LOCUS       sequence-377311-        4357 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector containing MS2 coat protein fused to ADAR2 
            catalytic domain (E488Q/T490A) with V5 tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4357)
  TITLE     Zebrafish orthologs of chromosome 21 genes
REFERENCE   2  (bases 1 to 4357)
  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"
COMMENT     Sequence Label: pTwist-ENTR-MCP-ADAR2-DD(E488Q/T490A)-V5
FEATURES             Location/Qualifiers
     source          1..4357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             7..27
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             333..344
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1942..1983
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    complement(1988..2087)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2144..2160)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2144..2160)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2157..2179)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(2332..2920)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2412..2431)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3056..3871)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3056..3871)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3779..3798
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3863..3882
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     3992..4008
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3992..4008
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      4009..4055
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4169..4191
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4183..4200
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4184..4200
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4253..4352
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 atgggaccta aaaaaaaacg aaaggtgggc gtcaagatgg cttccaattt cactcagttt
       61 gttctggtgg ataatggggg caccggtgac gtaacagtag caccgagtaa ctttgccaac
      121 ggaatagctg aatggatctc aagtaacagc cgtagtcagg catacaaggt gacttgtagc
      181 gttaggcaga gtagtgcaca aaataggaag tacaccatca aagtggaggt acccaaaggt
      241 gcttggagat cttatctcaa tatggagctc actataccca tattcgctac caactcagat
      301 tgcgagctga ttgtaaaggc aatgcagggc ttattgaagg acggaaatcc tatcccatct
      361 gctatcgctg ccaacagtgg aatttacgct atggccagca atttcaccca gttcgtcctg
      421 gttgataacg gtggcaccgg cgacgttact gtggctccaa gtaactttgc caacggcatt
      481 gcagagtgga taagctccaa tagtcgtagt caagcttata aagttacctg cagtgttagg
      541 cagagtagcg cccagaacag aaagtatacg attaaagtag aggttcccaa gggcgcttgg
      601 aggtcctatt taaacatgga actcactatc cctatatttg ctactaactc tgactgcgaa
      661 ctgatagtga aagccatgca gggattgctg aaggatggta accctatacc ttcagccatt
      721 gctgcaaatt ccgggggcgg ttctggcgga acaggtggat ccggcggtac tcaactgcat
      781 ctgccccagg ttttggccga tgccgtttca aggttggttt taggcaaatt tggagatctg
      841 actgacaact tctcttctcc tcatgctagg aggaaagttc tcgctggcgt tgtcatgact
      901 acaggaaccg acgtaaagga tgctaaagtc atctcagttt caaccggcac aaaatgcata
      961 aatggggaat atatgtcaga ccggggactt gccctcaacg actgccatgc agagatcata
     1021 tctcgccggt ccctgctgag atttttgtac acccaactcg aactgtacct gaataataag
     1081 gacgaccaaa agaggagcat tttccagaag tcagagagag gggggtttag actgaaggag
     1141 aatgttcaat tccacctgta catctccacg tctccttgcg gcgacgcacg catatttagt
     1201 ccccacgaac ctatacttga ggagcctgcc gatagacacc caaatcgtaa ggctagaggt
     1261 caactacgca ctaaaattga gtccgggcaa ggtgccatac ccgtaaggag caatgccagc
     1321 atccagacct gggacggggt tcttcaagga gaacgacttc tcaccatgag ttgtagcgac
     1381 aaaatagctc gctggaacgt tgttgggatc caaggatcac tgctatccat cttcgtagag
     1441 ccaatatatt tctcatcaat cattctaggc agtctgtatc acggtgacca tttgagccgc
     1501 gcaatgtacc aaaggatttc aaacattgaa gatctgcctc cactgtacac tttgaacaag
     1561 cccctcctga gtggcatatc caacgctgaa gcaaggcaac caggcaaagc accaaacttt
     1621 agcgttaact ggacagtggg ggatagcgcc atagaagtta tcaatgccac aaccggtaag
     1681 gatgaactag gaagagcaag cagactgtgc aagcacgcac tatactgccg atggatgcgc
     1741 gtccacggaa aggttccatc tcacctgttg agatcaaaaa ttactaagcc taatgtatac
     1801 cacgaaagta aattagccgc taaagaatac caggccgcta aagcacggct gtttactgca
     1861 tttataaaag ccggccttgg agcctgggtt gaaaaaccca ctgagcagga ccagttcagc
     1921 ttgaccggcg gcggaggcag tggcaaaccg attccaaacc ctttgttggg actagactct
     1981 acataagacc cagctttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt
     2041 tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgagg ctaggtggag
     2101 gctcagtgat gataagtctg cgatggtgga tgcatgtgtc atggtcatag ctgtttcctg
     2161 tgtgaaattg ttatccgctc agagggcaca atcctattcc gcgctatccg acaatctcca
     2221 agacattagg tggagttcag ttcggcgtat ggcatatgtc gctggaaaga acatgtgagc
     2281 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2341 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2401 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2461 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2521 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2581 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2641 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2701 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2761 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2821 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2881 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2941 tacggggtct gacgctctat tcaacaaagc cgccgtcccg tcaagtcagc gtaaatgggt
     3001 agggggcttc aaatcgtccg ctctgccagt gttacaacca attaacaaat tctgattaga
     3061 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
     3121 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
     3181 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     3241 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
     3301 ccggtgagaa tggcaaaagc ttatgcattt ctttccagac ttgttcaaca ggccagccat
     3361 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
     3421 gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
     3481 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
     3541 ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag
     3601 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     3661 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     3721 ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat
     3781 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctcg
     3841 agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag
     3901 cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat
     3961 tttgagacac aacgtggctt tcccccgccg ctctagaact agtggatcca aataaaacga
     4021 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcgca ttatacgaga
     4081 cgtccaggtt gggatacctg aaacaaaacc catcgtacgg ccaaggaagt ctccaataac
     4141 tgtgatccac cacaagcgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     4201 catgcataat ccgcacgcat ctggaataag gaagtgccat tccgcctgac ctcaaataat
     4261 gattttattt tgactgatag tgacctgttc gttgcaacag attgatgagc aatgcttttt
     4321 tataatgcca actttgtaca aaaaagcagg ctgcacc
//
