LOCUS       sequence-288722-        5099 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DrSmg1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5099)
  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 5099)
  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-DrSmg1_F
FEATURES             Location/Qualifiers
     source          1..5099
                     /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(1532..1551)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1533..1551)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1558..1575)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1558..1574)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1567..1589)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1715..2143
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1802..1821)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2012..2033
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             2487..3281
                     /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(2541..2560)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3151..3170
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3299..3367,3368..4159)
                     /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(3517..3536)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4330..4918
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4819..4838
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5072..5089
                     /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 gccctttaca ggcatcaggc aaacccagag
      361 ggctcagcaa actgttcagt gcacagccca tctcgcccaa gaccagttta tatgccgcct
      421 ccaagatagg gatgttcttc aggcttagca cagcatgata cactccatgt gctgccgcca
      481 tcacctctct ctctgtgaaa gcgcagctcc aaaagatgag attttggggc caacaacttt
      541 tccacaaaag aagccggaag tttggtgttt atttggtcaa caaccagtgt gagcagggtg
      601 agaacagaca ggctgtattc gggtccgcag ttctggcagc agttcagctg gtctaggccg
      661 tatgagatta cagcatcaat cagctggccg cccaagtcta tgctcaccag cagaacacac
      721 acacactcat ttcctgctgt cagcaccgcc tcagagaaga acacctgctt ggctgtggtg
      781 acgcaagcca aaacacgatt caaaacatca gtgacatatg cttctgtaat tggagggcct
      841 cggattgggt taaaccgttc tccgatgctg tgcacaacag tgctaaaaac tctcaatagg
      901 gctgccaatt taggcaagga cacagtggga gggggaatgt cctcatctcc tgattctcca
      961 gatacaacat ggctcaggtc ctcagcatat gcctccatat cttccaaaaa ctgacccaat
     1021 aatgtggtcg agaaggccaa atctgcaacc caaaattgct ccaaactctg caaccagcca
     1081 gatacttgtt gagtaagaga gtgcttctgt gtgtgatcta tatgccagcc taccaagata
     1141 tccactgtgt ctctgaagtt ggtgctgaag atgtgtggat aacaacgggc caccagtaaa
     1201 atacacttta cactttggca caatagttcc ggtgtgtcca agttctccaa tatagactga
     1261 aggttgctca tcaccagctg cattacagct gaaaaggcct tcttctcccc agctgtttca
     1321 agtgctttgt gcactgccac aaggtacaga agcttcacct catcctttgt agacacactg
     1381 aacttgagga acatccactt gaagatcctc tcagcttcat aactgagggc ggcacagagc
     1441 aatcccagac accaagcggc ctcagggcga attctgcaga tatccatcac actggcggcc
     1501 gctcgagcat gcatctagag ggcccaattc gccctatagt gagtcgtatt acaattcact
     1561 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     1621 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     1681 ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg cgcattaagc
     1741 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     1801 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     1861 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     1921 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     1981 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2041 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2101 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aattcagggc
     2161 gcaagggctg ctaaaggaag cggaacacgt agaaagccag tccgcagaaa cggtgctgac
     2221 cccggatgaa tgtcagctac tgggctatct ggacaaggga aaacgcaagc gcaaagagaa
     2281 agcaggtagc ttgcagtggg cttacatggc gatagctaga ctgggcggtt ttatggacag
     2341 caagcgaacc ggaattgcca gctggggcgc cctctggtaa ggttgggaag ccctgcaaag
     2401 taaactggat ggctttcttg ccgccaagga tctgatggcg caggggatca agatctgatc
     2461 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     2521 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     2581 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     2641 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     2701 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     2761 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc ccaccttgct cctgccgaga
     2821 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     2881 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     2941 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3001 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     3061 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     3121 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     3181 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     3241 agcgcatcgc cttctatcgc cttcttgacg agttcttctg aattgaaaaa ggaagagtat
     3301 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3361 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     3421 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3481 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     3541 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     3601 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     3661 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     3721 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     3781 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     3841 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     3901 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     3961 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     4021 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     4081 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4141 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4201 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4261 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4321 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4381 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4441 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4501 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4561 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4621 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4681 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4741 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4801 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4861 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4921 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     4981 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     5041 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaag
//
