LOCUS       sequence-390788-        6357 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 6357)
  AUTHORS   Iwasaki S, Kawamata T, Tomari Y
  TITLE     Drosophila argonaute1 and argonaute2 employ distinct mechanisms for 
            translational repression.
  JOURNAL   Mol Cell. 2009 Apr 10;34(1):58-67. doi: 
            10.1016/j.molcel.2009.02.010. Epub 2009 Mar 5.
  PUBMED    19268617
REFERENCE   2  (bases 1 to 6357)
  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: 
            "10.1016/j.molcel.2009.02.010"; journalName: "Mol Cell"; date: 
            "2009-04-10- 10"; volume: "34"; issue: "1"; pages: "58-67"
COMMENT     Sequence Label: psiCHECK2-let-7 4x
FEATURES             Location/Qualifiers
     source          1..6357
                     /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    1772..1820
                     /note="synthetic polyadenylation signal"
     promoter        1829..2580
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      2610..2619
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2616..4268
                     /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             3575..3583
                     /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    4312..4433
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4323..4342)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4377..4396
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4566..4670
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4671..4739,4740..5531)
                     /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(4889..4908)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5702..6290
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6191..6210
                     /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 cgctcgagac tatacaagga tctacctcag tcgagactat
     1681 acaaggatct acctcagtcg agactataca aggatctacc tcagtcgaga ctatacaagg
     1741 atctacctca gtcgacgcgg ccgctggccg caataaaata tctttatttt cattacatct
     1801 gtgtgttggt tttttgtgtg aggatctaaa tgagtcttcg gacctcgcgg gggccgctta
     1861 agcggtggtt agggtttgtc tgacgcgggg ggagggggaa ggaacgaaac actctcattc
     1921 ggaggcggct cggggtttgg tcttggtggc cacgggcacg cagaagagcg ccgcgatcct
     1981 cttaagcacc cccccgccct ccgtggaggc gggggtttgg tcggcgggtg gtaactggcg
     2041 ggccgctgac tcgggcgggt cgcgcgcccc agagtgtgac cttttcggtc tgctcgcaga
     2101 cccccgggcg gcgccgccgc ggcggcgacg ggctcgctgg gtcctaggct ccatggggac
     2161 cgtatacgtg gacaggctct ggagcatccg cacgactgcg gtgatattac cggagacctt
     2221 ctgcgggacg agccgggtca cgcggctgac gcggagcgtc cgttgggcga caaacaccag
     2281 gacggggcac aggtacacta tcttgtcacc cggaggcgcg agggactgca ggagcttcag
     2341 ggagtggcgc agctgcttca tccccgtggc ccgttgctcg cgtttgctgg cggtgtcccc
     2401 ggaagaaata tatttgcatg tctttagttc tatgatgaca caaaccccgc ccagcgtctt
     2461 gtcattggcg aattcgaaca cgcagatgca gtcggggcgg cgcggtccca ggtccacttc
     2521 gcatattaag gtgacgcgtg tggcctcgaa caccgagcga ccctgcagcg acccgcttaa
     2581 aagcttggca ttccggtact gttggtaaag ccaccatggc cgatgctaag aacattaaga
     2641 agggccctgc tcccttctac cctctggagg atggcaccgc tggcgagcag ctgcacaagg
     2701 ccatgaagag gtatgccctg gtgcctggca ccattgcctt caccgatgcc cacattgagg
     2761 tggacatcac ctatgccgag tacttcgaga tgtctgtgcg cctggccgag gccatgaaga
     2821 ggtacggcct gaacaccaac caccgcatcg tggtgtgctc tgagaactct ctgcagttct
     2881 tcatgccagt gctgggcgcc ctgttcatcg gagtggccgt ggcccctgct aacgacattt
     2941 acaacgagcg cgagctgctg aacagcatgg gcatttctca gcctaccgtg gtgttcgtgt
     3001 ctaagaaggg cctgcagaag atcctgaacg tgcagaagaa gctgcctatc atccagaaga
     3061 tcatcatcat ggactctaag accgactacc agggcttcca gagcatgtac acattcgtga
     3121 catctcatct gcctcctggc ttcaacgagt acgacttcgt gccagagtct ttcgacaggg
     3181 acaaaaccat tgccctgatc atgaacagct ctgggtctac cggcctgcct aagggcgtgg
     3241 ccctgcctca tcgcaccgcc tgtgtgcgct tctctcacgc ccgcgaccct attttcggca
     3301 accagatcat ccccgacacc gctattctga gcgtggtgcc attccaccac ggcttcggca
     3361 tgttcaccac cctgggctac ctgatttgcg gctttcgggt ggtgctgatg taccgcttcg
     3421 aggaggagct gttcctgcgc agcctgcaag actacaaaat tcagtctgcc ctgctggtgc
     3481 caaccctgtt cagcttcttc gctaagagca ccctgatcga caagtacgac ctgtctaacc
     3541 tgcacgagat tgcctctggc ggcgccccac tgtctaagga ggtgggcgaa gccgtggcca
     3601 agcgctttca tctgccaggc atccgccagg gctacggcct gaccgagaca accagcgcca
     3661 ttctgattac cccagagggc gacgacaagc ctggcgccgt gggcaaggtg gtgccattct
     3721 tcgaggccaa ggtggtggac ctggacaccg gcaagaccct gggagtgaac cagcgcggcg
     3781 agctgtgtgt gcgcggccct atgattatgt ccggctacgt gaataaccct gaggccacaa
     3841 acgccctgat cgacaaggac ggctggctgc actctggcga cattgcctac tgggacgagg
     3901 acgagcactt cttcatcgtg gaccgcctga agtctctgat caagtacaag ggctaccagg
     3961 tggccccagc cgagctggag tctatcctgc tgcagcaccc taacattttc gacgccggag
     4021 tggccggcct gcccgacgac gatgccggcg agctgcctgc cgccgtcgtc gtgctggaac
     4081 acggcaagac catgaccgag aaggagatcg tggactatgt ggccagccag gtgacaaccg
     4141 ccaagaagct gcgcggcgga gtggtgttcg tggacgaggt gcccaagggc ctgaccggca
     4201 agctggacgc ccgcaagatc cgcgagatcc tgatcaaggc taagaaaggc ggcaagatcg
     4261 ccgtgtaata attctagagt cggggcggcc ggccgcttcg agcagacatg ataagataca
     4321 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     4381 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     4441 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     4501 agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc aggtggcact tttcggggaa
     4561 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4621 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4681 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4741 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4801 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4861 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4921 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4981 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5041 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5101 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5161 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5221 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5281 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5341 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5401 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5461 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5521 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5581 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5641 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5701 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5761 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5821 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     5881 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5941 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6001 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6061 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6121 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6181 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6241 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6301 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg gctcgac
//
