LOCUS       sequence-495652-        6671 bp    DNA     circular SYN 17-DEC-2025
DEFINITION  Expression of mScarlet3-S2 fused with Lamin A/C. This construct is 
            used for imaging..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6671)
  AUTHORS   Ding Y, He W, Wang K, Xue F, Zheng K, Zhao S, Qi D, Li T, Xia P, Pan
            L, Chen L, Chen Z, Yuan L, Xu P
  TITLE     A highly photostable monomeric red fluorescent protein for 
            dual-color 3D STED and time-lapse 3D SIM imaging.
  JOURNAL   Nat Methods. 2025 Nov 26. doi: 10.1038/s41592-025-02962-2.
  PUBMED    41299071
REFERENCE   2  (bases 1 to 6671)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2025 Nov 26. doi: 10.1038/s41592-025-02962-2."
COMMENT     Sequence Label: mScarlet3-S2-Lamin A/C
FEATURES             Location/Qualifiers
     source          1..6671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             600..1286
                     /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
                     VSVAQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDVVLKGDIKH
                     ALRLKDGGRYLADFKTTYRAKKPVQMPGAFNIDRKLDITSHNEDYTVVEQYERSVARHS
                     TGGSGGS"
     primer_bind     998..1017
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3459..3580
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3496..3515)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3550..3569
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3587..4042)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3724..3745)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3936..3955
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4069..4173
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4175..4532
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4175..4366
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4198..4218)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4383..4518
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4567..5361
                     /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             4567..5361
                     /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(4621..4640)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5231..5250
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5549..5568)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5593..5640
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5969..6557
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6458..6477
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 tggatagcac cgaggcagtg atcaaggagt tcatgcggtt caaggtgcac atggagggct
      661 ccatgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
      721 cccagaccgc caagctgagg gtgaccaagg gtggccccct gcccttctcc tgggacatcc
      781 tgtcccctca gttcatgtac ggctccaggg ccttcacgaa gcaccccgcc gacatccccg
      841 actactggaa gcagtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
      901 acggcggcgc cgtgtccgtg gcccaggaca cctccctgga ggacggcacc ctgatctaca
      961 aggtgaagct ccgcggcacc aacttccctc ctgacggccc cgtaatgcag aagaagacaa
     1021 tgggctggga agcatccacc gagcggttgt accccgagga cgtcgtgctg aagggcgaca
     1081 ttaagcacgc cctgcgcctg aaggacggcg gccgctacct ggcggacttc aagaccacct
     1141 acagggccaa gaagcccgtg cagatgcccg gcgccttcaa catcgaccgc aagttggaca
     1201 tcacatccca caacgaggac tacaccgtgg tggaacagta cgaacgctcc gtggcccgcc
     1261 actccaccgg cggctccggt ggctcccaga tctcgagctc aagcttcgaa ttcatggaga
     1321 cctcgtccca gcggcgcgcc acccgcagcg gggcgcaggc cagctccact ccgctgtcgc
     1381 ccacccgcat cacccggctg caggagaagg aggacctgca ggagctcaat gatcgcttgg
     1441 cggtctacat cgaccgtgtg cgctcgctgg aaacggagaa cgcagggctg cgccttcgca
     1501 tcaccgagtc tgaagaggtg gtcagccgcg aggtgtccgg catcaaggcc gcctacgagg
     1561 ccgagctcgg ggatgcccgc aagacccttg actcagtagc caaggagcgc gcccgcctgc
     1621 agctggagct gagcaaagtg cgtgaggagt ttaaggagct gaaagcgcgc aataccaaga
     1681 aggagggtga cctgatagct gctcaggctc ggctgaagga cctggaggct ctgctgaact
     1741 ccaaggaggc cgcactgagc actgctctca gtgagaagcg cacgctggag ggcgagctgc
     1801 atgatctgcg gggccaggtg gccaagcttg aggcagccct aggtgaggcc aagaagcaac
     1861 ttcaggatga gatgctgcgg cgggtggatg ctgagaacag gctgcagacc atgaaggagg
     1921 aactggactt ccagaagaac atctacagtg aggagctgcg tgagaccaag cgccgtcatg
     1981 agacccgact ggtggagatt gacaatggga agcagcgtga gtttgagagc cggctggcgg
     2041 atgcgctgca ggaactgcgg gcccagcatg aggaccaggt ggagcagtat aagaaggagc
     2101 tggagaagac ttattctgcc aagctggaca atgccaggca gtctgctgag aggaacagca
     2161 acctggtggg ggctgcccac gaggagctgc agcagtcgcg catccgcatc gacagcctct
     2221 ctgcccagct cagccagctc cagaagcagc tggcagccaa ggaggcaaag cttcgagacc
     2281 tggaggactc actggcccgt gagcgggaca ccagccggcg gctgctggcg gaaaaggagc
     2341 gggagatggc cgagatgcgg gcaaggatgc agcagcagct ggacgagtac caggagcttc
     2401 tggacatcaa gctggccctg gacatggaga tccacgccta ccgcaagctc ttggagggcg
     2461 aggaggagag gctacgcctg tcccccagcc ctacctcgca gcgcagccgt ggccgtgctt
     2521 cctctcactc atcccagaca cagggtgggg gcagcgtcac caaaaagcgc aaactggagt
     2581 ccactgagag ccgcagcagc ttctcacagc acgcacgcac tagcgggcgc gtggccgtgg
     2641 aggaggtgga cgaggagggc aagtttgtcc ggctgcgcaa caagtccaat gaggaccagt
     2701 ccatgggcaa ttggcagatc aagcgccaga atggagatga tcccttgctg acttaccggt
     2761 tcccaccaaa gttcaccctg aaggctgggc aggtggtgac gatctgggct gcaggagctg
     2821 gggccaccca cagcccccct accgacctgg tgtggaaggc acagaacacc tggggctgcg
     2881 ggaacagcct gcgtacggct ctcatcaact ccactgggga agaagtggcc atgcgcaagc
     2941 tggtgcgctc agtgactgtg gttgaggacg acgaggatga ggatggagat gacctgctcc
     3001 atcaccacca cggctcccac tgcagcagct cgggggaccc cgctgagtac aacctgcgct
     3061 cgcgcaccgt gctgtgcggg acctgcgggc agcctgccga caaggcatct gccagcggct
     3121 caggagccca ggtgggcgga cccatctcct ctggctcttc tgcctccagt gtcacggtca
     3181 ctcgcagcta ccgcagtgtg gggggcagtg ggggtggcag cttcggggac aatctggtca
     3241 cccgctccta cctcctgggc aactccagcc cccgaaccca gagcccccag aactgcagca
     3301 tcatgtaagt cgacggtacc gcgggcccgg gatccaccgg atctagataa ctgatcataa
     3361 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     3421 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     3481 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3541 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt
     3601 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     3661 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     3721 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     3781 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     3841 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     3901 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     3961 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     4021 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4081 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4141 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     4201 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4261 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4321 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4381 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4441 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     4501 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     4561 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4621 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4681 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4741 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4801 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4861 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4921 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4981 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     5041 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     5101 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5161 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5221 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5281 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5341 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5401 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5461 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5521 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     5581 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     5641 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     5701 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     5761 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     5821 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5881 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5941 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6001 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6061 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6121 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6181 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6241 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6301 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6361 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6421 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6481 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6541 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6601 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6661 ccgccatgca t
//
