LOCUS       sequence-288724-        4894 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DrSmg6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4894)
  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 4894)
  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-DrSmg6_F
FEATURES             Location/Qualifiers
     source          1..4894
                     /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(1327..1346)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1328..1346)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1353..1370)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1353..1369)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1362..1384)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1510..1938
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1597..1616)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1807..1828
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             2282..3076
                     /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(2336..2355)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2946..2965
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3094..3162,3163..3954)
                     /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(3312..3331)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4125..4713
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4614..4633
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4867..4884
                     /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 gccctttatg tcatagcaga ggcgggatac
      361 atcccattag agcctggtat ttggtgggga atctgatacg ggtatggata tcttggcggg
      421 gtgtttgggg aattggctga cataggatag cagtagggat tgtccgaatt ctggaacttg
      481 tagtacgcca tagctgcggc ctgctgattt tgaaagaact ccccctggcg gactggagga
      541 cttcctacgg tttcgtcatc agtgtcaaga aagtgtagcg gcccacatcc accctgctgt
      601 aaatacaaag cctggtgttg agatgcatgc tgggattgtt ggcttgacac cagtgctggt
      661 ttcttatctg ggttatttgg gtcccacaga cgtctggtcc ctcctccacg gccgcctctt
      721 cctctgccca ggcccattcc tcctctcatt cctccaagtg gaggtgcagg atcaggggtc
      781 tgtgttaggt cggtgtgggc aggaagcacc aggatgcctc ttccacgacc tctagtgctt
      841 ttacggcttg cttgctcctc acgcttgtct aaagacactc ttaagatccc tccttccttc
      901 cgagtgctgc tcttggcttc ttcagcagct ctgcttcttc tgccttctcg ctcccacacc
      961 tcgttttcct ccagcgagtc agtggaagac aaggtacggg ccgtgcgtct cctggagcgg
     1021 gtttgacctc ctcggacgaa ctccctctgt cccgtggctc tctctaaagt gcgtgcagaa
     1081 aactgctgac cttctgcttt caatctctcg gcttcagcaa gaccatccat actggtcccg
     1141 ctactggccg aactggtgct gtaggtgcgg tttcgaggcc gccgcatgtc tgattgagag
     1201 tatcgctttg aagcagagtt aactcgcttt gaatcagttc gctcagatgg ttttccttag
     1261 ggcgaattct gcagatatcc atcacactgg cggccgctcg agcatgcatc tagagggccc
     1321 aattcgccct atagtgagtc gtattacaat tcactggccg tcgttttaca acgtcgtgac
     1381 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     1441 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     1501 ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     1561 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     1621 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     1681 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     1741 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     1801 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     1861 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     1921 aaaaatttaa cgcgaatttt aacaaaattc agggcgcaag ggctgctaaa ggaagcggaa
     1981 cacgtagaaa gccagtccgc agaaacggtg ctgaccccgg atgaatgtca gctactgggc
     2041 tatctggaca agggaaaacg caagcgcaaa gagaaagcag gtagcttgca gtgggcttac
     2101 atggcgatag ctagactggg cggttttatg gacagcaagc gaaccggaat tgccagctgg
     2161 ggcgccctct ggtaaggttg ggaagccctg caaagtaaac tggatggctt tcttgccgcc
     2221 aaggatctga tggcgcaggg gatcaagatc tgatcaagag acaggatgag gatcgtttcg
     2281 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     2341 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     2401 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     2461 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     2521 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     2581 ggatctcctg tcatcccacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     2641 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     2701 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     2761 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     2821 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     2881 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     2941 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     3001 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     3061 tgacgagttc ttctgaattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     3121 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     3181 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     3241 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     3301 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     3361 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     3421 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     3481 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     3541 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     3601 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     3661 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     3721 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     3781 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     3841 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     3901 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     3961 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     4021 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4081 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4141 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4201 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4261 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4321 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4381 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4441 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4501 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4561 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4621 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4681 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     4741 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     4801 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     4861 gagcgcagcg agtcagtgag cgaggaagcg gaag
//
