LOCUS       sequence-9386-ex        6432 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 6432)
  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 6432)
  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 SIRT4 3'UTR
FEATURES             Location/Qualifiers
     source          1..6432
                     /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    1847..1895
                     /note="synthetic polyadenylation signal"
     promoter        1904..2655
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      2685..2694
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2691..4343
                     /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             3650..3658
                     /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    4387..4508
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4398..4417)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4452..4471
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4641..4745
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4746..4814,4815..5606)
                     /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(4964..4983)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5777..6365
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6266..6285
                     /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 cgctcgaggc tgcctttgat agacccatgc tgaccacagc
     1681 ctgatattcc agaacctgga acagggactt tcacttgaat cttgctgcta aatgtaaatg
     1741 ccttctcaaa tgacagattc cagttcccat tcaacagagt agggtgcact gacaaagtat
     1801 agaaggttct aggtatctta atgtgtggat agcggccgct ggccgcaata aaatatcttt
     1861 attttcatta catctgtgtg ttggtttttt gtgtgaggat ctaaatgagt cttcggacct
     1921 cgcgggggcc gcttaagcgg tggttagggt ttgtctgacg cggggggagg gggaaggaac
     1981 gaaacactct cattcggagg cggctcgggg tttggtcttg gtggccacgg gcacgcagaa
     2041 gagcgccgcg atcctcttaa gcaccccccc gccctccgtg gaggcggggg tttggtcggc
     2101 gggtggtaac tggcgggccg ctgactcggg cgggtcgcgc gccccagagt gtgacctttt
     2161 cggtctgctc gcagaccccc gggcggcgcc gccgcggcgg cgacgggctc gctgggtcct
     2221 aggctccatg gggaccgtat acgtggacag gctctggagc atccgcacga ctgcggtgat
     2281 attaccggag accttctgcg ggacgagccg ggtcacgcgg ctgacgcgga gcgtccgttg
     2341 ggcgacaaac accaggacgg ggcacaggta cactatcttg tcacccggag gcgcgaggga
     2401 ctgcaggagc ttcagggagt ggcgcagctg cttcatcccc gtggcccgtt gctcgcgttt
     2461 gctggcggtg tccccggaag aaatatattt gcatgtcttt agttctatga tgacacaaac
     2521 cccgcccagc gtcttgtcat tggcgaattc gaacacgcag atgcagtcgg ggcggcgcgg
     2581 tcccaggtcc acttcgcata ttaaggtgac gcgtgtggcc tcgaacaccg agcgaccctg
     2641 cagcgacccg cttaaaagct tggcattccg gtactgttgg taaagccacc atggccgatg
     2701 ctaagaacat taagaagggc cctgctccct tctaccctct ggaggatggc accgctggcg
     2761 agcagctgca caaggccatg aagaggtatg ccctggtgcc tggcaccatt gccttcaccg
     2821 atgcccacat tgaggtggac atcacctatg ccgagtactt cgagatgtct gtgcgcctgg
     2881 ccgaggccat gaagaggtac ggcctgaaca ccaaccaccg catcgtggtg tgctctgaga
     2941 actctctgca gttcttcatg ccagtgctgg gcgccctgtt catcggagtg gccgtggccc
     3001 ctgctaacga catttacaac gagcgcgagc tgctgaacag catgggcatt tctcagccta
     3061 ccgtggtgtt cgtgtctaag aagggcctgc agaagatcct gaacgtgcag aagaagctgc
     3121 ctatcatcca gaagatcatc atcatggact ctaagaccga ctaccagggc ttccagagca
     3181 tgtacacatt cgtgacatct catctgcctc ctggcttcaa cgagtacgac ttcgtgccag
     3241 agtctttcga cagggacaaa accattgccc tgatcatgaa cagctctggg tctaccggcc
     3301 tgcctaaggg cgtggccctg cctcatcgca ccgcctgtgt gcgcttctct cacgcccgcg
     3361 accctatttt cggcaaccag atcatccccg acaccgctat tctgagcgtg gtgccattcc
     3421 accacggctt cggcatgttc accaccctgg gctacctgat ttgcggcttt cgggtggtgc
     3481 tgatgtaccg cttcgaggag gagctgttcc tgcgcagcct gcaagactac aaaattcagt
     3541 ctgccctgct ggtgccaacc ctgttcagct tcttcgctaa gagcaccctg atcgacaagt
     3601 acgacctgtc taacctgcac gagattgcct ctggcggcgc cccactgtct aaggaggtgg
     3661 gcgaagccgt ggccaagcgc tttcatctgc caggcatccg ccagggctac ggcctgaccg
     3721 agacaaccag cgccattctg attaccccag agggcgacga caagcctggc gccgtgggca
     3781 aggtggtgcc attcttcgag gccaaggtgg tggacctgga caccggcaag accctgggag
     3841 tgaaccagcg cggcgagctg tgtgtgcgcg gccctatgat tatgtccggc tacgtgaata
     3901 accctgaggc cacaaacgcc ctgatcgaca aggacggctg gctgcactct ggcgacattg
     3961 cctactggga cgaggacgag cacttcttca tcgtggaccg cctgaagtct ctgatcaagt
     4021 acaagggcta ccaggtggcc ccagccgagc tggagtctat cctgctgcag caccctaaca
     4081 ttttcgacgc cggagtggcc ggcctgcccg acgacgatgc cggcgagctg cctgccgccg
     4141 tcgtcgtgct ggaacacggc aagaccatga ccgagaagga gatcgtggac tatgtggcca
     4201 gccaggtgac aaccgccaag aagctgcgcg gcggagtggt gttcgtggac gaggtgccca
     4261 agggcctgac cggcaagctg gacgcccgca agatccgcga gatcctgatc aaggctaaga
     4321 aaggcggcaa gatcgccgtg taataattct agagtcgggg cggccggccg cttcgagcag
     4381 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     4441 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     4501 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     4561 aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatccaggtg
     4621 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4681 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4741 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4801 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4861 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4921 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4981 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5041 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5101 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5161 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5221 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5281 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5341 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5401 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5461 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5521 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5581 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     5641 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5701 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5761 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5821 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5881 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5941 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6001 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6061 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6121 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6181 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6241 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6301 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6361 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6421 acatggctcg ac
//
