LOCUS       sequence-449276-        7065 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mamalian cell expression of intermediate filaments (vimentin) fused 
            to SNAP-tag2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7065)
  AUTHORS   Kuhn S, Nasufovic V, Wilhelm J, Kompa J, de Lange EMF, Lin YH, 
            Egoldt C, Fischer J, Lennoi A, Tarnawski M, Reinstein J, Vlijm R, 
            Hiblot J, Johnsson K
  TITLE     SNAP-tag2 for faster and brighter protein labeling.
  JOURNAL   Nat Chem Biol. 2025 Jul 3. doi: 10.1038/s41589-025-01942-z.
  PUBMED    40610720
REFERENCE   2  (bases 1 to 7065)
  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: "Nat Chem 
            Biol. 2025 Jul 3. doi: 10.1038/s41589-025-01942-z."
COMMENT     Sequence Label: pcDNA5/FRT/TO_Vim-SNAP-tag2
FEATURES             Location/Qualifiers
     source          1..7065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1963..1987)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    1997..2015
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2018..2036
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(2038..2241)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2067..2087)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2242..2621
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2793..2812
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2887..2991
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2992..3060,3061..3852)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIGAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3210..3229)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4023..4610
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4512..4531
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4764..4781
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4898..4919
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4934..4939,4940..4957,4958..4964)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4972..4988
                     /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)."
     primer_bind     4977..4999
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4996..5012
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4996..5012
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5061..5182
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5072..5091)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5126..5145
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5312..6334)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             complement(5312..6334)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     protein_bind    complement(6342..6389)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     6521..6540
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    6690..6897
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6886..6903
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
ORIGIN
        1 tcagaattgg caccggatta ccacgcagag cggttttcac cgccgcagtt gccgctgggt
       61 tgccagccag cgccgccagc tgggaatagg agatcacctc gccgaatttc actactttca
      121 gcagtttcca cagtacctga cgggtaaagg attcctgctg aaataccggg tgatgcaggg
      181 ccggtacagg gaactcttca atcgcctctg gctggtgaaa atatgcgttc agccatgcgg
      241 tagcctgcat cagcggctcc gggcccggag gaccctgctc gccctgacca ttgcccacga
      301 aatagatgcg gtgcaggccc tgctcgcaac cgctcagcag cagtttaccc agcgggctgt
      361 cataggtagt gcgtttcatt tcgcagtctt tatcggtggc gaccggtgga tccccttcaa
      421 ggtcatcgtg atgctgagaa gtttcgttga taacctgtcc atctctagtt tcaaccgtct
      481 taatcagaag tgtccttttt gagtgggtat caaccagagg gagtgaatcc agattagttt
      541 ccctcaggtt cagggaggaa aagtttggaa gaggcagaga aatcctgctc tcctcgcctt
      601 ccagcagctt cctgtaggtg gcaatctcaa tgtcaagggc catcttaaca ttgagcaggt
      661 cttggtattc acgaaggtga cgagccattt cctccttcat attctgaatc tcatcctgca
      721 ggcggccaat agtgtcttgg tagttagcag cttcaacggc aaagttctct tccatttcac
      781 gcatctggcg ttccagggac tcattggttc ctttaagggc atccacttca caggtgaggg
      841 actgcacctg tctccggtac tcagtggact cctgctttgc ctggcgcagg gcgtcattgt
      901 tccggttggc agcctcagag aggtcagcaa acttggattt gtaccattct tctgcctcct
      961 gcaggttctt ggcagccaca ctttcatatt gctgacgtac gtcacgcagg gcagccgtga
     1021 ggtcaggctt ggaaacatcc acatcgattt ggacatgctg ttcctgaatc tgagcctgca
     1081 gctcctggat ttcctcttcg tggagtttct tcaaaaaggc aatctcttct tgcaaagatt
     1141 ccactttgcg ttcaaggtca agacgtgcca gagacgcatt gtcaacatcc tgtctgaaag
     1201 attgcagggt gttttcggct tcctctctct gaagcatctc ctcctgcaat ttctcccgga
     1261 ggcgcatgat gtcctcggcc aggttgtcgc gctccacctc gacgcgggct ttgtcgttgg
     1321 ttagctggtc cacctgccgg cgcagctccc gcatctcctc ctcgtagagg tcccccaggc
     1381 gcgacttgcc ttggcccttg agctgctcga gctcggccag caggatctta ttctgctgct
     1441 ccaggaagcg caccttgtcg atgtagttgg cgaagcggtc attcagctcc tgcagctcca
     1501 ccttctcgtt ggtgcgggtg ttcttgaact cggtgttgat ggcgtcggcc agcgagaagt
     1561 ccaccgagtc ctgcaggagc cgcaccccgg gcacgctgct ccgcaggcgc acggcagagg
     1621 agcgcgtggc atacacgccg cccggggacg aggcgtagag gctgcggctg gtgctggggc
     1681 gcagcgcgct gcccaggctg taggtgcggg tggacgtagt cacgtagctc cggctggagc
     1741 tcggccggct cgcggtgccc gggccgccga acatcctgcg gtaggaggac gaggacacgg
     1801 acctggtgga catggttgct ggatatctgc agaattccac cacactggac tagtggatcc
     1861 gagctcggta ccaagcttaa gtttaaacgc tagagtccgg aggctggatc ggtcccggtg
     1921 tcttctatgg aggtcaaaac agcgtggatg gcgtctccag gcgatctgac ggttcactaa
     1981 acgagctcgt cgacgatctc tatcactgat agggagatct ctatcactga tagggagagc
     2041 tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca atggggcgga
     2101 gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac
     2161 gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat
     2221 gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga tgtactgcca
     2281 agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc
     2341 attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc
     2401 agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat
     2461 gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg
     2521 gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc
     2581 cgtaattgat tactattaat aactagtcaa taatcaatgt caacgcgtat atctggcccg
     2641 tacatcgcga agcagcgcaa aacgcctaac cctaagcaga ttcttcatgc aattgtcggt
     2701 caagccttgc cttgttgtag cttaaatttt gctcgcgcac tactcagcga cctccaacac
     2761 acaagcaggg agcagatact ggcttaacta tgcggcatca gagcagattg tactgagagt
     2821 gcaccatagg ggatcgggag atctcccgat ccgtcgacgt caggtggcac ttttcgggga
     2881 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2941 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3001 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3061 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3121 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3181 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattggc
     3241 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3301 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3361 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3421 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3481 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3541 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3601 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3661 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3721 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3781 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3841 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     3901 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     3961 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4021 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4081 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4141 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4201 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4261 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4321 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4381 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4441 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4501 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4561 cttgagcgtc gatttttgtg atgctcgtca gggggcggag cctatggaaa aacgccagca
     4621 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     4681 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     4741 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     4801 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     4861 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     4921 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     4981 gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc tagctagagg
     5041 tcgacggtat acagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     5101 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     5161 tataagctgc aataaacaag ttggggtggg cgaagaactc cagcatgaga tccccgcgct
     5221 ggaggatcat ccagccggcg tcccggaaaa cgattccgaa gcccaacctt tcatagaagg
     5281 cggcggtgga atcgaaatct cgtagtacgt gctattcctt tgccctcgga cgagtgctgg
     5341 ggcgtcggtt tccactatcg gcgagtactt ctacacagcc atcggtccag acggccgcgc
     5401 ttctgcgggc gatttgtgta cgcccgacag tcccggctcc ggatcggacg attgcgtcgc
     5461 atcgaccctg cgcccaagct gcatcatcga aattgccgtc aaccaagctc tgatagagtt
     5521 ggtcaagacc aatgcggagc atatacgccc ggagccgcgg cgatcctgca agctccggat
     5581 gcctccgctc gaagtagcgc gtctgctgct ccatacaagc caaccacggc ctccagaaga
     5641 agatgttggc gacctcgtat tgggaatccc cgaacatcgc ctcgctccag tcaatgaccg
     5701 ctgttatgcg gccattgtcc gtcaggacat tgttggagcc gaaatccgcg tgcacgaggt
     5761 gccggacttc ggggcagtcc tcggcccaaa gcatcagctc atcgagagcc tgcgcgacgg
     5821 acgcactgac ggtgtcgtcc atcacagttt gccagtgata cacatgggga tcagcaatcg
     5881 cgcatatgaa atcacgccat gtagtgtatt gaccgattcc ttgcggtccg aatgggccga
     5941 acccgctcgt ctggctaaga tcggccgcag cgatcgcatc catggcctcc gcgaccggct
     6001 gcagaacagc gggcagttcg gtttcaggca ggtcttgcaa cgtgacaccc tgtgcacggc
     6061 gggagatgca ataggtcagg ctctcgctga attccccaat gtcaagcact tccggaatcg
     6121 ggagcgcggc cgatgcaaag tgccgataaa cataacgatc tttgtagaaa ccatcggcgc
     6181 agctatttac ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa gcacgagatt
     6241 cttcgccctc cgagagctgc atcaggtcgg agacgctgtc gaacttttcg atcagaaact
     6301 tctcgacaga cgtcgcggtg agttcaggct ttttggccaa ggaagttcct atactttcta
     6361 gagaatagga acttcggaat aggaacttct aggtacgtga accatcaccc taatcaagtt
     6421 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     6481 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     6541 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     6601 cgcttaatgc gccgctacag ggcgcgtggg gataccccct agagccccag ctggttcttt
     6661 ccgcctcaga agtcatagag cccaccgcat ccccagcatg cctgctattg tcttcccaat
     6721 cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac ctactcagac
     6781 aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca ccttccaggg
     6841 tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa ggcacagtcg
     6901 aggctgatca gcgggtttaa acgggccctc tagactcgag cggttagccc aggcctggtt
     6961 tacccagacg gtgaccttcg tgcgccagca gccattcttt cacgtacaga gggccttcgt
     7021 agccacctac gtcgccgtcg ccctgaacca cacggtggca cggga
//
