LOCUS       sequence-9389-ex        6629 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 6629)
  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 6629)
  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 IRF9 3'UTR
FEATURES             Location/Qualifiers
     source          1..6629
                     /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    2044..2092
                     /note="synthetic polyadenylation signal"
     promoter        2101..2852
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      2882..2891
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2888..4540
                     /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             3847..3855
                     /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    4584..4705
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4595..4614)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4649..4668
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4838..4942
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4943..5011,5012..5803)
                     /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(5161..5180)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5974..6562
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6463..6482
                     /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 cgctcgagag agcagcaggc agccattctg tccctggtgt
     1681 agagcctggg ggacccatct tccacctcac ctctttgttc ttcctgtctc ctttgaagta
     1741 gactcattct tcacacgatt gacctgtcct ctttgtgata attctcagta gttgtccgtg
     1801 ataatcgtgt cctgaaaatc ctcgcacaca ctggctggtg gagaactcaa ggctaatttt
     1861 ttatcctttt ttttttttaa ttttgagata tacgccctct ttcatctgta agggactagg
     1921 aaattccaaa tggtgtgaac ccagggggcc tttccctctt ccctgacctc ccaactctaa
     1981 agccaagcac tttatatttt cctcttagat attcactaag gacttaaagc ggccgctggc
     2041 cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaggatcta
     2101 aatgagtctt cggacctcgc gggggccgct taagcggtgg ttagggtttg tctgacgcgg
     2161 ggggaggggg aaggaacgaa acactctcat tcggaggcgg ctcggggttt ggtcttggtg
     2221 gccacgggca cgcagaagag cgccgcgatc ctcttaagca cccccccgcc ctccgtggag
     2281 gcgggggttt ggtcggcggg tggtaactgg cgggccgctg actcgggcgg gtcgcgcgcc
     2341 ccagagtgtg accttttcgg tctgctcgca gacccccggg cggcgccgcc gcggcggcga
     2401 cgggctcgct gggtcctagg ctccatgggg accgtatacg tggacaggct ctggagcatc
     2461 cgcacgactg cggtgatatt accggagacc ttctgcggga cgagccgggt cacgcggctg
     2521 acgcggagcg tccgttgggc gacaaacacc aggacggggc acaggtacac tatcttgtca
     2581 cccggaggcg cgagggactg caggagcttc agggagtggc gcagctgctt catccccgtg
     2641 gcccgttgct cgcgtttgct ggcggtgtcc ccggaagaaa tatatttgca tgtctttagt
     2701 tctatgatga cacaaacccc gcccagcgtc ttgtcattgg cgaattcgaa cacgcagatg
     2761 cagtcggggc ggcgcggtcc caggtccact tcgcatatta aggtgacgcg tgtggcctcg
     2821 aacaccgagc gaccctgcag cgacccgctt aaaagcttgg cattccggta ctgttggtaa
     2881 agccaccatg gccgatgcta agaacattaa gaagggccct gctcccttct accctctgga
     2941 ggatggcacc gctggcgagc agctgcacaa ggccatgaag aggtatgccc tggtgcctgg
     3001 caccattgcc ttcaccgatg cccacattga ggtggacatc acctatgccg agtacttcga
     3061 gatgtctgtg cgcctggccg aggccatgaa gaggtacggc ctgaacacca accaccgcat
     3121 cgtggtgtgc tctgagaact ctctgcagtt cttcatgcca gtgctgggcg ccctgttcat
     3181 cggagtggcc gtggcccctg ctaacgacat ttacaacgag cgcgagctgc tgaacagcat
     3241 gggcatttct cagcctaccg tggtgttcgt gtctaagaag ggcctgcaga agatcctgaa
     3301 cgtgcagaag aagctgccta tcatccagaa gatcatcatc atggactcta agaccgacta
     3361 ccagggcttc cagagcatgt acacattcgt gacatctcat ctgcctcctg gcttcaacga
     3421 gtacgacttc gtgccagagt ctttcgacag ggacaaaacc attgccctga tcatgaacag
     3481 ctctgggtct accggcctgc ctaagggcgt ggccctgcct catcgcaccg cctgtgtgcg
     3541 cttctctcac gcccgcgacc ctattttcgg caaccagatc atccccgaca ccgctattct
     3601 gagcgtggtg ccattccacc acggcttcgg catgttcacc accctgggct acctgatttg
     3661 cggctttcgg gtggtgctga tgtaccgctt cgaggaggag ctgttcctgc gcagcctgca
     3721 agactacaaa attcagtctg ccctgctggt gccaaccctg ttcagcttct tcgctaagag
     3781 caccctgatc gacaagtacg acctgtctaa cctgcacgag attgcctctg gcggcgcccc
     3841 actgtctaag gaggtgggcg aagccgtggc caagcgcttt catctgccag gcatccgcca
     3901 gggctacggc ctgaccgaga caaccagcgc cattctgatt accccagagg gcgacgacaa
     3961 gcctggcgcc gtgggcaagg tggtgccatt cttcgaggcc aaggtggtgg acctggacac
     4021 cggcaagacc ctgggagtga accagcgcgg cgagctgtgt gtgcgcggcc ctatgattat
     4081 gtccggctac gtgaataacc ctgaggccac aaacgccctg atcgacaagg acggctggct
     4141 gcactctggc gacattgcct actgggacga ggacgagcac ttcttcatcg tggaccgcct
     4201 gaagtctctg atcaagtaca agggctacca ggtggcccca gccgagctgg agtctatcct
     4261 gctgcagcac cctaacattt tcgacgccgg agtggccggc ctgcccgacg acgatgccgg
     4321 cgagctgcct gccgccgtcg tcgtgctgga acacggcaag accatgaccg agaaggagat
     4381 cgtggactat gtggccagcc aggtgacaac cgccaagaag ctgcgcggcg gagtggtgtt
     4441 cgtggacgag gtgcccaagg gcctgaccgg caagctggac gcccgcaaga tccgcgagat
     4501 cctgatcaag gctaagaaag gcggcaagat cgccgtgtaa taattctaga gtcggggcgg
     4561 ccggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     4621 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     4681 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     4741 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat
     4801 cgataaggat ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4861 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4921 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4981 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5041 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     5101 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     5161 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     5221 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5281 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5341 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5401 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5461 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     5521 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5581 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5641 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5701 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5761 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     5821 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5881 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5941 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6001 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6061 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6121 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6181 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6241 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6301 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6361 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6421 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6481 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6541 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6601 ttgctggcct tttgctcaca tggctcgac
//
