LOCUS       sequence-313838-        6465 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 6465)
  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 6465)
  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 8x
FEATURES             Location/Qualifiers
     source          1..6465
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    237..285
                     /note="synthetic polyadenylation signal"
     promoter        294..1045
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      1075..1084
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1081..2733
                     /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             2040..2048
                     /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    2777..2898
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2788..2807)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2842..2861
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3031..3135
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3136..3204,3205..3996)
                     /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(3354..3373)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4167..4755
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4656..4675
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4884..5241
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4884..5075
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4907..4927)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5092..5227
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          5311..5443
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     5426..5445
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        5488..5506
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      5510..5519
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5516..6451
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
ORIGIN
        1 tcgagactat acaaggatct acctcagtcg agactataca aggatctacc tcagtcgaga
       61 ctatacaagg atctacctca gtcgagacta tacaaggatc tacctcagtc gagactatac
      121 aaggatctac ctcagtcgag actatacaag gatctacctc agtcgagact atacaaggat
      181 ctacctcagt cgagactata caaggatcta cctcagtcga cgcggccgct ggccgcaata
      241 aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaggat ctaaatgagt
      301 cttcggacct cgcgggggcc gcttaagcgg tggttagggt ttgtctgacg cggggggagg
      361 gggaaggaac gaaacactct cattcggagg cggctcgggg tttggtcttg gtggccacgg
      421 gcacgcagaa gagcgccgcg atcctcttaa gcaccccccc gccctccgtg gaggcggggg
      481 tttggtcggc gggtggtaac tggcgggccg ctgactcggg cgggtcgcgc gccccagagt
      541 gtgacctttt cggtctgctc gcagaccccc gggcggcgcc gccgcggcgg cgacgggctc
      601 gctgggtcct aggctccatg gggaccgtat acgtggacag gctctggagc atccgcacga
      661 ctgcggtgat attaccggag accttctgcg ggacgagccg ggtcacgcgg ctgacgcgga
      721 gcgtccgttg ggcgacaaac accaggacgg ggcacaggta cactatcttg tcacccggag
      781 gcgcgaggga ctgcaggagc ttcagggagt ggcgcagctg cttcatcccc gtggcccgtt
      841 gctcgcgttt gctggcggtg tccccggaag aaatatattt gcatgtcttt agttctatga
      901 tgacacaaac cccgcccagc gtcttgtcat tggcgaattc gaacacgcag atgcagtcgg
      961 ggcggcgcgg tcccaggtcc acttcgcata ttaaggtgac gcgtgtggcc tcgaacaccg
     1021 agcgaccctg cagcgacccg cttaaaagct tggcattccg gtactgttgg taaagccacc
     1081 atggccgatg ctaagaacat taagaagggc cctgctccct tctaccctct ggaggatggc
     1141 accgctggcg agcagctgca caaggccatg aagaggtatg ccctggtgcc tggcaccatt
     1201 gccttcaccg atgcccacat tgaggtggac atcacctatg ccgagtactt cgagatgtct
     1261 gtgcgcctgg ccgaggccat gaagaggtac ggcctgaaca ccaaccaccg catcgtggtg
     1321 tgctctgaga actctctgca gttcttcatg ccagtgctgg gcgccctgtt catcggagtg
     1381 gccgtggccc ctgctaacga catttacaac gagcgcgagc tgctgaacag catgggcatt
     1441 tctcagccta ccgtggtgtt cgtgtctaag aagggcctgc agaagatcct gaacgtgcag
     1501 aagaagctgc ctatcatcca gaagatcatc atcatggact ctaagaccga ctaccagggc
     1561 ttccagagca tgtacacatt cgtgacatct catctgcctc ctggcttcaa cgagtacgac
     1621 ttcgtgccag agtctttcga cagggacaaa accattgccc tgatcatgaa cagctctggg
     1681 tctaccggcc tgcctaaggg cgtggccctg cctcatcgca ccgcctgtgt gcgcttctct
     1741 cacgcccgcg accctatttt cggcaaccag atcatccccg acaccgctat tctgagcgtg
     1801 gtgccattcc accacggctt cggcatgttc accaccctgg gctacctgat ttgcggcttt
     1861 cgggtggtgc tgatgtaccg cttcgaggag gagctgttcc tgcgcagcct gcaagactac
     1921 aaaattcagt ctgccctgct ggtgccaacc ctgttcagct tcttcgctaa gagcaccctg
     1981 atcgacaagt acgacctgtc taacctgcac gagattgcct ctggcggcgc cccactgtct
     2041 aaggaggtgg gcgaagccgt ggccaagcgc tttcatctgc caggcatccg ccagggctac
     2101 ggcctgaccg agacaaccag cgccattctg attaccccag agggcgacga caagcctggc
     2161 gccgtgggca aggtggtgcc attcttcgag gccaaggtgg tggacctgga caccggcaag
     2221 accctgggag tgaaccagcg cggcgagctg tgtgtgcgcg gccctatgat tatgtccggc
     2281 tacgtgaata accctgaggc cacaaacgcc ctgatcgaca aggacggctg gctgcactct
     2341 ggcgacattg cctactggga cgaggacgag cacttcttca tcgtggaccg cctgaagtct
     2401 ctgatcaagt acaagggcta ccaggtggcc ccagccgagc tggagtctat cctgctgcag
     2461 caccctaaca ttttcgacgc cggagtggcc ggcctgcccg acgacgatgc cggcgagctg
     2521 cctgccgccg tcgtcgtgct ggaacacggc aagaccatga ccgagaagga gatcgtggac
     2581 tatgtggcca gccaggtgac aaccgccaag aagctgcgcg gcggagtggt gttcgtggac
     2641 gaggtgccca agggcctgac cggcaagctg gacgcccgca agatccgcga gatcctgatc
     2701 aaggctaaga aaggcggcaa gatcgccgtg taataattct agagtcgggg cggccggccg
     2761 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     2821 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     2881 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     2941 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag
     3001 gatccaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     3061 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     3121 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     3181 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     3241 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     3301 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     3361 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     3421 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     3481 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     3541 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     3601 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     3661 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     3721 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     3781 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     3841 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     3901 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     3961 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     4021 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4081 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4141 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4201 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4261 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     4321 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4381 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4441 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4501 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4561 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4621 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     4681 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4741 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     4801 ccttttgctc acatggctcg acagatctgc gcagcaccat ggcctgaaat aacctctgaa
     4861 agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     4921 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4981 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5041 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     5101 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     5161 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     5221 ggaggcctag gcttttgcaa aaagcttgat tcttctgaca caacagtctc gaacttaagc
     5281 tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa
     5341 ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca
     5401 cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc
     5461 aattacagct cttaaggcta gagtacttaa tacgactcac tataggctag ccaccatggc
     5521 ttccaaggtg tacgaccccg agcaacgcaa acgcatgatc actgggcctc agtggtgggc
     5581 tcgctgcaag caaatgaacg tgctggactc cttcatcaac tactatgatt ccgagaagca
     5641 cgccgagaac gccgtgattt ttctgcatgg taacgctgcc tccagctacc tgtggaggca
     5701 cgtcgtgcct cacatcgagc ccgtggctag atgcatcatc cctgatctga tcggaatggg
     5761 taagtccggc aagagcggga atggctcata tcgcctcctg gatcactaca agtacctcac
     5821 cgcttggttc gagctgctga accttccaaa gaaaatcatc tttgtgggcc acgactgggg
     5881 ggcttgtctg gcctttcact actcctacga gcaccaagac aagatcaagg ccatcgtcca
     5941 tgctgagagt gtcgtggacg tgatcgagtc ctgggacgag tggcctgaca tcgaggagga
     6001 tatcgccctg atcaagagcg aagagggcga gaaaatggtg cttgagaata acttcttcgt
     6061 cgagaccatg ctcccaagca agatcatgcg gaaactggag cctgaggagt tcgctgccta
     6121 cctggagcca ttcaaggaga agggcgaggt tagacggcct accctctcct ggcctcgcga
     6181 gatccctctc gttaagggag gcaagcccga cgtcgtccag attgtccgca actacaacgc
     6241 ctaccttcgg gccagcgacg atctgcctaa gatgttcatc gagtccgacc ctgggttctt
     6301 ttccaacgct attgtcgagg gagctaagaa gttccctaac accgagttcg tgaaggtgaa
     6361 gggcctccac ttcagccagg aggacgctcc agatgaaatg ggtaagtaca tcaagagctt
     6421 cgtggagcgc gtgctgaaga acgagcagta attctaggcg atcgc
//
