LOCUS       sequence-9968-ex        8108 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 8108)
  AUTHORS   Eulalio A, Behm-Ansmant I, Schweizer D, Izaurralde E
  TITLE     P-body formation is a consequence, not the cause, of RNA-mediated 
            gene silencing.
  JOURNAL   Mol Cell Biol. 2007 Jun . 27(11):3970-81.
  PUBMED    17403906
REFERENCE   2  (bases 1 to 8108)
  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: "Mol Cell 
            Biol. 2007 Jun . 27(11):3970-81."
COMMENT     Sequence Label: pAc5.1B-EGFP-DmEDC3
FEATURES             Location/Qualifiers
     source          1..8108
                     /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             5393..5434
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5444..5461
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5484..5501)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5534..5668
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5558..5577)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5612..5631
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6290..6878)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6370..6389)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7049..7840,7841..7909))
                     /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     7672..7691
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7910..8014)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8082..8100
                     /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 ttgatgggtc cgacggatca agactggatc ggctgtgcag tgtctatagc
     3361 ctgcgacgag gtcctgggcg tcttccaggg cctgatcaag cagatttctg ccgaggagat
     3421 caccattgtg agggctttcc gcaatggcgt gccgctgcga aagcagaacg ccgaggtggt
     3481 cctcaagtgc acggacattc gcagcatcga tctcatcgag cccgcaaagc aagatctgga
     3541 tgggcacacg gcgccgccgc cagtggtcaa caagcccacg cccgtcaagc tgccccactt
     3601 ctccaacatc ctgggcaagc agcagcagct ccaactccag cagcagcagc agcagctaca
     3661 actccagcag caacagaaac agcagttcca ggagcaggaa cgagaacagg atctgccatc
     3721 gacgccgcgg tcaagggcca acgacaatgg acgggtagcg gctggcggag catcgagttc
     3781 ttcgggccca cgtggcaact tcaattcggc gctaagcgat aagatgcatc agctgaagct
     3841 aatcgagacc aacggctcaa atggcaccct tagaacacca caaacatccc gtgcgtccag
     3901 cgctcagcag cagccgcaga tatcaacgac tcccaacagc gtggccgcct tttttggcaa
     3961 catgataccg ccgaaggtcg aggtgaaact gggctcctac gtgagcaaca cccgggagag
     4021 ttactgcagc agcagcggag atagcggcga ggcaactggt ttgtcgttgg gcagcagcaa
     4081 acccatagat atcgtcagca acggagatgg attttacaag cagacggcag cctcgagcta
     4141 tgggaataca aatggaaatg taagaaggaa tggcaatgcc aataacaatg gaaatggcac
     4201 cggaaatggt tcttatacca atggaaatgg caacggcaac gggaagaaca agcggaacag
     4261 ggtgcgaagg gagagcagca tgcggcagca acaggtgcaa ctgacttttg gcagcgaggc
     4321 ggacgatccg ctgatccacg aggatttcga ttttgagggc aatctagcgc tgttcgacaa
     4381 gcaggcaatt tgggacgaca tcgagtcgac gacccaaaaa ccagatgtgg tgcggcacat
     4441 tgtgaacaat catcatcata agccagagca gaagtatcgg cacgacgaga acatcctggc
     4501 cagcaaacca ctacaattgc gccagatcga aagcatgttc ggcggcagcc aggactttgt
     4561 gacggacgac ggtctcatta taccgacgat tccggcttac gttcggaata aaatcgagat
     4621 aagtgccgat aaggcgggcc tctctctgca gaggcagatc gacattttgg cgcgcggagc
     4681 cagtgatctg gccattacac tgctgggcgg cgccaggcga ttgacgccgg ccaacaacca
     4741 tcagtggccg aagattgcca tcatctgcga tggcggcaaa aatatgagaa caattaacat
     4801 tggagcagcc acaggtcgcc agctggcttc gcatgggctt actgtcctgc tgtacgtgga
     4861 gcaggccaaa ctgctggagc agaacagcag cagcccagag atttcattgt tcaaggcaac
     4921 cgacaatgtc atagttcact cagtagacgc cttacccacg ccagatcttg tgatattatc
     4981 aactaacacc gctaatttat cggatgcgat caggaaatgg ctcagtgtaa atcgcgcttc
     5041 ggtgcttgcc atagacccgc cgccgtgtgg cattaatgag gttgctatca agtactcaat
     5101 actacccata ctaccattaa atggcatatc aacagcaaca acatcatcat cctcttcggc
     5161 tgccgccaca ccaactccca ttgctagcac ttcggcagca gcatcggcta ccaaatcagc
     5221 agcatcaacg aacaattgtg gcaaattgta tttatgtaat ctaggtatac cggataaatt
     5281 ctatcgtgat tgtggcatta agtacaaaag tccgtatgga cataaatatg tgataccgat
     5341 tcactcaaag gactgaggcg gccgctcgag tctagagggc ccgcggttcg aaggtaagcc
     5401 tatccctaac cctctcctcg gtctcgattc tacgcgtacc ggtcatcatc accatcacca
     5461 ttgagtttaa acccgctgat cagcctcgac tgtgccttct aaggcctgag ctcgctgatc
     5521 agcctcgatc gaggatccag acatgataag atacattgat gagtttggac aaaccacaac
     5581 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     5641 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     5701 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
     5761 ggctgattat gatcagtcga ccgatgccct tgagagcctt caacccagtc agctccttcc
     5821 ggtgggcgcg gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac
     5881 tcgtaggaca ggtgccggca gcgctctggg tcattttcgg cgaggaccgc tttcgctgga
     5941 gcgcgacgat gatcggcctg tcgcttgcgg tattcggaat cttgcacgcc ctcgctcaag
     6001 ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa gcaggccatt atcgccggca
     6061 tggcggccga cgcgctgggc tacgtcttgc tggcgttcgc gacgcgaggc tggatggcct
     6121 tccccattat gattcttctc gcttccggcg gcatcgggat gcccgcgttg caggccatgc
     6181 tgtccaggca ggtagatgac gaccatcagg gacagcttca aggatcgctc gcggctctta
     6241 ccagccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6301 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6361 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6421 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6481 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6541 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6601 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6661 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6721 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6781 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6841 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6901 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6961 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     7021 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     7081 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     7141 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7201 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7261 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7321 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt
     7381 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7441 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7501 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7561 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7621 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg
     7681 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7741 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7801 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7861 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7921 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7981 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     8041 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     8101 cctttcgt
//
