LOCUS       sequence-181934-        5691 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of SNAPf-Nup43.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5691)
  AUTHORS   Finan K, Raulf A, Heilemann M
  TITLE     A set of homo-oligomeric standards allows accurate protein counting.
  JOURNAL   Angew Chem Int Ed Engl. 2015 Oct 5;54(41):12049-52. doi: 
            10.1002/anie.201505664. Epub 2015 Aug 20.
  PUBMED    26289028
REFERENCE   2  (bases 1 to 5691)
  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"; doi: 
            "10.1002/anie.201505664"; journalName: "Angew Chem Int Ed Engl"; 
            date: "2015-10-5- 5"; volume: "54"; issue: "41"; pages: "12049-52"
COMMENT     Sequence Label: pSNAPf-Nup43
FEATURES             Location/Qualifiers
     source          1..5691
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      19..607
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     508..527
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        782..1085
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1086..1289
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1240..1260
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1322..1331
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1328..1873
                     /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    3200..3321
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3237..3256)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3291..3310
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3328..3783)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3465..3486)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3677..3696
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3810..3914
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3916..4273
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3916..4107
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3939..3959)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4124..4259
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4308..5102
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4308..5102
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4362..4381)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4972..4991
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5290..5309)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5334..5381
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
       61 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
      121 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
      181 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      241 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
      301 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      361 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      421 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
      481 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      541 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      601 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
      661 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
      721 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
      781 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      841 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      901 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      961 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     1021 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     1081 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     1141 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     1201 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     1261 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgccaccggt
     1321 cgccaccatg gacaaagact gcgaaatgaa gcgcaccacc ctggatagcc ctctgggcaa
     1381 gctggaactg tctgggtgcg aacagggcct gcaccgtatc atcttcctgg gcaaaggaac
     1441 atctgccgcc gacgccgtgg aagtgcctgc cccagccgcc gtgctgggcg gaccagagcc
     1501 actgatgcag gccaccgcct ggctcaacgc ctactttcac cagcctgagg ccatcgagga
     1561 gttccctgtg ccagccctgc accacccagt gttccagcag gagagcttta cccgccaggt
     1621 gctgtggaaa ctgctgaaag tggtgaagtt cggagaggtc atcagctaca gccacctggc
     1681 cgccctggcc ggcaatcccg ccgccaccgc cgccgtgaaa accgccctga gcggaaatcc
     1741 cgtgcccatt ctgatcccct gccaccgggt ggtgcagggc gacctggacg tggggggcta
     1801 cgagggcggg ctcgccgtga aagagtggct gctggcccac gagggccaca gactgggcaa
     1861 gcctgggctg ggtgctagcg gagcatccgc aggaatggag gaaatttatg cgaagtttgt
     1921 gtcccagaaa atcagcaaaa cccgctggcg accgctgcct ccgggaagtt tacagaccgc
     1981 ggagacgttc gctacaggat cttgggacaa tgaggaaaat tatatttcac tgtggtctat
     2041 tggagatttt ggaaacttgg actctgatgg agggtttgaa ggagaccatc agttattgtg
     2101 tgatatcaga caccatggtg atgtaatgga tttacagttt tttgaccagg aaagaattgt
     2161 cgctgcttca tcaacaggat gtgtaacagt tttccttcac catccaaata accagactct
     2221 gtcagtcaac cagcagtgga ctacagctca ctaccacaca ggccctggca gtccttccta
     2281 tagcagtgca ccatgtacag gtgttgtgtg caacaaccca gaaatcgtta cagttggaga
     2341 ggatggtcga ataaatctct tcagagctga tcacaaggaa gctgtaagaa ccatagacaa
     2401 tgcagatagt agtacactcc atgctgtaac ctttcttcga actcctgaga ttcttactgt
     2461 aaattcaatt ggacagttga aaatatggga tttcagacaa caaggaaatg agccttctca
     2521 gatattgtca ctgactggtg accgagtgcc actccactgt gttgatagac atcccaacca
     2581 acagcatgtt gtagctactg gtggccaaga tggaatgttg agtatttggg atgttagaca
     2641 aggtactatg cctgtatctc tgctgaaggc tcatgaagct gaaatgtggg aagttcactt
     2701 tcacccatcc aacccagaac atctttttac ctgctctgaa gatggatccc tctggcactg
     2761 ggatgcttcc acagatgtac ctgaaaagtc gtcactcttt caccaaggag gaagaagcag
     2821 tacttttttg tctcatagca ttagtaacca agctaatgtt caccagtctg tcattagctc
     2881 ctggctcagc actgatcctg caaaagaccg aattgaaatc acaagcttac ttcccagtag
     2941 gtctctgtct gtgaacactt tggatgtttt aggtccttgt cttgtttgtg gaaccgatgc
     3001 agaagcaatt tatgttacta gacatctttt ttcgtagcgg ccgcatcgat aacaccgcaa
     3061 atgcttctag ggatccaccg gatctagata actgatcata atcagccata ccacatttgt
     3121 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     3181 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     3241 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     3301 caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     3361 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     3421 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     3481 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     3541 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     3601 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     3661 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     3721 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     3781 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3841 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3901 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     3961 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4021 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4081 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     4141 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4201 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4261 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     4321 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4381 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4441 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     4501 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4561 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4621 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4681 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4741 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4801 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     4861 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4921 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4981 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     5041 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     5101 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5161 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5221 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     5281 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     5341 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     5401 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     5461 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     5521 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     5581 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5641 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag a
//
