LOCUS       sequence-315753-        4443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression construct with ubiquitous expression to test fluorescence
            level on microscope.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4443)
  AUTHORS   Aljohani MD, El Mouridi S, Priyadarshini M, Vargas-Velazquez AM, 
            Frokjaer-Jensen C
  TITLE     Engineering rules that minimize germline silencing of transgenes in 
            simple extrachromosomal arrays in C. elegans.
  JOURNAL   Nat Commun. 2020 Dec 9;11(1):6300. doi: 10.1038/s41467-020-19898-0.
  PUBMED    33298957
REFERENCE   2  (bases 1 to 4443)
  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 
            Commun."; date: "2020-12-9"; volume: "11(1):6300. doi"; pages: " 
            10.1038/s41467-020-19898-0"
COMMENT     Sequence Label: pMDJ17 - pEXP[Peft-3 | dendra2 (NLS) | tbb-2 UTR]
FEATURES             Location/Qualifiers
     source          1..4443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1..17
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    37..57
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        58..657
                     /gene="eft-3"
                     /label=eft-3 promoter
                     /note="promoter for the C. elegans elongation factor 
                     1-alpha gene (Mitrovich and Anderson, 2000)"
     protein_bind    683..707
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             716..736
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1621..1692
                     /codon_start=1
                     /product="nuclear localization signal from the C. elegans 
                     EGL-13 transcription factor (Lyssenko et al., 2007)"
                     /label=EGL-13 NLS
                     /translation="SRRRKANPTKLSENAKKLAKEVEN"
     protein_bind    complement(1700..1724)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      1729..2052
                     /gene="tbb-2"
                     /label=tbb-2 terminator
                     /note="transcription terminator for the C. elegans tubulin 
                     beta-2 gene"
     protein_bind    complement(2056..2076)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(2084..2101)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2084..2100)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2093..2115)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2428..2450
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2488..2506)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2574..2678
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2679..2747,2748..3539)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGSDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2897..2916)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3710..4298
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4199..4218
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(4425..4443,1..4)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 caggaaacag ctatgaccat gattacgcca agctatcaac tttgtataga aaagttggca
       61 cctttggtct tttattgtca acttccattg gttcttccat tgtttctgtt aaattaatga
      121 atttttcata aaataaagac attatacaat ataaaaatga agaatttatt gaaaataaac
      181 tgccagagag aaaaagtatg caacactccc gccgagagtg tttgaaatgg tgtacggtac
      241 attttcgtgc taggagttag atgtgcaggc agcaacgaga gggggagaga tttttttggg
      301 ccttgtgaaa ttaacgtgag ttttctggtc atctgactaa tcatgttggt tttttgttgg
      361 tttattttgt ttttatcttt gtttttatcc agattaggaa atttaaattt tatgaattta
      421 taatgaggtc aaacattcag tcccagcgtt tttcctgttc tcactgttta gtcgaatttt
      481 tattttaggc tttcaacaaa tgttctaact gtcttatttg tgacctcact ttttatattt
      541 ttttaatttt taaaaatatt agaagtttct aggataattt tttcgacttt tattctctct
      601 accgtccgca ctcttcttac ttttaaatta aattgttttt ttttcagttg ggaaacactt
      661 tgctcactcc gtagcagcca tggcaagttt gtacaaaaaa gcaggctcgg gatccccaaa
      721 gaagaagcgt aaggtcaaca ccccaggtaa cacttagttt ttgttgctct gtttaaaaat
      781 taattttaag gaatcaacct catcaaggag gacatgcgtg tcaaggtcca catggaggga
      841 aacgtcaacg gacacgcctt cgtcatcgag ggagagggaa agggaaagcc atacgaggga
      901 acccaaaccg ccaacctcac cgtcaaggta agttttcttg agacctttac taactaacta
      961 ggtctcatag attttcagga gggagcccca ctcccattct cctacgacat cctcaccacc
     1021 gccgtccact acggaaaccg tgtcttcacc aagtacccag aggacatccc agactacttc
     1081 aagcaatcct tcccagaggg atactcctgg gagcgtacca tgaccttcga ggacaaggga
     1141 atctgcacca tccgttccga catctccctc gagggagact gcttcttcca aaacgtccgt
     1201 ttcaaggtaa gttatgtaga gacctttact aactaactag gtctcaaaca ttttcaggga
     1261 accaacttcc caccaaacgg accagtcatg caaaagaaga ccctcaagtg ggagccatcc
     1321 accgagaagc tccacgtccg tgacggactc ctcgtcggaa acatcaacat ggccctcctc
     1381 ctcgagggag gaggacacta cctctgcgac ttcaaggtaa gttctttaga gacctttact
     1441 aactaactag gtctcagtaa ttttcagacc acctacaagg ccaagaaggt cgtccaactc
     1501 ccagacgccc acttcgtcga ccaccgtatc gagatcctcg gaaacgactc cgactacaac
     1561 aaggtcaagc tctacgagca cgccgtcgcc cgttactccc cactcccatc ccaagtctgg
     1621 tcccgtcgtc gtaaggccaa cccaaccaag ctctccgaga acgccaagaa gctcgccaag
     1681 gaggtcgaga acggatccaa cccagctttc ttgtacaaag tgggataaat gcaaaatcct
     1741 ttcaagcatt cccttcttct ctatcactct tctttctttt tgtcaaaaaa ttctctcgct
     1801 aatttatttg cttttttaat gttattattt tatgactttt tatagtcact gaaaagtttg
     1861 catctgagtg aagtgaatgc tatcaaaatg tgattctgtc tgatgtactt tcacaatctc
     1921 tcttcaattc cattttgaag tgctttaaac ccgaaaggtt gagaaaaatg cgagcgctca
     1981 aatatttgta ttgtgttcgt tgagtgaccc aacaaaaaga ggaaacttta ttgtgccgcc
     2041 aagaaaaaag tctcacaact ttattataca tagttgataa ttcactggcc gtcgttttac
     2101 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     2161 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     2221 gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     2281 tttcacaccg catatggtgc actctcagtg caatctgctc tgatgccgca tagttaagcc
     2341 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     2401 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     2461 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     2521 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     2581 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2641 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     2701 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     2761 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     2821 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     2881 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     2941 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     3001 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     3061 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     3121 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     3181 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     3241 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     3301 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     3361 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     3421 ggtcagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     3481 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     3541 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     3601 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     3661 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3721 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3781 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3841 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3901 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3961 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4021 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     4081 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4141 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     4201 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     4261 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     4321 tacggttcct ggtcttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     4381 attctgtgga taaccgtatt accgcctttg agtgagctga taccagcgga taacaatttc
     4441 aca
//
