LOCUS       sequence-288728-        4211 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DrUPF3a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4211)
  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 4211)
  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-DrUPF3a_F
FEATURES             Location/Qualifiers
     source          1..4211
                     /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(644..663)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(645..663)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(670..687)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(670..686)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(679..701)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      827..1255
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(914..933)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1124..1145
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             1599..2393
                     /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(1653..1672)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2263..2282
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(2411..2479,2480..3271)
                     /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(2629..2648)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3442..4030
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3931..3950
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4184..4201
                     /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 gccctttatt ttgagttctt tcctgctgat
      361 cagagcttat accctcatct cttctcaaga gcatacataa acttcaaaaa cccagaggat
      421 atcattatat tcagagaccg ctttgatggc tatgtattta ttgacaataa aggtcaagaa
      481 tatccggctg ttgtagaatt tgccccattt cagaaggttt ccaagaaaaa gctaaagaaa
      541 aaagatgcga aagctgggac cattgaagaa ggcacagggc gaattctgca gatatccatc
      601 acactggcgg ccgctcgagc atgcatctag agggcccaat tcgccctata gtgagtcgta
      661 ttacaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
      721 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
      781 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggacgc gccctgtagc
      841 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
      901 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
      961 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     1021 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     1081 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     1141 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     1201 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     1261 aaaattcagg gcgcaagggc tgctaaagga agcggaacac gtagaaagcc agtccgcaga
     1321 aacggtgctg accccggatg aatgtcagct actgggctat ctggacaagg gaaaacgcaa
     1381 gcgcaaagag aaagcaggta gcttgcagtg ggcttacatg gcgatagcta gactgggcgg
     1441 ttttatggac agcaagcgaa ccggaattgc cagctggggc gccctctggt aaggttggga
     1501 agccctgcaa agtaaactgg atggctttct tgccgccaag gatctgatgg cgcaggggat
     1561 caagatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     1621 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     1681 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     1741 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     1801 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     1861 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tcccaccttg
     1921 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     1981 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     2041 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     2101 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     2161 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     2221 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     2281 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     2341 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgaattgaaa
     2401 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     2461 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     2521 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     2581 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     2641 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     2701 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     2761 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     2821 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     2881 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     2941 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3001 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     3061 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3121 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3181 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3241 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     3301 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3361 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3421 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3481 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3541 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3601 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3661 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3721 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3781 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3841 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3901 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3961 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4021 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4081 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     4141 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     4201 ggaagcggaa g
//
