LOCUS       sequence-362158-        4631 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  vector backbone to clone insert upstream of SNAPf tag (SNAPf tag 
            codon optimized for Xenopus), for mRNA synthesis .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4631)
  AUTHORS   Dudley CE, van den Goor L, Miller AL
  TITLE     SNAP- and Halo-tagging and dye introduction protocol for live 
            microscopy in Xenopus embryos.
  JOURNAL   STAR Protoc. 2022 Aug 18;3(3):101622. doi: 
            10.1016/j.xpro.2022.101622. eCollection 2022 Sep 16.
  PUBMED    36035797
REFERENCE   2  (bases 1 to 4631)
  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: "STAR 
            Protoc."; date: "2022-08-18"; volume: "3(3):101622. doi"; pages: " 
            10.1016/j.xpro.2022.101622. eCollection 2022 Sep 16"
COMMENT     Sequence Label: pCS2+/C-SNAPf
FEATURES             Location/Qualifiers
     source          1..4631
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        53..71
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     53..70
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             139..681
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="DKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVEV
                     PAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLKV
                     VKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLAV
                     KEWLLAHEGHRLGKPGLG"
     polyA_signal    701..835
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(725..744)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     779..798
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(948..968)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(948..966)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(987..1003)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(987..1003)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1000..1022)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1011..1027
                     /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)."
     promoter        complement(join(1035..1041,1042..1059,1060..1065))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1080..1101
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1218..1235)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1389..1977)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1469..1488)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2148..2939,2940..3008))
                     /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     2771..2790
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3009..3113)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3139..3594)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3276..3297)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3488..3507
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(3665..4631,1..18)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
       61 gacactatag aatacaagct acttgttctt tttgcaggat cccatcgatt cgaattcaag
      121 gcctctcgag cctctagaga caaagactgc gaaatgaagc gcaccaccct ggatagccct
      181 ctgggcaagc tggaactgtc tgggtgcgaa cagggcctgc accgtatcat cttcctgggc
      241 aaaggaacat ctgccgccga cgccgtggaa gtgcctgccc cagccgccgt gctgggcgga
      301 ccagagccac tgatgcaggc caccgcctgg ctcaacgcct actttcacca gcctgaggcc
      361 atcgaggagt tccctgtgcc agccctgcac cacccagtgt tccagcagga gagctttacc
      421 cgccaggtgc tgtggaaact gctgaaagtg gtgaagttcg gagaggtcat cagctacagc
      481 cacctggccg ccctggccgg caatcccgcc gccaccgccg ccgtgaaaac cgccctgagc
      541 ggaaatcccg tgcccattct gatcccctgc cacagagtgg tgcagggcga cctggacgtg
      601 gggggctacg agggcgggct cgccgtgaaa gagtggctgc tggcccacga gggccacaga
      661 ctgggcaagc ctgggctggg ttaatgtaca aggttacgta gatccagaca tgataagata
      721 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
      781 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
      841 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaatt
      901 cgcggccgcg gcgccaatgc attgggcccg gtacccagct tttgttccct ttagtgaggg
      961 ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     1021 ctcacaattc cacacaacat acgagccggg agcataaagt gtaaagcctg gggtgcctaa
     1081 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     1141 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     1201 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     1261 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     1321 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     1381 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     1441 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     1501 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     1561 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     1621 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     1681 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     1741 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     1801 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     1861 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     1921 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     1981 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     2041 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     2101 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     2161 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     2221 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     2281 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     2341 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     2401 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     2461 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     2521 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     2581 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     2641 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     2701 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     2761 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     2821 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     2881 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     2941 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3001 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     3061 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     3121 ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     3181 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     3241 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     3301 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     3361 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     3421 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     3481 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     3541 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt
     3601 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     3661 gccagtcgac cgccaattca atatggcgta tatggactca tgccaattca atatggtgga
     3721 tctggacctg tgccaattca atatggcgta tatggactcg tgccaattca atatggtgga
     3781 tctggacccc agccaattca atatggcgga cttggcacca tgccaattca atatggcgga
     3841 cctggcactg tgccaactgg ggaggggtct acttggcacg gtgccaagtt tgaggagggg
     3901 tcttggccct gtgccaagtc cgccatattg aattggcatg gtgccaataa tggcggccat
     3961 attggctata tgccaggatc aatatatagg caatatccaa tatggcccta tgccaatatg
     4021 gctattggcc aggttcaata ctatgtattg gccctatgcc atatagtatt ccatatatgg
     4081 gttttcctat tgacgtagat agcccctccc aatgggcggt cccatatacc atatatgggg
     4141 cttcctaata ccgcccatag ccactccccc attgacgtca atggtctcta tatatggtct
     4201 ttcctattga cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc ccattgacgt
     4261 caattacggt aaatggcccg cctggctcaa tgcccattga cgtcaatagg accacccacc
     4321 attgacgtca atgggatggc tcattgccca ttcatatccg ttctcacgcc ccctattgac
     4381 gtcaatgacg gtaaatggcc cacttggcag tacatcaata tctattaata gtaacttggc
     4441 aagtacatta ctattggaag tacgccaggg tacattggca gtactcccat tgacgtcaat
     4501 ggcggtaaat ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa tggggagggg
     4561 caatgacgca aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc taatgggagg
     4621 tctatataag c
//
