LOCUS       sequence-9380-ex        7543 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 7543)
  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 7543)
  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 ENPP5 3'UTR
FEATURES             Location/Qualifiers
     source          1..7543
                     /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    2958..3006
                     /note="synthetic polyadenylation signal"
     promoter        3015..3766
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      3796..3805
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3802..5454
                     /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             4761..4769
                     /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    5498..5619
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5509..5528)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5563..5582
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5752..5856
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5857..5925,5926..6717)
                     /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(6075..6094)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6888..7476
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7377..7396
                     /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 cgctcgagag atatgcatgc tgaaatagct caaccattat
     1681 tacaagccta atgttacttt gaagtggatt tgcatattga agtggagatt ccataattat
     1741 gtcagtgttt aaaggtttca aattctggga aaccagttcc aaacatttgc agaaaccatt
     1801 aagcagttac atatttaggt atacacacac acacacacac acatacacac acacggacca
     1861 aaatacttac acctgcaaag gaataaagat gtgagagtat gtctccattg ttcactgtag
     1921 catagggata gataagatcc tgctttattt ggacttggcg cagataatgt atatatttag
     1981 caactttgca ctatgtaaag taccttatgt attgcacttt aaatttctct cctgatgggt
     2041 actttaattt gaaatgcact ttatgcacag ttatgtctta taacttgatt gaaaatgaca
     2101 actttttgca cccatgtcac agaatacttg ttacgcattg ttcaaactga aggaaatttc
     2161 taataatccc gaataatgaa cgtagaaatc tatctccata aattgagaga agaagaaggt
     2221 gataagtgtt gaaaattaaa tgtgataacc tttgaacctt gaattttgga gatgtattcc
     2281 caacagcaga atgcaactgt gggcatttct tgtcttattt ctttccagag aacgtggttt
     2341 tcatttattt ttccctcaaa agagagtcaa atactgacag attcgttcta aatatattgt
     2401 ttctgtcata aaattattgt gatttcctga tgagtcatat tactgtgatt ttcataataa
     2461 tgaagacacc atgaatatac tttttttcta tatagttcag caatggcctg aatagaagca
     2521 accaggcacc atctcagcaa tgttttctct tgtttgtaat tatttgctcc tttgaaaatt
     2581 aaatcactat taattacatt aaaaatcaaa ttggataaaa caatgttttc tttctggtag
     2641 cgcataataa cagagcacaa gcatctttta gatttgagca tttgaagatt caaagttgcg
     2701 aaagagcaca aaccatatta ggtaaaatat tggccactcg atccttgaaa agaactgtgt
     2761 ggagcctgga aaaaaaaatt aggaccacat gtgagatgtt tacaagacac ccagaacagt
     2821 ggtaaagtgt gcatactaga aaaagcagca aaataactct ttgtggtaac aggtatcaaa
     2881 acacgggagc cgagatgata tagctctgtt tcaagaaata tgtgaatacc cacctaccag
     2941 gtgcggccgc tggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt
     3001 tgtgtgagga tctaaatgag tcttcggacc tcgcgggggc cgcttaagcg gtggttaggg
     3061 tttgtctgac gcggggggag ggggaaggaa cgaaacactc tcattcggag gcggctcggg
     3121 gtttggtctt ggtggccacg ggcacgcaga agagcgccgc gatcctctta agcacccccc
     3181 cgccctccgt ggaggcgggg gtttggtcgg cgggtggtaa ctggcgggcc gctgactcgg
     3241 gcgggtcgcg cgccccagag tgtgaccttt tcggtctgct cgcagacccc cgggcggcgc
     3301 cgccgcggcg gcgacgggct cgctgggtcc taggctccat ggggaccgta tacgtggaca
     3361 ggctctggag catccgcacg actgcggtga tattaccgga gaccttctgc gggacgagcc
     3421 gggtcacgcg gctgacgcgg agcgtccgtt gggcgacaaa caccaggacg gggcacaggt
     3481 acactatctt gtcacccgga ggcgcgaggg actgcaggag cttcagggag tggcgcagct
     3541 gcttcatccc cgtggcccgt tgctcgcgtt tgctggcggt gtccccggaa gaaatatatt
     3601 tgcatgtctt tagttctatg atgacacaaa ccccgcccag cgtcttgtca ttggcgaatt
     3661 cgaacacgca gatgcagtcg gggcggcgcg gtcccaggtc cacttcgcat attaaggtga
     3721 cgcgtgtggc ctcgaacacc gagcgaccct gcagcgaccc gcttaaaagc ttggcattcc
     3781 ggtactgttg gtaaagccac catggccgat gctaagaaca ttaagaaggg ccctgctccc
     3841 ttctaccctc tggaggatgg caccgctggc gagcagctgc acaaggccat gaagaggtat
     3901 gccctggtgc ctggcaccat tgccttcacc gatgcccaca ttgaggtgga catcacctat
     3961 gccgagtact tcgagatgtc tgtgcgcctg gccgaggcca tgaagaggta cggcctgaac
     4021 accaaccacc gcatcgtggt gtgctctgag aactctctgc agttcttcat gccagtgctg
     4081 ggcgccctgt tcatcggagt ggccgtggcc cctgctaacg acatttacaa cgagcgcgag
     4141 ctgctgaaca gcatgggcat ttctcagcct accgtggtgt tcgtgtctaa gaagggcctg
     4201 cagaagatcc tgaacgtgca gaagaagctg cctatcatcc agaagatcat catcatggac
     4261 tctaagaccg actaccaggg cttccagagc atgtacacat tcgtgacatc tcatctgcct
     4321 cctggcttca acgagtacga cttcgtgcca gagtctttcg acagggacaa aaccattgcc
     4381 ctgatcatga acagctctgg gtctaccggc ctgcctaagg gcgtggccct gcctcatcgc
     4441 accgcctgtg tgcgcttctc tcacgcccgc gaccctattt tcggcaacca gatcatcccc
     4501 gacaccgcta ttctgagcgt ggtgccattc caccacggct tcggcatgtt caccaccctg
     4561 ggctacctga tttgcggctt tcgggtggtg ctgatgtacc gcttcgagga ggagctgttc
     4621 ctgcgcagcc tgcaagacta caaaattcag tctgccctgc tggtgccaac cctgttcagc
     4681 ttcttcgcta agagcaccct gatcgacaag tacgacctgt ctaacctgca cgagattgcc
     4741 tctggcggcg ccccactgtc taaggaggtg ggcgaagccg tggccaagcg ctttcatctg
     4801 ccaggcatcc gccagggcta cggcctgacc gagacaacca gcgccattct gattacccca
     4861 gagggcgacg acaagcctgg cgccgtgggc aaggtggtgc cattcttcga ggccaaggtg
     4921 gtggacctgg acaccggcaa gaccctggga gtgaaccagc gcggcgagct gtgtgtgcgc
     4981 ggccctatga ttatgtccgg ctacgtgaat aaccctgagg ccacaaacgc cctgatcgac
     5041 aaggacggct ggctgcactc tggcgacatt gcctactggg acgaggacga gcacttcttc
     5101 atcgtggacc gcctgaagtc tctgatcaag tacaagggct accaggtggc cccagccgag
     5161 ctggagtcta tcctgctgca gcaccctaac attttcgacg ccggagtggc cggcctgccc
     5221 gacgacgatg ccggcgagct gcctgccgcc gtcgtcgtgc tggaacacgg caagaccatg
     5281 accgagaagg agatcgtgga ctatgtggcc agccaggtga caaccgccaa gaagctgcgc
     5341 ggcggagtgg tgttcgtgga cgaggtgccc aagggcctga ccggcaagct ggacgcccgc
     5401 aagatccgcg agatcctgat caaggctaag aaaggcggca agatcgccgt gtaataattc
     5461 tagagtcggg gcggccggcc gcttcgagca gacatgataa gatacattga tgagtttgga
     5521 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     5581 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     5641 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     5701 aaatgtggta aaatcgataa ggatccaggt ggcacttttc ggggaaatgt gcgcggaacc
     5761 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5821 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5881 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5941 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     6001 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     6061 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     6121 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     6181 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     6241 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     6301 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     6361 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     6421 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6481 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6541 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6601 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6661 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6721 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6781 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6841 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6901 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6961 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     7021 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     7081 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7141 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7201 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7261 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7321 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7381 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7441 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7501 cggttcctgg ccttttgctg gccttttgct cacatggctc gac
//
