LOCUS       sequence-288723-        5121 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DrSmg5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5121)
  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 5121)
  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-DrSmg5_F
FEATURES             Location/Qualifiers
     source          1..5121
                     /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(1554..1573)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1555..1573)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1580..1597)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1580..1596)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1589..1611)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1737..2165
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1824..1843)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2034..2055
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             2509..3303
                     /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(2563..2582)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3173..3192
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3321..3389,3390..4181)
                     /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(3539..3558)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4352..4940
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4841..4860
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5094..5111
                     /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 gcccttcttt cctcctcctc ttcatcgtca
      361 tcctcctcca gagagccgtt ctgcagggtt cgctcctctg agagagcagc actactgtga
      421 tctctggcat cagcgtcatc agtgttatcg gtgcgcaggg ctgggacatc actctctccc
      481 tcctccaatt cagcttgcag tcgaatgttg acatgattga ccagatggga gaacagagcc
      541 aatgtgaaag cgatggaggc gctgtactgc tttgaagccc ctcgcttcag gctgtgcacc
      601 accatcagac aaatcaccac catcttgaaa ataaggctgt ctggaagcac agaaccaaca
      661 gagtcgtgct cttcctcctc ctcgctggcc gactggctgc catgagcagg aagaggcagg
      721 tagaaaagca ccaggttaaa gtcttccagc acagactggc acagcgacgt cagctccgtc
      781 tccatcagac tgttcttggg ctgtaacagg ctctgcaaat acatgaagct caccagcagt
      841 cttttgatgt cttttgacct ctgtcgagat ggtgaaagct tcttcatttc ttgtttcttc
      901 acctggtggt acatcttgga tgccttgtca aacagacgct ttaaattccc atacgcaccg
      961 tcgaagggag tctctgactg agtacagcgc aagtagtagt aagtagcctc cacattataa
     1021 aatttacttc cggcaagcgt acccaactga ttaaacggca tgccgacgtg aggtgcgaca
     1081 gacagggcct ggtgatagaa acgctccgcc agctgctcgg cctccactcc cgccagttca
     1141 ttctggtagc gcgcgagatc tcccaagtac acgaggcatc tgtgacacgc catctgggcc
     1201 cattccatct ccttagaggt ggctgagact ggtttcttac gaccaatgag cgggtcagtg
     1261 acatgtgtcc agtctatgca gtcctgtaac tccagctgat agtgtgactg gatgtagagc
     1321 agcaggtgtt gaaaaaaagc ccaccccggc aatcagatgc gtcctgtagg cacactccag
     1381 agcactgcgg ctgtgaatat gcttcttgtt ggtcttgatc acctgaatgg cctcatagta
     1441 gactttcctc cacagcagct cttctgcctt gcgtccataa tccacagggc gaattctgca
     1501 gatatccatc acactggcgg ccgctcgagc atgcatctag agggcccaat tcgccctata
     1561 gtgagtcgta ttacaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     1621 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     1681 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggacgc
     1741 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     1801 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     1861 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     1921 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     1981 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2041 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     2101 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2161 gaattttaac aaaattcagg gcgcaagggc tgctaaagga agcggaacac gtagaaagcc
     2221 agtccgcaga aacggtgctg accccggatg aatgtcagct actgggctat ctggacaagg
     2281 gaaaacgcaa gcgcaaagag aaagcaggta gcttgcagtg ggcttacatg gcgatagcta
     2341 gactgggcgg ttttatggac agcaagcgaa ccggaattgc cagctggggc gccctctggt
     2401 aaggttggga agccctgcaa agtaaactgg atggctttct tgccgccaag gatctgatgg
     2461 cgcaggggat caagatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     2521 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     2581 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     2641 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     2701 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     2761 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     2821 tcccaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     2881 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     2941 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     3001 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     3061 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     3121 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     3181 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     3241 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     3301 tgaattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     3361 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     3421 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     3481 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     3541 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     3601 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     3661 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     3721 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     3781 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     3841 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     3901 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     3961 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     4021 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     4081 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     4141 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     4201 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4261 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4321 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4381 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4441 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     4501 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4561 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4621 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4681 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4741 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4801 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4861 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4921 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4981 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     5041 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     5101 cagtgagcga ggaagcggaa g
//
