LOCUS       sequence-9387-ex        6925 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6925)
  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 Oct 31. ():.
  PUBMED    18978028
REFERENCE   2  (bases 1 to 6925)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "RNA. 2008 
            Oct 31. ():."
COMMENT     Sequence Label: psiCHECK CHAC1 3'UTR
FEATURES             Location/Qualifiers
     source          1..6925
                     /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    2340..2388
                     /note="synthetic polyadenylation signal"
     promoter        2397..3148
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      3178..3187
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3184..4836
                     /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             4143..4151
                     /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    4880..5001
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4891..4910)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4945..4964
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5134..5238
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5239..5307,5308..6099)
                     /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(5457..5476)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6270..6858
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6759..6778
                     /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 cgctcgagca ctccagtgca caagacagac ttgcgaccgc
     1681 ttgagcccac tgagcagata tggtgggtgg ctggaggctt ctctttctca gtccctgcct
     1741 gtctgccagc ctgcagctct cctgcttgac actgacttac tacttgaaac tttatttatt
     1801 gcaccatgtt ggtgtggtgg gcaggtggag ggcctgccct ggacacaggg gccctgctga
     1861 gcagtggccc catcctggaa cttgaccaga ttccccccag tgctgctgct aaccccacac
     1921 cacccaggcc tccacctccc cagggagtct ccaagagcct cgatcctctg ctcactcagc
     1981 ccagccatcc atagccctgg gaattccacc tgccaaggat cccagcaggc tggatgaggg
     2041 atagtagggc atgaggagaa ggagccctgt aaggactgag gccccggcca gcccttctcc
     2101 tccaccagtt ccccagagca gagctggagc tgatgcctgg acacagctgc tgagcctggc
     2161 ctgggcctct tacccacttg gttgttttct tgtccctctg tctgtctgtc tatctacttg
     2221 tctgtctggg ccactcctgc ctgtgtgttg gtctattcct gggaagctca tcactacagg
     2281 ccctggcaac cttcccagtc tgtcccatac tgttacccat aaaagcggcc gctggccgca
     2341 ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgag gatctaaatg
     2401 agtcttcgga cctcgcgggg gccgcttaag cggtggttag ggtttgtctg acgcgggggg
     2461 agggggaagg aacgaaacac tctcattcgg aggcggctcg gggtttggtc ttggtggcca
     2521 cgggcacgca gaagagcgcc gcgatcctct taagcacccc cccgccctcc gtggaggcgg
     2581 gggtttggtc ggcgggtggt aactggcggg ccgctgactc gggcgggtcg cgcgccccag
     2641 agtgtgacct tttcggtctg ctcgcagacc cccgggcggc gccgccgcgg cggcgacggg
     2701 ctcgctgggt cctaggctcc atggggaccg tatacgtgga caggctctgg agcatccgca
     2761 cgactgcggt gatattaccg gagaccttct gcgggacgag ccgggtcacg cggctgacgc
     2821 ggagcgtccg ttgggcgaca aacaccagga cggggcacag gtacactatc ttgtcacccg
     2881 gaggcgcgag ggactgcagg agcttcaggg agtggcgcag ctgcttcatc cccgtggccc
     2941 gttgctcgcg tttgctggcg gtgtccccgg aagaaatata tttgcatgtc tttagttcta
     3001 tgatgacaca aaccccgccc agcgtcttgt cattggcgaa ttcgaacacg cagatgcagt
     3061 cggggcggcg cggtcccagg tccacttcgc atattaaggt gacgcgtgtg gcctcgaaca
     3121 ccgagcgacc ctgcagcgac ccgcttaaaa gcttggcatt ccggtactgt tggtaaagcc
     3181 accatggccg atgctaagaa cattaagaag ggccctgctc ccttctaccc tctggaggat
     3241 ggcaccgctg gcgagcagct gcacaaggcc atgaagaggt atgccctggt gcctggcacc
     3301 attgccttca ccgatgccca cattgaggtg gacatcacct atgccgagta cttcgagatg
     3361 tctgtgcgcc tggccgaggc catgaagagg tacggcctga acaccaacca ccgcatcgtg
     3421 gtgtgctctg agaactctct gcagttcttc atgccagtgc tgggcgccct gttcatcgga
     3481 gtggccgtgg cccctgctaa cgacatttac aacgagcgcg agctgctgaa cagcatgggc
     3541 atttctcagc ctaccgtggt gttcgtgtct aagaagggcc tgcagaagat cctgaacgtg
     3601 cagaagaagc tgcctatcat ccagaagatc atcatcatgg actctaagac cgactaccag
     3661 ggcttccaga gcatgtacac attcgtgaca tctcatctgc ctcctggctt caacgagtac
     3721 gacttcgtgc cagagtcttt cgacagggac aaaaccattg ccctgatcat gaacagctct
     3781 gggtctaccg gcctgcctaa gggcgtggcc ctgcctcatc gcaccgcctg tgtgcgcttc
     3841 tctcacgccc gcgaccctat tttcggcaac cagatcatcc ccgacaccgc tattctgagc
     3901 gtggtgccat tccaccacgg cttcggcatg ttcaccaccc tgggctacct gatttgcggc
     3961 tttcgggtgg tgctgatgta ccgcttcgag gaggagctgt tcctgcgcag cctgcaagac
     4021 tacaaaattc agtctgccct gctggtgcca accctgttca gcttcttcgc taagagcacc
     4081 ctgatcgaca agtacgacct gtctaacctg cacgagattg cctctggcgg cgccccactg
     4141 tctaaggagg tgggcgaagc cgtggccaag cgctttcatc tgccaggcat ccgccagggc
     4201 tacggcctga ccgagacaac cagcgccatt ctgattaccc cagagggcga cgacaagcct
     4261 ggcgccgtgg gcaaggtggt gccattcttc gaggccaagg tggtggacct ggacaccggc
     4321 aagaccctgg gagtgaacca gcgcggcgag ctgtgtgtgc gcggccctat gattatgtcc
     4381 ggctacgtga ataaccctga ggccacaaac gccctgatcg acaaggacgg ctggctgcac
     4441 tctggcgaca ttgcctactg ggacgaggac gagcacttct tcatcgtgga ccgcctgaag
     4501 tctctgatca agtacaaggg ctaccaggtg gccccagccg agctggagtc tatcctgctg
     4561 cagcacccta acattttcga cgccggagtg gccggcctgc ccgacgacga tgccggcgag
     4621 ctgcctgccg ccgtcgtcgt gctggaacac ggcaagacca tgaccgagaa ggagatcgtg
     4681 gactatgtgg ccagccaggt gacaaccgcc aagaagctgc gcggcggagt ggtgttcgtg
     4741 gacgaggtgc ccaagggcct gaccggcaag ctggacgccc gcaagatccg cgagatcctg
     4801 atcaaggcta agaaaggcgg caagatcgcc gtgtaataat tctagagtcg gggcggccgg
     4861 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     4921 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     4981 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     5041 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat
     5101 aaggatccag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5161 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5221 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5281 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5341 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5401 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5461 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5521 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5581 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5641 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5701 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5761 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5821 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5881 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5941 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6001 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6061 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6121 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6181 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6241 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6301 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6361 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6421 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6481 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6541 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6601 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6661 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6721 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6781 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6841 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6901 tggccttttg ctcacatggc tcgac
//
