LOCUS       sequence-365684-        4651 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 4651)
  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 4651)
  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..4651
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(359..1150,1151..1219))
                     /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     982..1001
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1220..1324)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1350..1805)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1487..1508)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1699..1718
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1876..2860
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        2895..2913
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2895..2912
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             2948..3493
                     /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    3563..3697
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3587..3606)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3641..3660
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3810..3830)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3810..3828)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3849..3865)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3849..3865)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3862..3884)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3873..3889
                     /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(3897..3903,3904..3921,3922..3927))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3942..3963
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4080..4097)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(4251..4651,1..188))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4331..4350)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
       61 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
      121 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
      181 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
      241 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
      301 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
      361 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
      421 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
      481 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
      541 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
      601 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
      661 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
      721 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
      781 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
      841 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
      901 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
      961 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     1021 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     1081 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     1141 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     1201 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     1261 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     1321 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt
     1381 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     1441 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     1501 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     1561 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     1621 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     1681 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     1741 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     1801 cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     1861 ttcgctatta cgccagtcga ccgccaattc aatatggcgt atatggactc atgccaattc
     1921 aatatggtgg atctggacct gtgccaattc aatatggcgt atatggactc gtgccaattc
     1981 aatatggtgg atctggaccc cagccaattc aatatggcgg acttggcacc atgccaattc
     2041 aatatggcgg acctggcact gtgccaactg gggaggggtc tacttggcac ggtgccaagt
     2101 ttgaggaggg gtcttggccc tgtgccaagt ccgccatatt gaattggcat ggtgccaata
     2161 atggcggcca tattggctat atgccaggat caatatatag gcaatatcca atatggccct
     2221 atgccaatat ggctattggc caggttcaat actatgtatt ggccctatgc catatagtat
     2281 tccatatatg ggttttccta ttgacgtaga tagcccctcc caatgggcgg tcccatatac
     2341 catatatggg gcttcctaat accgcccata gccactcccc cattgacgtc aatggtctct
     2401 atatatggtc tttcctattg acgtcatatg ggcggtccta ttgacgtata tggcgcctcc
     2461 cccattgacg tcaattacgg taaatggccc gcctggctca atgcccattg acgtcaatag
     2521 gaccacccac cattgacgtc aatgggatgg ctcattgccc attcatatcc gttctcacgc
     2581 cccctattga cgtcaatgac ggtaaatggc ccacttggca gtacatcaat atctattaat
     2641 agtaacttgg caagtacatt actattggaa gtacgccagg gtacattggc agtactccca
     2701 ttgacgtcaa tggcggtaaa tggcccgcga tggctgccaa gtacatcccc attgacgtca
     2761 atggggaggg gcaatgacgc aaatgggcgt tccattgacg taaatgggcg gtaggcgtgc
     2821 ctaatgggag gtctatataa gcaatgctcg tttagggaac cgccattctg cctggggacg
     2881 tcggagcaag cttgatttag gtgacactat agaatacaag ctacttgttc tttttgcagg
     2941 atccaccatg gacaaagact gcgaaatgaa gcgcaccacc ctggatagcc ctctgggcaa
     3001 gctggaactg tctgggtgcg aacagggcct gcaccgtatc atcttcctgg gcaaaggaac
     3061 atctgccgcc gacgccgtgg aagtgcctgc cccagccgcc gtgctgggcg gaccagagcc
     3121 actgatgcag gccaccgcct ggctcaacgc ctactttcac cagcctgagg ccatcgagga
     3181 gttccctgtg ccagccctgc accacccagt gttccagcag gagagcttta cccgccaggt
     3241 gctgtggaaa ctgctgaaag tggtgaagtt cggagaggtc atcagctaca gccacctggc
     3301 cgccctggcc ggcaatcccg ccgccaccgc cgccgtgaaa accgccctga gcggaaatcc
     3361 cgtgcccatt ctgatcccct gccacagagt ggtgcagggc gacctggacg tggggggcta
     3421 cgagggcggg ctcgccgtga aagagtggct gctggcccac gagggccaca gactgggcaa
     3481 gcctgggctg ggttccggac tcagatctcg agctcaagct tcgaattctg cagtctagaa
     3541 ctatagtgag tcgtattacg tagatccaga catgataaga tacattgatg agtttggaca
     3601 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     3661 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     3721 tatgtttcag gttcaggggg aggtgtggga ggttttttaa ttcgcggccg cggcgccaat
     3781 gcattgggcc cggtacccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
     3841 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     3901 atacgagccg ggagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     3961 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4021 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4081 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4141 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4201 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4261 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4321 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4381 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4441 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4501 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4561 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4621 agcagagcga ggtatgtagg cggtgctaca g
//
