LOCUS       sequence-357418-        4778 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  vector backbone to clone insert downstream 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 4778)
  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 4778)
  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+/N-SNAPf
FEATURES             Location/Qualifiers
     source          1..4778
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             88..633
                     /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="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     polyA_signal    703..837
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(727..746)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     781..800
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(950..970)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(950..968)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(989..1005)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(989..1005)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1002..1024)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1013..1029
                     /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(1037..1043,1044..1061,1062..1067))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1082..1103
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1220..1237)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1391..1979)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1471..1490)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2150..2941,2942..3010))
                     /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     2773..2792
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3011..3115)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3141..3596)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3278..3299)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3490..3509
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3670..4654
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccaccatg gacaaagact gcgaaatgaa gcgcaccacc
      121 ctggatagcc ctctgggcaa gctggaactg tctgggtgcg aacagggcct gcaccgtatc
      181 atcttcctgg gcaaaggaac atctgccgcc gacgccgtgg aagtgcctgc cccagccgcc
      241 gtgctgggcg gaccagagcc actgatgcag gccaccgcct ggctcaacgc ctactttcac
      301 cagcctgagg ccatcgagga gttccctgtg ccagccctgc accacccagt gttccagcag
      361 gagagcttta cccgccaggt gctgtggaaa ctgctgaaag tggtgaagtt cggagaggtc
      421 atcagctaca gccacctggc cgccctggcc ggcaatcccg ccgccaccgc cgccgtgaaa
      481 accgccctga gcggaaatcc cgtgcccatt ctgatcccct gccacagagt ggtgcagggc
      541 gacctggacg tggggggcta cgagggcggg ctcgccgtga aagagtggct gctggcccac
      601 gagggccaca gactgggcaa gcctgggctg ggttccggac tcagatctcg agctcaagct
      661 tcgaattctg cagtctagaa ctatagtgag tcgtattacg tagatccaga catgataaga
      721 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
      781 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
      841 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
      901 ttcgcggccg cggcgccaat gcattgggcc cggtacccag cttttgttcc ctttagtgag
      961 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     1021 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     1081 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     1141 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     1201 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     1261 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     1321 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     1381 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     1441 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1501 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     1561 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     1621 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     1681 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     1741 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     1801 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     1861 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     1921 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     1981 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     2041 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     2101 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     2161 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     2221 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     2281 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     2341 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     2401 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     2461 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     2521 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     2581 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     2641 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     2701 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     2761 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     2821 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     2881 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     2941 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     3001 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     3061 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     3121 ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     3181 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     3241 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     3301 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     3361 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     3421 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     3481 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     3541 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca
     3601 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     3661 acgccagtcg accatagcca attcaatatg gcgtatatgg actcatgcca attcaatatg
     3721 gtggatctgg acctgtgcca attcaatatg gcgtatatgg actcgtgcca attcaatatg
     3781 gtggatctgg accccagcca attcaatatg gcggacttgg caccatgcca attcaatatg
     3841 gcggacttgg cactgtgcca actggggagg ggtctacttg gcacggtgcc aagtttgagg
     3901 aggggtcttg gccctgtgcc aagtccgcca tattgaattg gcatggtgcc aataatggcg
     3961 gccatattgg ctatatgcca ggatcaatat ataggcaata tccaatatgg ccctatgcca
     4021 atatggctat tggccaggtt caatactatg tattggccct atgccatata gtattccata
     4081 tatgggtttt cctattgacg tagatagccc ctcccaatgg gcggtcccat ataccatata
     4141 tggggcttcc taataccgcc catagccact cccccattga cgtcaatggt ctctatatat
     4201 ggtctttcct attgacgtca tatgggcggt cctattgacg tatatggcgc ctcccccatt
     4261 gacgtcaatt acggtaaatg gcccgcctgg ctcaatgccc attgacgtca ataggaccac
     4321 ccaccattga cgtcaatggg atggctcatt gcccattcat atccgttctc acgcccccta
     4381 ttgacgtcaa tgacggtaaa tggcccactt ggcagtacat caatatctat taatagtaac
     4441 ttggcaagta cattactatt ggaaggacgc cagggtacat tggcagtact cccattgacg
     4501 tcaatggcgg taaatggccc gcgatggctg ccaagtacat ccccattgac gtcaatgggg
     4561 aggggcaatg acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc gtgcctaatg
     4621 ggaggtctat ataagcaatg ctcgtttagg gaaccctggt nctactcact tctccttctc
     4681 tactnactaa ctctntctag cttgctacct nctctaacta ccctncttct cacctcctnc
     4741 taacagngct nactactcct nctcaatgta cctcaagt
//
