LOCUS       sequence-9391-ex        7218 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 7218)
  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 7218)
  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 SGK 3'UTR
FEATURES             Location/Qualifiers
     source          1..7218
                     /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    2633..2681
                     /note="synthetic polyadenylation signal"
     promoter        2690..3441
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      3471..3480
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3477..5129
                     /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             4436..4444
                     /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    5173..5294
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5184..5203)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5238..5257
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5427..5531
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5532..5600,5601..6392)
                     /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(5750..5769)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6563..7151
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7052..7071
                     /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 cgctcgagga ctctttcctc tgaaccctgt tagggcttgg
     1681 ttttaaagga ttttatgtgt gtttccgaat gttttagtta gccttttggt ggagccgcca
     1741 gctgacagga catcttacaa gagaatttgc acatctctgg aagcttagca atcttattgc
     1801 acactgttcg ctggaagctt tttgaagagc acattctcct cagtgagctc atgaggtttt
     1861 catttttatt cttccttcca acgtggtgct atctctgaaa cgagcgttag agtgccgcct
     1921 tagacggagg caggagtttc gttagaaagc ggacgctgtt ctaaaaaagg tctcctgcag
     1981 atctgtctgg gctgtgatga cgaatattat gaaatgtgcc ttttctgaag agattgtgtt
     2041 agctccaaag cttttcctat cgcagtgttt cagttcttta ttttcccttg tggatatgct
     2101 gtgtgaaccg tcgtgtgagt gtggtatgcc tgatcacaga tggattttgt tataagcatc
     2161 aatgtgacac ttgcaggaca ctacaacgtg ggacattgtt tgtttcttcc atatttggaa
     2221 gataaattta tgtgtagact tttttgtaag atacggttaa taactaaaat ttattgaaat
     2281 ggtcttgcaa tgactcgtat tcagatgctt aaagaaagca ttgctgctac aaatatttct
     2341 atttttagaa agggttttta tggaccaatg ccccagttgt cagtcagagc cgttggtgtt
     2401 tttcattgtt taaaatgtca cctgtaaaat gggcattatt tatgtttttt tttttgcatt
     2461 cctgataatt gtatgtattg tataaagaac gtctgtacat tgggttataa cactagtata
     2521 tttaaactta caggcttatt tgtaatgtaa accaccattt taatgtactg taattaacat
     2581 ggttataata cgtacaatcc ttccctcatc ccatcacgcg gccgctggcc gcaataaaat
     2641 atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaggatctaa atgagtcttc
     2701 ggacctcgcg ggggccgctt aagcggtggt tagggtttgt ctgacgcggg gggaggggga
     2761 aggaacgaaa cactctcatt cggaggcggc tcggggtttg gtcttggtgg ccacgggcac
     2821 gcagaagagc gccgcgatcc tcttaagcac ccccccgccc tccgtggagg cgggggtttg
     2881 gtcggcgggt ggtaactggc gggccgctga ctcgggcggg tcgcgcgccc cagagtgtga
     2941 ccttttcggt ctgctcgcag acccccgggc ggcgccgccg cggcggcgac gggctcgctg
     3001 ggtcctaggc tccatgggga ccgtatacgt ggacaggctc tggagcatcc gcacgactgc
     3061 ggtgatatta ccggagacct tctgcgggac gagccgggtc acgcggctga cgcggagcgt
     3121 ccgttgggcg acaaacacca ggacggggca caggtacact atcttgtcac ccggaggcgc
     3181 gagggactgc aggagcttca gggagtggcg cagctgcttc atccccgtgg cccgttgctc
     3241 gcgtttgctg gcggtgtccc cggaagaaat atatttgcat gtctttagtt ctatgatgac
     3301 acaaaccccg cccagcgtct tgtcattggc gaattcgaac acgcagatgc agtcggggcg
     3361 gcgcggtccc aggtccactt cgcatattaa ggtgacgcgt gtggcctcga acaccgagcg
     3421 accctgcagc gacccgctta aaagcttggc attccggtac tgttggtaaa gccaccatgg
     3481 ccgatgctaa gaacattaag aagggccctg ctcccttcta ccctctggag gatggcaccg
     3541 ctggcgagca gctgcacaag gccatgaaga ggtatgccct ggtgcctggc accattgcct
     3601 tcaccgatgc ccacattgag gtggacatca cctatgccga gtacttcgag atgtctgtgc
     3661 gcctggccga ggccatgaag aggtacggcc tgaacaccaa ccaccgcatc gtggtgtgct
     3721 ctgagaactc tctgcagttc ttcatgccag tgctgggcgc cctgttcatc ggagtggccg
     3781 tggcccctgc taacgacatt tacaacgagc gcgagctgct gaacagcatg ggcatttctc
     3841 agcctaccgt ggtgttcgtg tctaagaagg gcctgcagaa gatcctgaac gtgcagaaga
     3901 agctgcctat catccagaag atcatcatca tggactctaa gaccgactac cagggcttcc
     3961 agagcatgta cacattcgtg acatctcatc tgcctcctgg cttcaacgag tacgacttcg
     4021 tgccagagtc tttcgacagg gacaaaacca ttgccctgat catgaacagc tctgggtcta
     4081 ccggcctgcc taagggcgtg gccctgcctc atcgcaccgc ctgtgtgcgc ttctctcacg
     4141 cccgcgaccc tattttcggc aaccagatca tccccgacac cgctattctg agcgtggtgc
     4201 cattccacca cggcttcggc atgttcacca ccctgggcta cctgatttgc ggctttcggg
     4261 tggtgctgat gtaccgcttc gaggaggagc tgttcctgcg cagcctgcaa gactacaaaa
     4321 ttcagtctgc cctgctggtg ccaaccctgt tcagcttctt cgctaagagc accctgatcg
     4381 acaagtacga cctgtctaac ctgcacgaga ttgcctctgg cggcgcccca ctgtctaagg
     4441 aggtgggcga agccgtggcc aagcgctttc atctgccagg catccgccag ggctacggcc
     4501 tgaccgagac aaccagcgcc attctgatta ccccagaggg cgacgacaag cctggcgccg
     4561 tgggcaaggt ggtgccattc ttcgaggcca aggtggtgga cctggacacc ggcaagaccc
     4621 tgggagtgaa ccagcgcggc gagctgtgtg tgcgcggccc tatgattatg tccggctacg
     4681 tgaataaccc tgaggccaca aacgccctga tcgacaagga cggctggctg cactctggcg
     4741 acattgccta ctgggacgag gacgagcact tcttcatcgt ggaccgcctg aagtctctga
     4801 tcaagtacaa gggctaccag gtggccccag ccgagctgga gtctatcctg ctgcagcacc
     4861 ctaacatttt cgacgccgga gtggccggcc tgcccgacga cgatgccggc gagctgcctg
     4921 ccgccgtcgt cgtgctggaa cacggcaaga ccatgaccga gaaggagatc gtggactatg
     4981 tggccagcca ggtgacaacc gccaagaagc tgcgcggcgg agtggtgttc gtggacgagg
     5041 tgcccaaggg cctgaccggc aagctggacg cccgcaagat ccgcgagatc ctgatcaagg
     5101 ctaagaaagg cggcaagatc gccgtgtaat aattctagag tcggggcggc cggccgcttc
     5161 gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     5221 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     5281 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     5341 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc
     5401 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     5461 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5521 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5581 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5641 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5701 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5761 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5821 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5881 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5941 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     6001 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     6061 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     6121 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     6181 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     6241 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     6301 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     6361 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     6421 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6481 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6541 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6601 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6661 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6721 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6781 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6841 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6901 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6961 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     7021 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     7081 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     7141 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     7201 ttgctcacat ggctcgac
//
