LOCUS       sequence-9384-ex        6656 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6656)
  AUTHORS   Landthaler M, Gaidatzis D, Rothballer A, Chen PY, Soll SJ, Dinic L, 
            Ojo T, Hafner M, Zavolan M, Tuschl T
  TITLE     Molecular characterization of human Argonaute-containing 
            ribonucleoprotein complexes and their bound target mRNAs.
  JOURNAL   RNA. 2008 Dec;14(12):2580-96. doi: 10.1261/rna.1351608. Epub 2008 
            Oct 31.
  PUBMED    18978028
REFERENCE   2  (bases 1 to 6656)
  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: 
            "10.1261/rna.1351608"; journalName: "RNA"; date: "2008-12"; volume: 
            "14"; issue: "12"; pages: "2580-96"
COMMENT     Sequence Label: psiCHECK RHEBL1 3'UTR
FEATURES             Location/Qualifiers
     source          1..6656
                     /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"
     promoter        666..684
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      688..697
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             694..1629
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2071..2119
                     /note="synthetic polyadenylation signal"
     promoter        2128..2879
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      2909..2918
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2915..4567
                     /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             3874..3882
                     /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    4611..4732
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4622..4641)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4676..4695
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4865..4969
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4970..5038,5039..5830)
                     /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(5188..5207)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6001..6589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6490..6509
                     /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 taggctagcc accatggctt ccaaggtgta cgaccccgag
      721 caacgcaaac gcatgatcac tgggcctcag tggtgggctc gctgcaagca aatgaacgtg
      781 ctggactcct tcatcaacta ctatgattcc gagaagcacg ccgagaacgc cgtgattttt
      841 ctgcatggta acgctgcctc cagctacctg tggaggcacg tcgtgcctca catcgagccc
      901 gtggctagat gcatcatccc tgatctgatc ggaatgggta agtccggcaa gagcgggaat
      961 ggctcatatc gcctcctgga tcactacaag tacctcaccg cttggttcga gctgctgaac
     1021 cttccaaaga aaatcatctt tgtgggccac gactgggggg cttgtctggc ctttcactac
     1081 tcctacgagc accaagacaa gatcaaggcc atcgtccatg ctgagagtgt cgtggacgtg
     1141 atcgagtcct gggacgagtg gcctgacatc gaggaggata tcgccctgat caagagcgaa
     1201 gagggcgaga aaatggtgct tgagaataac ttcttcgtcg agaccatgct cccaagcaag
     1261 atcatgcgga aactggagcc tgaggagttc gctgcctacc tggagccatt caaggagaag
     1321 ggcgaggtta gacggcctac cctctcctgg cctcgcgaga tccctctcgt taagggaggc
     1381 aagcccgacg tcgtccagat tgtccgcaac tacaacgcct accttcgggc cagcgacgat
     1441 ctgcctaaga tgttcatcga gtccgaccct gggttctttt ccaacgctat tgtcgaggga
     1501 gctaagaagt tccctaacac cgagttcgtg aaggtgaagg gcctccactt cagccaggag
     1561 gacgctccag atgaaatggg taagtacatc aagagcttcg tggagcgcgt gctgaagaac
     1621 gagcagtaat tctaggcgat cgctcgaggt gtggggtaac tgccttgctt ctgcccccgg
     1681 cacttgccat gttccagtgg ggggcagatc ctcaggactt cacgggtatg gttgccagct
     1741 gtgttcctgg cccctggaca cacagtgtgg catcctcatg tttgcacact ttccccaggc
     1801 tccagtggcc tggatgtcaa tgtttacaaa ggggcaagga cctctcatgg acactggcct
     1861 ctagccctct gtttttgttt gatgaattct gttataacct atggggtcag gatatgagtc
     1921 ctgggcatta tttatccagg acccatcctc ttgggtgggt tttgggtgtt ggctgggtaa
     1981 ggggagccgg ggacttctga aatagagctg gctccctggg gtgacaatgt atatatgcaa
     2041 ataaattgag aaatcgcggc cgctggccgc aataaaatat ctttattttc attacatctg
     2101 tgtgttggtt ttttgtgtga ggatctaaat gagtcttcgg acctcgcggg ggccgcttaa
     2161 gcggtggtta gggtttgtct gacgcggggg gagggggaag gaacgaaaca ctctcattcg
     2221 gaggcggctc ggggtttggt cttggtggcc acgggcacgc agaagagcgc cgcgatcctc
     2281 ttaagcaccc ccccgccctc cgtggaggcg ggggtttggt cggcgggtgg taactggcgg
     2341 gccgctgact cgggcgggtc gcgcgcccca gagtgtgacc ttttcggtct gctcgcagac
     2401 ccccgggcgg cgccgccgcg gcggcgacgg gctcgctggg tcctaggctc catggggacc
     2461 gtatacgtgg acaggctctg gagcatccgc acgactgcgg tgatattacc ggagaccttc
     2521 tgcgggacga gccgggtcac gcggctgacg cggagcgtcc gttgggcgac aaacaccagg
     2581 acggggcaca ggtacactat cttgtcaccc ggaggcgcga gggactgcag gagcttcagg
     2641 gagtggcgca gctgcttcat ccccgtggcc cgttgctcgc gtttgctggc ggtgtccccg
     2701 gaagaaatat atttgcatgt ctttagttct atgatgacac aaaccccgcc cagcgtcttg
     2761 tcattggcga attcgaacac gcagatgcag tcggggcggc gcggtcccag gtccacttcg
     2821 catattaagg tgacgcgtgt ggcctcgaac accgagcgac cctgcagcga cccgcttaaa
     2881 agcttggcat tccggtactg ttggtaaagc caccatggcc gatgctaaga acattaagaa
     2941 gggccctgct cccttctacc ctctggagga tggcaccgct ggcgagcagc tgcacaaggc
     3001 catgaagagg tatgccctgg tgcctggcac cattgccttc accgatgccc acattgaggt
     3061 ggacatcacc tatgccgagt acttcgagat gtctgtgcgc ctggccgagg ccatgaagag
     3121 gtacggcctg aacaccaacc accgcatcgt ggtgtgctct gagaactctc tgcagttctt
     3181 catgccagtg ctgggcgccc tgttcatcgg agtggccgtg gcccctgcta acgacattta
     3241 caacgagcgc gagctgctga acagcatggg catttctcag cctaccgtgg tgttcgtgtc
     3301 taagaagggc ctgcagaaga tcctgaacgt gcagaagaag ctgcctatca tccagaagat
     3361 catcatcatg gactctaaga ccgactacca gggcttccag agcatgtaca cattcgtgac
     3421 atctcatctg cctcctggct tcaacgagta cgacttcgtg ccagagtctt tcgacaggga
     3481 caaaaccatt gccctgatca tgaacagctc tgggtctacc ggcctgccta agggcgtggc
     3541 cctgcctcat cgcaccgcct gtgtgcgctt ctctcacgcc cgcgacccta ttttcggcaa
     3601 ccagatcatc cccgacaccg ctattctgag cgtggtgcca ttccaccacg gcttcggcat
     3661 gttcaccacc ctgggctacc tgatttgcgg ctttcgggtg gtgctgatgt accgcttcga
     3721 ggaggagctg ttcctgcgca gcctgcaaga ctacaaaatt cagtctgccc tgctggtgcc
     3781 aaccctgttc agcttcttcg ctaagagcac cctgatcgac aagtacgacc tgtctaacct
     3841 gcacgagatt gcctctggcg gcgccccact gtctaaggag gtgggcgaag ccgtggccaa
     3901 gcgctttcat ctgccaggca tccgccaggg ctacggcctg accgagacaa ccagcgccat
     3961 tctgattacc ccagagggcg acgacaagcc tggcgccgtg ggcaaggtgg tgccattctt
     4021 cgaggccaag gtggtggacc tggacaccgg caagaccctg ggagtgaacc agcgcggcga
     4081 gctgtgtgtg cgcggcccta tgattatgtc cggctacgtg aataaccctg aggccacaaa
     4141 cgccctgatc gacaaggacg gctggctgca ctctggcgac attgcctact gggacgagga
     4201 cgagcacttc ttcatcgtgg accgcctgaa gtctctgatc aagtacaagg gctaccaggt
     4261 ggccccagcc gagctggagt ctatcctgct gcagcaccct aacattttcg acgccggagt
     4321 ggccggcctg cccgacgacg atgccggcga gctgcctgcc gccgtcgtcg tgctggaaca
     4381 cggcaagacc atgaccgaga aggagatcgt ggactatgtg gccagccagg tgacaaccgc
     4441 caagaagctg cgcggcggag tggtgttcgt ggacgaggtg cccaagggcc tgaccggcaa
     4501 gctggacgcc cgcaagatcc gcgagatcct gatcaaggct aagaaaggcg gcaagatcgc
     4561 cgtgtaataa ttctagagtc ggggcggccg gccgcttcga gcagacatga taagatacat
     4621 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     4681 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     4741 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     4801 gtaaaacctc tacaaatgtg gtaaaatcga taaggatcca ggtggcactt ttcggggaaa
     4861 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4921 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4981 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     5041 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     5101 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     5161 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     5221 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     5281 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     5341 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     5401 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5461 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     5521 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     5581 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5641 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5701 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5761 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5821 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     5881 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5941 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6001 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6061 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     6181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgg ctcgac
//
