcc caggtccact tcgcatatta
     2521 aggtgacgcg tgtggcctcg aacaccgagc gaccctgcag cgacccgctt aaaagcttgg
     2581 cattccggta ctgttggtaa agccaccatg gccgatgcta agaacattaa gaagggccct
     2641 gctcccttct accctctgga ggatggcacc gctggcgagc agctgcacaa ggccatgaag
     2701 aggtatgccc tggtgcctgg caccattgcc ttcaccgatg cccacattga ggtggacatc
     2761 acctatgccg agtacttcga gatgtctgtg cgcctggccg aggccatgaa gaggtacggc
     2821 ctgaacacca accaccgcat cgtggtgtgc tctgagaact ctctgcagtt cttcatgcca
     2881 gtgctgggcg ccctgttcat cggagtggcc gtggcccctg ctaacgacat ttacaacgag
     2941 cgcgagctgc tgaacagcat gggcatttct cagcctaccg tggtgttcgt gtctaagaag
     3001 ggcctgcaga agatcctgaa cgtgcagaag aagctgccta tcatccagaa gatcatcatc
     3061 atggactcta agaccgacta ccagggcttc cagagcatgt acacattcgt gacatctcat
     3121 ctgcctcctg gcttcaacga gtacgacttc gtgccagagt ctttcgacag ggacaaaacc
     3181 attgccctga tcatgaacag ctctgggtct accggcctgc ctaagggcgt ggccctgcct
     3241 catcgcaccg cctgtgtgcg cttctctcac gcccgcgacc ctattttcgg caaccagatc
     3301 atccccgaca ccgctattct gagcgtggtg ccattccacc acggcttcgg catgttcacc
     3361 accctgggct acctgatttg cggctttcgg gtggtgctga tgtaccgctt cgaggaggag
     3421 ctgttcctgc gcagcctgca agactacaaa attcagtctg ccctgctggt gccaaccctg
     3481 ttcagcttct tcgctaagag caccctgatc gacaagtacg acctgtctaa cctgcacgag
     3541 attgcctctg gcggcgcccc actgtctaag gaggtgggcg aagccgtggc caagcgcttt
     3601 catctgccag gcatccgcca gggctacggc ctgaccgaga caaccagcgc cattctgatt
     3661 accccagagg gcgacgacaa gcctggcgcc gtgggcaagg tggtgccatt cttcgaggcc
     3721 aaggtggtgg acctggacac cggcaagacc ctgggagtga accagcgcgg cgagctgtgt
     3781 gtgcgcggcc ctatgattat gtccggctac gtgaataacc ctgaggccac aaacgccctg
     3841 atcgacaagg acggctggct gcactctggc gacattgcct actgggacga ggacgagcac
     3901 ttcttcatcg tggaccgcct gaagtctctg atcaagtaca agggctacca ggtggcccca
     3961 gccgagctgg agtctatcct gctgcagcac cctaacattt tcgacgccgg agtggccggc
     4021 ctgcccgacg acgatgccgg cgagctgcct gccgccgtcg tcgtgctgga acacggcaag
     4081 accatgaccg agaaggagat cgtggactat gtggccagcc aggtgacaac cgccaagaag
     4141 ctgcgcggcg gagtggtgtt cgtggacgag gtgcccaagg gcctgaccgg caagctggac
     4201 gcccgcaaga tccgcgagat cctgatcaag gctaagaaag gcggcaagat cgccgtgtaa
     4261 taattctaga gtcggggcgg ccggccgctt cgagcagaca tgataagata cattgatgag
     4321 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     4381 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     4441 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     4501 ctctacaaat gtggtaaaat cgataaggat ccaggtggca cttttcgggg aaatgtgcgc
     4561 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4621 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     4681 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     4741 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     4801 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     4861 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     4921 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     4981 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5041 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5101 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5161 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     5221 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5281 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5341 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5401 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5461 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5521 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5581 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5641 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5701 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5761 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5821 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5881 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5941 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6001 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6061 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6121 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6181 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6241 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6301 tttttacggt tcctggcctt ttgctggcct tttgctcaca tggctcgac
//
