LOCUS       sequence-376484-        6024 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construct for labelling microtubules in mammalian cells with 
            mScarlet3-alphatubulin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6024)
  AUTHORS   Gadella TWJ Jr, van Weeren L, Stouthamer J, Hink MA, Wolters AHG, 
            Giepmans BNG, Aumonier S, Dupuy J, Royant A
  TITLE     mScarlet3: a brilliant and fast-maturing red fluorescent protein.
  JOURNAL   Nat Methods. 2023 Apr;20(4):541-545. doi: 
            10.1038/s41592-023-01809-y. Epub 2023 Mar 27.
  PUBMED    36973546
REFERENCE   2  (bases 1 to 6024)
  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: 
            "10.1038/s41592-023-01809-y"; journalName: "Nat Methods"; date: 
            "2023-04"; volume: "20"; issue: "4"; pages: "541-545"
COMMENT     Sequence Label: pmScarlet3-alphaTubulin_C1
FEATURES             Location/Qualifiers
     source          1..6024
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             169..963
                     /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             169..963
                     /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(223..242)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     833..852
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1151..1170)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1195..1242
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1571..2159
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2060..2079
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2334..2637
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2638..2841
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2792..2812
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2880..2889
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2886..3572
                     /codon_start=1
                     /product="bright cysteine-free monomeric red fluorescent 
                     protein with fast and complete maturation (Gadella et al., 
                     2023)"
                     /label=mScarlet3
                     /translation="MDSTEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LRVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYWKQSFPEGFKWERVMNFEDGGA
                     VSVAQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDVVLKGDIKM
                     ALRLKDGGRYLADFKTTYRAKKPVQMPGAFNIDRKLDITSHNEDYTVVEQYERSVARHS
                     TGGSGGS"
     primer_bind     3284..3303
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5085..5206
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5122..5141)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5176..5195
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5213..5668)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5350..5371)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5562..5581
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5695..5799
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(5801..6024,1..134)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5801..5992
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5824..5844)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(6009..6024,1..120)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
       61 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
      121 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
      181 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
      241 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
      301 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
      361 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
      421 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
      481 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
      541 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
      601 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
      661 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
      721 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
      781 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
      841 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
      901 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
      961 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     1021 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     1081 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     1141 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     1201 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     1261 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     1321 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     1381 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     1441 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     1501 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     1561 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     1621 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     1681 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     1741 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     1801 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     1861 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     1921 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     1981 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     2041 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     2101 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     2161 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     2221 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat
     2281 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     2341 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     2401 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     2461 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     2521 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     2581 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
     2641 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
     2701 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     2761 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
     2821 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg
     2881 ccaccatgga tagcaccgag gcagtgatca aggagttcat gcggttcaag gtgcacatgg
     2941 agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
     3001 agggcaccca gaccgccaag ctgagggtga ccaagggtgg ccccctgccc ttctcctggg
     3061 acatcctgtc ccctcagttc atgtacggct ccagggcctt cacgaagcac cccgccgaca
     3121 tccccgacta ctggaagcag tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     3181 tcgaggacgg cggcgccgtg tccgtggccc aggacacctc cctggaggac ggcaccctga
     3241 tctacaaggt gaagctccgc ggcaccaact tccctcctga cggccccgta atgcagaaga
     3301 agacaatggg ctgggaagca tccaccgagc ggttgtaccc cgaggacgtc gtgctgaagg
     3361 gcgacattaa gatggccctg cgcctgaagg acggcggccg ctacctggcg gacttcaaga
     3421 ccacctacag ggccaagaag cccgtgcaga tgcccggcgc cttcaacatc gaccgcaagt
     3481 tggacatcac atcccacaac gaggactaca ccgtggtgga acagtacgaa cgctccgtgg
     3541 cccgccactc caccggcggc tccggtggct ccttgtacaa gtccggactc agatctcgag
     3601 tgcgtgagtg catctccatc cacgttggcc aggctggtgt ccagattggc aatgcctgct
     3661 gggagctcta ctgcctggaa cacggcatcc agcccgatgg ccagatgcca agtgacaaga
     3721 ccattggggg aggagatgac tccttcaaca ccttcttcag tgagacgggc gctggcaagc
     3781 acgtgccccg ggctgtgttt gtagacttgg aacccacagt cattgatgaa gttcgcactg
     3841 gcacctaccg ccagctcttc caccctgagc agctcatcac aggcaaggaa gatgctgcca
     3901 ataactatgc ccgagggcac tacaccattg gcaaggagat cattgacctt gtgttggacc
     3961 gaattcgcaa gctggctgac cagtgcaccg gtcttcaggg cttcttggtt ttccacagct
     4021 ttggtggggg aactggttct gggttcacct ccctgctcat ggaacgtctc tcagttgatt
     4081 atggcaagaa gtccaagctg gagttctcca tttacccagc accccaggtt tccacagctg
     4141 tagttgagcc ctacaactcc atcctcacca cccacaccac cctggagcac tctgattgtg
     4201 ccttcatggt agacaatgag gccatctatg acatctgtcg tagaaacctc gatatcgagc
     4261 gcccaaccta cactaacctt aaccgcctta ttagccagat tgtgtcctcc atcactgctt
     4321 ccctgagatt tgatggagcc ctgaatgttg acctgacaga attccagacc aacctggtgc
     4381 cctacccccg catccacttc cctctggcca catatgcccc tgtcatctct gctgagaaag
     4441 cctaccatga acagctttct gtagcagaga tcaccaatgc ttgctttgag ccagccaacc
     4501 agatggtgaa atgtgaccct cgccatggta aatacatggc ttgctgcctg ttgtaccgtg
     4561 gtgacgtggt tcccaaagat gtcaatgctg ccattgccac catcaaaacc aagcgcagca
     4621 tccagtttgt ggattggtgc cccactggct tcaaggttgg catcaactac cagcctccca
     4681 ctgtggtgcc tggtggagac ctggccaagg tacagagagc tgtgtgcatg ctgagcaaca
     4741 ccacagccat tgctgaggcc tgggctcgcc tggaccacaa gtttgacctg atgtatgcca
     4801 agcgtgcctt tgttcactgg tacgtgggtg aggggatgga ggaaggcgag ttttcagagg
     4861 cccgtgaaga tatggctgcc cttgagaagg attatgagga ggttggtgtg gattctgttg
     4921 aaggagaggg tgaggaagaa ggagaggaat actaaggatc caccggatct agataactga
     4981 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     5041 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     5101 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5161 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta
     5221 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     5281 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     5341 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     5401 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     5461 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     5521 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     5581 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     5641 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     5701 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5761 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     5821 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     5881 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     5941 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     6001 ctccgcccat cccgccccta actc
//
