LOCUS       sequence-10419-e        9018 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 9018)
  AUTHORS   Eulalio A, Huntzinger E, Izaurralde E
  TITLE     GW182 interaction with Argonaute is essential for miRNA-mediated 
            translational repression and mRNA decay.
  JOURNAL   Nat Struct Mol Biol. 2008 Apr . 15(4):346-53.
  PUBMED    18345015
REFERENCE   2  (bases 1 to 9018)
  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"; journalName: "Nat Struct 
            Mol Biol. 2008 Apr . 15(4):346-53."
COMMENT     Sequence Label: pAc5.1B-EGFP-DmAGO1
FEATURES             Location/Qualifiers
     source          1..9018
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..2568
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2588..2590,2591..2593,2594..3304)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2588..2590,2591..2593,2594..3304)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2633..2654)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2894..2913)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3241..3262
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6303..6344
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6354..6371
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6394..6411)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6444..6578
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6468..6487)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6522..6541
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7200..7788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7280..7299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7959..8750,8751..8819))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8582..8601
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8820..8924)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8992..9010
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cttcaaggaa ttaattctat attctaaaaa cacaaatgat acttctaaaa aaaatcatga
       61 atggcatcaa ctctgaatca aatctttgca gatgcaccta cttctcattt ccactgtcac
      121 atcatttttc cagatctcgc tgcctgttat gtggcccaca aaccaagaca cgttttatgg
      181 ccattaaagc tggctgatcg tcgccaaaca ccaaatacat atcaatatgt acattcgaga
      241 aagaagcgat caaagaagcg tcttcgggcg agtaggagaa tgcggaggag aaggagaacg
      301 agctgatcta gtatctctcc acaatccaat gccaactgac caactggcca tattcggagc
      361 aatttgaagc caatttccat cgcctggcga tcgctccatt cttggctata tgtttttcac
      421 cgttcccggg gccattttca aagactcgtc ggtaagataa gattgtgtca ctcgctgtct
      481 ctcttcattt gtcgaagaat gctgaggaat ttcgcgatga cgtcggcgag tattttgaag
      541 aatgagaata atttgtattt atacgaaaat cagttagtgg aattttctac aaaaacatgt
      601 tatctataga taattttgtt gcaaaatatg ttgactatga caaagattgt atgtatatac
      661 ctttaatgta ttctcatttt cttatgtatt tataatggca atgatgatac tgatgatatt
      721 ttaagatgat gccagaccac aggctgattt ctgcgtcttt tgccgaacgc agtgcatgtg
      781 cggttgttgt tttttggaat agtttcaatt ttcggactgt ccgctttgat ttcagtttct
      841 tggcttattc aaaaagcaaa gtaaagccaa aaaagcgaga tggcaatacc aaatgcggca
      901 aaacggtagt ggaaggaaag gggtgcgggg cagcggaagg aagggtgggg cggggcgtgg
      961 cggggtctgt ggctgggcgc gacgtcaccg acgttggagc cactcctttg accatgtgtg
     1021 cgtgtgtgta ttattcgtgt ctcgccactc gccggttgtt tttttctttt tatctcgctc
     1081 tctctagcgc catctcgtac gcatgctcaa cgcaccgcat gttgccgtgt cctttatgcg
     1141 tcattttggc tcgaaatagg caattattta aacaaagatt agtcaacgaa aacgctaaaa
     1201 taaataagtc tacaatatgg ttacttattg ccatgtgtgt gcagccaacg atagcaacaa
     1261 aagcaacaac acagtggctt tccctctttc actttttgtt tgcaagcgcg tgcgagcaag
     1321 acggcacgac cggcaaacgc aattacgctg acaaagagca gacgaagttt tggccgaaaa
     1381 acatcaaggc gcctgatacg aatgcatttg caataacaat tgcgatattt aatattgttt
     1441 atgaagctgt ttgacttcaa aacacacaaa aaaaaaaata aaacaaatta tttgaaagag
     1501 aattaggaat cggacagctt atcgttacgg gctaacagca caccgagacg aaatagctta
     1561 cctgacgtca cagcctctgg aagaactgcc gccaagcaga gagagagaga aaaagaggga
     1621 gagcagctta gaccgcatgt gcttgtgtgt gaggcgtctc tctcttcgtc tcctgtttgc
     1681 gcaaacgcat agactgcact gagaaaatcg attacctatt ttttatgaat gaatatttgc
     1741 actattacta ttcaaaacta ttaagatagc aatcacattc aatagccaaa tactatacca
     1801 cctgagcgat gcaacgaaat gatcaatttg agcaaaaatg ctgcatattt aggacggcat
     1861 cattatagaa atgcttcttg ctgtgtactt ttctctcgtc tggcagctgt ttcgccgtta
     1921 ttgttaaaac cggcttaagt taggtgtgtt ttctacgact agtgatgccc ctactagaag
     1981 atgtgtgttg cacaaatgtc cctgaataac caatttgaag tgcagatagc agtaaacgta
     2041 agctaatatg aatattattt aactgtaatg ttttaatatc gctggacatt actaataaac
     2101 ccactataaa cacatgtaca tatgtatgtt ttggcataca atgagtagtt ggggaaaaaa
     2161 tgtgtaaaag caccgtgacc atcacagcat aaagataacc agctgaagta tcgaatatga
     2221 gtaaccccca aattgaatca catgccgcaa ctgataggac ccatggaagt acactcttca
     2281 tggcgatata caagacacac acaagcacga acacccagtt gcggaggaaa ttctccgtaa
     2341 atgaaaaccc aatcggcgaa caattcatac ccatatatgg taaaagtttt gaacgcgact
     2401 tgagagcgga gagcattgcg gctgataagg ttttagcgct aagcgggctt tataaaacgg
     2461 gctgcgggac cagttttcat atcactaccg tttgagttct tgtgctgtgt ggatactcct
     2521 cccgacacaa agccgctcca tcagccagca gtcgtctaat ccagagaccc cggatcgggg
     2581 taccaacatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     2641 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     2701 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     2761 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     2821 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     2881 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     2941 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     3001 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     3061 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     3121 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     3181 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     3241 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     3301 caagccaagc ttgaattcta tgtccacgga gcgtgagctg gctcccggtg ggccagctca
     3361 gctccatccg cacacgctgc cgctgacttt tccggatctc cagatgacct ccaccgtggg
     3421 catcattggg aaagtctacg agtcacagtg gaccccctcg cccactcggc ctcagagtcc
     3481 ctctcaggcg cagactagct tcgacacgct cacatcacca ccagcgcccg gctcgtcggt
     3541 caatcccacc gccgtcacca gcccaagtgc ccagaatgtg gccgctggtg gagcaactgt
     3601 ggccggtgca gcagcaactg ccgcccaggt ggcctccgcc ttgggtgcca ccaccggcag
     3661 cgtgacgcca gcaattgcca ccgcaacgcc agccacgcag ccggatatgc ctgtctttac
     3721 gtgtccacga cgcccgaatc tcggacgaga gggtcgcccg attgtgctgc gcgccaatca
     3781 cttccaggtg acaatgccgc gtggctatgt gcatcactat gacatcaata tacagccgga
     3841 caagtgtccg cgaaaggtga accgtgagat tatcgagact atggtgcatg cctatagcaa
     3901 gatcttcgga gtgctcaagc cggtgttcga tggtcgcaac aatctgtata cccgcgatcc
     3961 cctgcccatt ggcaacgagc gtctagagct ggaggttact ctacccggcg agggcaaaga
     4021 tcgaatcttt cgcgtaacga tcaagtggca ggctcaggtc tcgctcttca atttggagga
     4081 agctctcgaa ggccgcacgc ggcagatacc ctatgatgcc attttggcgc tcgatgtggt
     4141 catgcgccat ctgcccagca tgacgtacac gccagtggga cgtagcttct tcagttcccc
     4201 ggagggttac taccatcccc tgggtggtgg acgcgaggtt tggttcggtt tccatcagag
     4261 cgtaaggccc tcgcagtgga agatgatgct caatatcgat gtctcggcca ccgctttcta
     4321 caaggctcaa ccagtcattg acttcatgtg cgaggtgctg gacattcgcg acatcaacga
     4381 gcagcgcaaa ccgctcaccg attcgcagcg cgtcaagttc accaaagaaa ttaaaggcct
     4441 gaagatcgag atcacccact gcggccagat gcgtcgcaag tatcgtgtgt gcaacgtcac
     4501 tcgccgcccc gctcagatgc aatcattccc actgcagctg gagaacggac agaccgtaga
     4561 gtgcaccgtg gccaagtact tcctggacaa gtaccgcatg aaattgcgct acccgcactt
     4621 gccctgcctg caggttggcc aagagcacaa gcacacttac ctgcctctag aggtgtgcaa
     4681 cattgtggcc ggacagcggt gcattaaaaa gctgaccgat atgcagacgt cgaccatgat
     4741 caaggccaca gctcgttcag ctccggatcg tgagcgtgag attaacaact tggtaaagcg
     4801 cgccgacttc aacaacgatt cgtatgtgca agagtttggc ctgaccatct ccaattcgat
     4861 gatggaggta cgaggacgcg tcttgcctcc tcccaagctt cagtatgggg gacgtgtgtc
     4921 taccggcctc accggccagc agctgttccc gccacagaac aaggtgagct tggcctcgcc
     4981 caaccagggt gtatgggata tgcgaggcaa gcagttcttc actggcgtcg agatccgcat
     5041 ctgggccatc gcctgttttg ccccacagcg cacggtgcgc gaggatgcgc tgcgtaattt
     5101 cacccagcag ctgcagaaga tctcaaacga tgcaggcatg ccgataattg gacagccgtg
     5161 cttctgtaag tacgccaccg ggccggatca agtggaaccc atgttccgtt acctgaagat
     5221 caccttcccc ggcctgcaac tcgtcgtggt tgtgctgccc ggcaagactc cagtatacgc
     5281 cgaggtgaag cgtgtaggtg acaccgttct gggtatggcc acccagtgtg tgcaggccaa
     5341 gaacgtgaac aagacatcgc cacagacgct ctctaatctg tgtctgaaga tcaacgtcaa
     5401 gttgggcggc atcaattcaa ttttggtccc ctccattcgg ccaaaggtct tcaatgagcc
     5461 ggtcatcttt ttgggtgccg atgtgacaca cccaccagct ggcgacaaca agaaaccatc
     5521 gattgccgcc gtcgtgggct ccatggatgc ccatccatcg cggtatgccg ccaccgttcg
     5581 cgtacagcag caccgacagg agatcatcca ggagctgagc agcatggtgc gcgagctgtt
     5641 gatcatgttc tacaagtcga cgggcggcta caagccccac cgcatcatac tctatcgtga
     5701 cggagtctcc gagggacaat tcccacatgt cctgcaacac gaattgaccg ccattcgcga
     5761 ggcctgcatt aagctagagc cagagtatcg gccgggcatc acattcattg tggtgcagaa
     5821 gcgccatcac actcgactct tctgcgcgga gaagaaggag cagagcggca aatcgggcaa
     5881 tattcccgca ggcaccaccg tcgatgtggg catcacacat cccaccgaat ttgatttcta
     5941 tctatgcagc catcagggca tccagggcac cagtcgcccc tcgcactacc acgttctgtg
     6001 ggacgacaat cactttgact cggacgagct gcagtgcctc acgtatcagc tatgccatac
     6061 gtacgtgcgc tgcacccgat ccgtcagtat accggcgcca gcctactacg cccatttagt
     6121 ggccttccgt gccagatatc atctggtgga gaaggagcac gattcgggcg agggttcgca
     6181 ccagagcggc tgctcagagg atcgtacgcc aggtgccatg gccagggcca tcactgtgca
     6241 cgcggatacc aagaaggtca tgtactttgc ctaactcgag tctagagggc ccgcggttcg
     6301 aaggtaagcc tatccctaac cctctcctcg gtctcgattc tacgcgtacc ggtcatcatc
     6361 accatcacca ttgagtttaa acccgctgat cagcctcgac tgtgccttct aaggcctgag
     6421 ctcgctgatc agcctcgatc gaggatccag acatgataag atacattgat gagtttggac
     6481 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     6541 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     6601 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     6661 aatgtggtat ggctgattat gatcagtcga ccgatgccct tgagagcctt caacccagtc
     6721 agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac tgtcttcttt
     6781 atcatgcaac tcgtaggaca ggtgccggca gcgctctggg tcattttcgg cgaggaccgc
     6841 tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg tattcggaat cttgcacgcc
     6901 ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa gcaggccatt
     6961 atcgccggca tggcggccga cgcgctgggc tacgtcttgc tggcgttcgc gacgcgaggc
     7021 tggatggcct tccccattat gattcttctc gcttccggcg gcatcgggat gcccgcgttg
     7081 caggccatgc tgtccaggca ggtagatgac gaccatcagg gacagcttca aggatcgctc
     7141 gcggctctta ccagccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     7201 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     7261 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     7321 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     7381 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     7441 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     7501 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     7561 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     7621 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     7681 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     7741 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     7801 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     7861 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     7921 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     7981 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     8041 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     8101 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     8161 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     8221 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc
     8281 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     8341 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     8401 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     8461 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     8521 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg
     8581 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     8641 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     8701 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     8761 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     8821 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     8881 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     8941 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     9001 atcacgaggc cctttcgt
//
