LOCUS       sequence-9383-ex        6466 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 6466)
  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 6466)
  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 TTLL11 3'UTR
FEATURES             Location/Qualifiers
     source          1..6466
                     /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    1881..1929
                     /note="synthetic polyadenylation signal"
     promoter        1938..2689
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      2719..2728
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2725..4377
                     /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             3684..3692
                     /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    4421..4542
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4432..4451)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4486..4505
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4675..4779
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4780..4848,4849..5640)
                     /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(4998..5017)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5811..6399
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6300..6319
                     /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 cgctcgagtc aagagggctc tctggcttcg tgtaggatga
     1681 gtgatcgggg tgggatggcc agaggagggg acagtgttgg tggtgctgtg ggcttgggtg
     1741 ttaaacctgg gcccaacttc caagctttct gtttgctccc tgtgtgattt ggggcaagct
     1801 acttcccctc tgttacctgt aaaatgaaat gaagaatacc tatcttaaaa ggttcttgta
     1861 agaatgcggc cgctggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     1921 ttttgtgtga ggatctaaat gagtcttcgg acctcgcggg ggccgcttaa gcggtggtta
     1981 gggtttgtct gacgcggggg gagggggaag gaacgaaaca ctctcattcg gaggcggctc
     2041 ggggtttggt cttggtggcc acgggcacgc agaagagcgc cgcgatcctc ttaagcaccc
     2101 ccccgccctc cgtggaggcg ggggtttggt cggcgggtgg taactggcgg gccgctgact
     2161 cgggcgggtc gcgcgcccca gagtgtgacc ttttcggtct gctcgcagac ccccgggcgg
     2221 cgccgccgcg gcggcgacgg gctcgctggg tcctaggctc catggggacc gtatacgtgg
     2281 acaggctctg gagcatccgc acgactgcgg tgatattacc ggagaccttc tgcgggacga
     2341 gccgggtcac gcggctgacg cggagcgtcc gttgggcgac aaacaccagg acggggcaca
     2401 ggtacactat cttgtcaccc ggaggcgcga gggactgcag gagcttcagg gagtggcgca
     2461 gctgcttcat ccccgtggcc cgttgctcgc gtttgctggc ggtgtccccg gaagaaatat
     2521 atttgcatgt ctttagttct atgatgacac aaaccccgcc cagcgtcttg tcattggcga
     2581 attcgaacac gcagatgcag tcggggcggc gcggtcccag gtccacttcg catattaagg
     2641 tgacgcgtgt ggcctcgaac accgagcgac cctgcagcga cccgcttaaa agcttggcat
     2701 tccggtactg ttggtaaagc caccatggcc gatgctaaga acattaagaa gggccctgct
     2761 cccttctacc ctctggagga tggcaccgct ggcgagcagc tgcacaaggc catgaagagg
     2821 tatgccctgg tgcctggcac cattgccttc accgatgccc acattgaggt ggacatcacc
     2881 tatgccgagt acttcgagat gtctgtgcgc ctggccgagg ccatgaagag gtacggcctg
     2941 aacaccaacc accgcatcgt ggtgtgctct gagaactctc tgcagttctt catgccagtg
     3001 ctgggcgccc tgttcatcgg agtggccgtg gcccctgcta acgacattta caacgagcgc
     3061 gagctgctga acagcatggg catttctcag cctaccgtgg tgttcgtgtc taagaagggc
     3121 ctgcagaaga tcctgaacgt gcagaagaag ctgcctatca tccagaagat catcatcatg
     3181 gactctaaga ccgactacca gggcttccag agcatgtaca cattcgtgac atctcatctg
     3241 cctcctggct tcaacgagta cgacttcgtg ccagagtctt tcgacaggga caaaaccatt
     3301 gccctgatca tgaacagctc tgggtctacc ggcctgccta agggcgtggc cctgcctcat
     3361 cgcaccgcct gtgtgcgctt ctctcacgcc cgcgacccta ttttcggcaa ccagatcatc
     3421 cccgacaccg ctattctgag cgtggtgcca ttccaccacg gcttcggcat gttcaccacc
     3481 ctgggctacc tgatttgcgg ctttcgggtg gtgctgatgt accgcttcga ggaggagctg
     3541 ttcctgcgca gcctgcaaga ctacaaaatt cagtctgccc tgctggtgcc aaccctgttc
     3601 agcttcttcg ctaagagcac cctgatcgac aagtacgacc tgtctaacct gcacgagatt
     3661 gcctctggcg gcgccccact gtctaaggag gtgggcgaag ccgtggccaa gcgctttcat
     3721 ctgccaggca tccgccaggg ctacggcctg accgagacaa ccagcgccat tctgattacc
     3781 ccagagggcg acgacaagcc tggcgccgtg ggcaaggtgg tgccattctt cgaggccaag
     3841 gtggtggacc tggacaccgg caagaccctg ggagtgaacc agcgcggcga gctgtgtgtg
     3901 cgcggcccta tgattatgtc cggctacgtg aataaccctg aggccacaaa cgccctgatc
     3961 gacaaggacg gctggctgca ctctggcgac attgcctact gggacgagga cgagcacttc
     4021 ttcatcgtgg accgcctgaa gtctctgatc aagtacaagg gctaccaggt ggccccagcc
     4081 gagctggagt ctatcctgct gcagcaccct aacattttcg acgccggagt ggccggcctg
     4141 cccgacgacg atgccggcga gctgcctgcc gccgtcgtcg tgctggaaca cggcaagacc
     4201 atgaccgaga aggagatcgt ggactatgtg gccagccagg tgacaaccgc caagaagctg
     4261 cgcggcggag tggtgttcgt ggacgaggtg cccaagggcc tgaccggcaa gctggacgcc
     4321 cgcaagatcc gcgagatcct gatcaaggct aagaaaggcg gcaagatcgc cgtgtaataa
     4381 ttctagagtc ggggcggccg gccgcttcga gcagacatga taagatacat tgatgagttt
     4441 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     4501 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     4561 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
     4621 tacaaatgtg gtaaaatcga taaggatcca ggtggcactt ttcggggaaa tgtgcgcgga
     4681 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4741 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4801 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4861 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4921 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4981 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     5041 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5101 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5161 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5221 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5281 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5341 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5401 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5461 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5521 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5581 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5641 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5701 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5761 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5821 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5881 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5941 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6001 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6061 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6121 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6181 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6241 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6301 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6361 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6421 ttacggttcc tggccttttg ctggcctttt gctcacatgg ctcgac
//
