LOCUS       sequence-9381-ex        6855 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 6855)
  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 6855)
  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 SCN1B 3'UTR
FEATURES             Location/Qualifiers
     source          1..6855
                     /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    2270..2318
                     /note="synthetic polyadenylation signal"
     promoter        2327..3078
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      3108..3117
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3114..4766
                     /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             4073..4081
                     /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    4810..4931
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4821..4840)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4875..4894
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5064..5168
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5169..5237,5238..6029)
                     /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(5387..5406)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6200..6788
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6689..6708
                     /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 cgctcgagct caaggaagag ccagccgtaa tggggactct
     1681 ccaggcaccg cctgccccca gcgtgggggt ggccactcct gggccccaga aagcctcaga
     1741 gtcctgccga cggagccact ggggtgggag ggggcagggg gcttggctcg cacccccact
     1801 ttcgcctcct ccagctcctg ccccgccggc cgcgcaccgc catgcatgat gggtaaagca
     1861 atactgccgc tgcccccacc ctgcttctgc tgcctgtttg gggagggggg cggtgaggtg
     1921 ggggcagcgg ccccgcaccc ctcctccttg ctgatttgca cacattggcc gcttcagaca
     1981 cgcacttctg gggccagccc ctccccgcct cctccctgcc tggcggcagg ggtcgcgatg
     2041 atgggctgga gcagtttggg gcagggggtt ctgggaccca ctccgactcc ccctccccgg
     2101 catcatttcc cctcccgctt cctccggctg gacctggggt cccccctccc tgtaatgcac
     2161 tcctgccccg gcccaacctc gccctctctc accagccttg aactgtggcc acctagaaag
     2221 gggcccattc agcctcgtct ctttacagaa gtaggcggcc gctggccgca ataaaatatc
     2281 tttattttca ttacatctgt gtgttggttt tttgtgtgag gatctaaatg agtcttcgga
     2341 cctcgcgggg gccgcttaag cggtggttag ggtttgtctg acgcgggggg agggggaagg
     2401 aacgaaacac tctcattcgg aggcggctcg gggtttggtc ttggtggcca cgggcacgca
     2461 gaagagcgcc gcgatcctct taagcacccc cccgccctcc gtggaggcgg gggtttggtc
     2521 ggcgggtggt aactggcggg ccgctgactc gggcgggtcg cgcgccccag agtgtgacct
     2581 tttcggtctg ctcgcagacc cccgggcggc gccgccgcgg cggcgacggg ctcgctgggt
     2641 cctaggctcc atggggaccg tatacgtgga caggctctgg agcatccgca cgactgcggt
     2701 gatattaccg gagaccttct gcgggacgag ccgggtcacg cggctgacgc ggagcgtccg
     2761 ttgggcgaca aacaccagga cggggcacag gtacactatc ttgtcacccg gaggcgcgag
     2821 ggactgcagg agcttcaggg agtggcgcag ctgcttcatc cccgtggccc gttgctcgcg
     2881 tttgctggcg gtgtccccgg aagaaatata tttgcatgtc tttagttcta tgatgacaca
     2941 aaccccgccc agcgtcttgt cattggcgaa ttcgaacacg cagatgcagt cggggcggcg
     3001 cggtcccagg tccacttcgc atattaaggt gacgcgtgtg gcctcgaaca ccgagcgacc
     3061 ctgcagcgac ccgcttaaaa gcttggcatt ccggtactgt tggtaaagcc accatggccg
     3121 atgctaagaa cattaagaag ggccctgctc ccttctaccc tctggaggat ggcaccgctg
     3181 gcgagcagct gcacaaggcc atgaagaggt atgccctggt gcctggcacc attgccttca
     3241 ccgatgccca cattgaggtg gacatcacct atgccgagta cttcgagatg tctgtgcgcc
     3301 tggccgaggc catgaagagg tacggcctga acaccaacca ccgcatcgtg gtgtgctctg
     3361 agaactctct gcagttcttc atgccagtgc tgggcgccct gttcatcgga gtggccgtgg
     3421 cccctgctaa cgacatttac aacgagcgcg agctgctgaa cagcatgggc atttctcagc
     3481 ctaccgtggt gttcgtgtct aagaagggcc tgcagaagat cctgaacgtg cagaagaagc
     3541 tgcctatcat ccagaagatc atcatcatgg actctaagac cgactaccag ggcttccaga
     3601 gcatgtacac attcgtgaca tctcatctgc ctcctggctt caacgagtac gacttcgtgc
     3661 cagagtcttt cgacagggac aaaaccattg ccctgatcat gaacagctct gggtctaccg
     3721 gcctgcctaa gggcgtggcc ctgcctcatc gcaccgcctg tgtgcgcttc tctcacgccc
     3781 gcgaccctat tttcggcaac cagatcatcc ccgacaccgc tattctgagc gtggtgccat
     3841 tccaccacgg cttcggcatg ttcaccaccc tgggctacct gatttgcggc tttcgggtgg
     3901 tgctgatgta ccgcttcgag gaggagctgt tcctgcgcag cctgcaagac tacaaaattc
     3961 agtctgccct gctggtgcca accctgttca gcttcttcgc taagagcacc ctgatcgaca
     4021 agtacgacct gtctaacctg cacgagattg cctctggcgg cgccccactg tctaaggagg
     4081 tgggcgaagc cgtggccaag cgctttcatc tgccaggcat ccgccagggc tacggcctga
     4141 ccgagacaac cagcgccatt ctgattaccc cagagggcga cgacaagcct ggcgccgtgg
     4201 gcaaggtggt gccattcttc gaggccaagg tggtggacct ggacaccggc aagaccctgg
     4261 gagtgaacca gcgcggcgag ctgtgtgtgc gcggccctat gattatgtcc ggctacgtga
     4321 ataaccctga ggccacaaac gccctgatcg acaaggacgg ctggctgcac tctggcgaca
     4381 ttgcctactg ggacgaggac gagcacttct tcatcgtgga ccgcctgaag tctctgatca
     4441 agtacaaggg ctaccaggtg gccccagccg agctggagtc tatcctgctg cagcacccta
     4501 acattttcga cgccggagtg gccggcctgc ccgacgacga tgccggcgag ctgcctgccg
     4561 ccgtcgtcgt gctggaacac ggcaagacca tgaccgagaa ggagatcgtg gactatgtgg
     4621 ccagccaggt gacaaccgcc aagaagctgc gcggcggagt ggtgttcgtg gacgaggtgc
     4681 ccaagggcct gaccggcaag ctggacgccc gcaagatccg cgagatcctg atcaaggcta
     4741 agaaaggcgg caagatcgcc gtgtaataat tctagagtcg gggcggccgg ccgcttcgag
     4801 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     4861 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     4921 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     4981 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatccag
     5041 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     5101 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     5161 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     5221 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     5281 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     5341 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     5401 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     5461 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     5521 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     5581 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     5641 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     5701 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     5761 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     5821 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5881 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5941 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     6001 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     6061 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     6121 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     6181 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     6241 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     6301 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     6361 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6421 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6481 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6541 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6601 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6661 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6721 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6781 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6841 ctcacatggc tcgac
//
