LOCUS       sequence-288727-        5270 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DrUPF2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5270)
  AUTHORS   Wittkopp N, Huntzinger E, Weiler C, Sauliere J, Schmidt S, Sonawane 
            M, Izaurralde E
  TITLE     Nonsense-mediated mRNA decay effectors are essential for zebrafish 
            embryonic development and survival.
  JOURNAL   Mol Cell Biol. 2009 Jul . 29(13):3517-28.
  PUBMED    19414594
REFERENCE   2  (bases 1 to 5270)
  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. 2009 Jul . 29(13):3517-28."
COMMENT     Sequence Label: pCRII-TOPO-DrUPF2_F
FEATURES             Location/Qualifiers
     source          1..5270
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        239..257
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     239..256
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1703..1722)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1704..1722)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1729..1746)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1729..1745)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1738..1760)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1886..2314
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1973..1992)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2183..2204
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             2658..3452
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2712..2731)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3322..3341
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3470..3538,3539..4330)
                     /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(3688..3707)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4501..5089
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4990..5009
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5243..5260
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat
      241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcca
      301 ctagtaacgg ccgccagtgt gctggaattc gcccttctcc atgccgaggc gaatatcctc
      361 cagcactccg tccaccacat gaatgcccac gtcctcctga tacgccacca gtcctgccag
      421 caggtttgcc acacagtgga tgctgttata cttcacgttc cagatgttga ccatacagca
      481 gatcaggtaa cccttcacct cactgtcatg ccacggcagc ttacgcatct gcctnaaaac
      541 cttctcagtg gtgactttgg acaggtcttt atagagcagt tttcggatgt actcctgcag
      601 cggaggcctc ttcctgcgca ccgtcttctc catcggcgga gggttgcagt agtaataggc
      661 gttttccacc atggtcacgt agcgcacatc cagatgctga gcctgcttct tgcgcatcat
      721 ctgctccagc aggacactcg tgcgcaggtg agaatctgga gagcggaaga ggaaacgtcc
      781 acacgtctcc agcagcgtgc aggccatctc gatatgatga tgagagaaat cagacagcag
      841 catcttnnng ncagtgcagc gtgtcngttt tggagaacat cnttaaactt ggccaactct
      901 ccgatgaacc tcaccgtctt attcttggtc tcnatgttga tttggntcct tctttctgat
      961 atgaaatctg aagtctcctt tgagcatgga gcagaggtct tcagccacgt cagacataca
     1021 cgggntgaag agtggccacc agacgggcgt aaaacggcag cagatccaac ctctgtctgg
     1081 gcactgtgaa cagagcacgg actaacttcc tgcggtttga cttggtgttc atgttcatgc
     1141 agaagtccat tgcagccttg tcgatcaggt ctctgttaat gcagtttggc agctgctgaa
     1201 tgaaagcgtc cacgatgagc tttagatgag agccggtgct ggcctcctca tcctcctgct
     1261 cttgctcatc cagcagtttt ttggccagtt cttcactctc cacttcatcg gttgcatcca
     1321 gatccatcgg ttcatcagcg atgtccagcg cctccagctc catctccagc tcctcagtgc
     1381 tgtttccctc tcggctttct ttgctgctct ggctgctctt ctcattgtct ttgaacagga
     1441 ttgcaggaac aaacgctttc agatccacca tgttctcata gaagtttctt gcatcttcat
     1501 cctcccagat tcctccttct agatcatact ctcctggctt ccccggtgtg aagatgtcga
     1561 tgcctggacc gtgcttcctg tacggttttg tcctggggca gttcggcatg ttctcgtcca
     1621 gcaggtcggc cagaagggcg aattctgcag atatccatca cactggcggc cgctcgagca
     1681 tgcatctaga gggcccaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt
     1741 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     1801 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     1861 gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     1921 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     1981 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2041 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2101 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2161 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     2221 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     2281 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaattcaggg cgcaagggct
     2341 gctaaaggaa gcggaacacg tagaaagcca gtccgcagaa acggtgctga ccccggatga
     2401 atgtcagcta ctgggctatc tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag
     2461 cttgcagtgg gcttacatgg cgatagctag actgggcggt tttatggaca gcaagcgaac
     2521 cggaattgcc agctggggcg ccctctggta aggttgggaa gccctgcaaa gtaaactgga
     2581 tggctttctt gccgccaagg atctgatggc gcaggggatc aagatctgat caagagacag
     2641 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     2701 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     2761 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     2821 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     2881 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     2941 gcgaagtgcc ggggcaggat ctcctgtcat cccaccttgc tcctgccgag aaagtatcca
     3001 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     3061 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     3121 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     3181 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     3241 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     3301 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     3361 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     3421 ccttctatcg ccttcttgac gagttcttct gaattgaaaa aggaagagta tgagtattca
     3481 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3541 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     3601 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     3661 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     3721 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     3781 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     3841 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     3901 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     3961 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4021 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4081 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4141 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4201 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4261 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4321 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     4381 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4441 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4501 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4561 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4621 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     4681 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4741 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4801 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4861 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4921 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4981 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5041 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5101 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     5161 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     5221 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
//
