LOCUS       sequence-491020-        7303 bp    DNA     circular SYN 29-OCT-2025
DEFINITION  anchoring mCherry-1x SunTag to the intermediate filaments.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7303)
  AUTHORS   Vigano MA, Ell CM, Kustermann MMM, Aguilar G, Matsuda S, Zhao N, 
            Stasevich TJ, Affolter M, Pyrowolakis G
  TITLE     Protein manipulation using single copies of short peptide tags in 
            cultured cells and in Drosophila melanogaster.
  JOURNAL   Development. 2021 Mar 16;148(6):dev191700. doi: 10.1242/dev.191700.
  PUBMED    33593816
REFERENCE   2  (bases 1 to 7303)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 29, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Development."; date: "2021-03-16"; volume: "148(6):dev191700. doi";
            pages: " 10.1242/dev.191700"
COMMENT     Sequence Label: Vimentin_mCherry_SunTag
FEATURES             Location/Qualifiers
     source          1..7303
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2409..3113
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2551..2569)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2819..2838
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3186..3242
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     primer_bind     complement(3311..3328)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3317..3541
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3587..4015
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3674..3693)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3884..3905
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4024..4044)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4029..4358
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4209..4344
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4406..4453
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4472..4846
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4976..5097
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5013..5032)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5067..5086
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5146..5162)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5146..5162)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5159..5181)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5170..5186
                     /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(5194..5200,5201..5218,5219..5224))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5239..5260
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5377..5394)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5548..6136)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5628..5647)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6307..7098,7099..7167))
                     /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     6930..6949
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7168..7272)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtaccg agctcggatc catgtctacc aggtctgtgt
     1021 cctcgtcctc ctaccgcagg atgttcggtg gctccggcac atcgagccgg cccagctcca
     1081 accggagcta tgtgaccacg tccacacgca cctacagtct gggcagcgca ctgcgcccca
     1141 gcactagccg cagcctctat tcctcatccc ccggtggcgc ctatgtgacc cggtcctcgg
     1201 cagtgcgcct gcggagcagc gtgccgggcg tgcggctgct tcaagactcg gtggacttct
     1261 cgctggccga cgccatcaac actgagttca agaacacccg caccaacgag aaggtagaac
     1321 tgcaggagct gaatgaccgc tttgccaact acatcgacaa ggtgcgcttc ctcgagcagc
     1381 agaacaaaat cctgctggct gagctcgagc agctcaaggg ccagggcaag tcgcgcctgg
     1441 gcgacctgta cgaggaggag atgcgggagc tgcgccggca ggtggatcag ctcaccaacg
     1501 acaaggcccg tgtcgaggtg gagcgggaca acctggccga ggacatcatg cggctgcgag
     1561 agaaattgca ggaggagatg ctccagagag aggaagccga aagcaccctg cagtcattca
     1621 gacaggatgt tgacaatgct tctctggcac gtcttgacct tgaacggaaa gtggaatcct
     1681 tgcaggaaga aattgccttt ttgaagaaac tgcacgatga agagatccag gagctgcagg
     1741 cccagattca ggaacagcat gtccagatcg atgtggacgt ttccaagcct gacctcactg
     1801 ctgccctgcg tgatgtgcgc cagcagtatg aaagcgtggc tgccaagaac ctccaggagg
     1861 ccgaggaatg gtacaagtcc aagtttgctg acctctctga ggctgccaac cggaacaacg
     1921 atgccctgcg ccaggccaag caggagtcaa acgagtaccg gagacaggtg cagtcactca
     1981 cctgtgaagt ggatgccctt aaaggcacta acgagtccct ggagcgccag atgcgtgaga
     2041 tggaagagaa ttttgccctt gaagctgcta actaccagga cactattggc cgcctgcagg
     2101 atgagatcca aaacatgaag gaagagatgg ctcgtcacct tcgtgaatac caagatctgc
     2161 tcaatgttaa gatggccctg gacattgaga tcgccaccta caggaagctg ctggaaggcg
     2221 aggagagcag gatttctctg cctctgccaa ccttttcttc cctgaacctg agagaaacta
     2281 acctggagtc acttcctctg gttgacaccc actcaaaaag aacactcctg attaagacgg
     2341 ttgagaccag agatggacag gtgatcaatg agacttctca gcatcacgat gaccttgaag
     2401 aattcggtgt gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct
     2461 tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg
     2521 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc
     2581 tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga
     2641 agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg
     2701 agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc
     2761 aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc
     2821 ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg
     2881 acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg
     2941 acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca
     3001 acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     3061 acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac aagggtggtg
     3121 gtggcgcggc cgcagatcga tcgatcacgc gtgccaccat gactagtttt gtacaaagcg
     3181 gtccggaaga acttttgagc aagaattatc atcttgagaa cgaagtggct cgtcttaaga
     3241 aaaccgcggt aaacatatgg atcgatcata atactcgagt ctagagggcc cgtttaaacc
     3301 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3361 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3421 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3481 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     3541 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     3601 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3661 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3721 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3781 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3841 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3901 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3961 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     4021 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4081 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4141 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4201 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4261 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4321 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     4381 ccattttcgg atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat
     4441 agtataatac gacaaggtga ggaactaaac catggccaag ttgaccagtg ccgttccggt
     4501 gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc
     4561 ccgggacttc gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat
     4621 cagcgcggtc caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg
     4681 cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc
     4741 cgggccggcc atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga
     4801 cccggccggc aactgcgtgc acttcgtggc cgaggagcag gactgacacg tgctacgaga
     4861 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4921 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     4981 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5041 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5101 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     5161 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5221 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5281 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5341 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5401 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5461 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5521 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5581 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5641 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5701 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5761 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5821 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5881 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5941 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     6001 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6061 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6121 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6181 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6241 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6301 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6361 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6421 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6481 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6541 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6601 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6661 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6721 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6781 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6841 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6901 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6961 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7021 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7081 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7141 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7201 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7261 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
