LOCUS       sequence-225651-        7393 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  codes for a fusion of fluorescent protein mGarnet and human 
            alpha-actin, visualizes human alpha-actinin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7393)
  AUTHORS   Hense A, Prunsche B, Gao P, Ishitsuka Y, Nienhaus K, Nienhaus GU
  TITLE     Monomeric Garnet, a far-red fluorescent protein for live-cell STED 
            imaging.
  JOURNAL   Sci Rep. 2015 Dec 9;5:18006. doi: 10.1038/srep18006.
  PUBMED    26648024
REFERENCE   2  (bases 1 to 7393)
  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: "Sci Rep."; 
            date: "2015-12-9"; volume: "5:18006. doi"; pages: " 
            10.1038/srep18006"
COMMENT     Sequence Label: alpha-actinin-mGarnet
FEATURES             Location/Qualifiers
     source          1..7393
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3316..3996
                     /codon_start=1
                     /product="monomeric variant of the red fluorescent protein 
                     eqFP611 (Kredel et al., 2009)"
                     /label=mRuby
                     /translation="MNSLIKENMRMKVVLEGSVNGHQFKCTGEGEGNPYMGTQTMRIKV
                     IEGGPLPFAFDILATSFMYGSKTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGGVITV
                     MQDTSLEDGCLVYHAQVRGVNFPSNGAVMQKKTKGWEPNTEMMYPADGGLRGYNHMALK
                     VDGGGHLSCSLVTTYRSKKTVGNIKMPGIHAVDRRLERLEESDNEMFVVQREHAVAKFA
                     GLGGG"
     polyA_signal    4121..4242
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4158..4177)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4212..4231
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4249..4704)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4386..4407)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4598..4617
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4731..4835
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4837..5194
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4837..5028
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4860..4880)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5045..5180
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5229..6023
                     /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             5229..6023
                     /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(5283..5302)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5893..5912
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6211..6230)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6255..6302
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6631..7219
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7120..7139
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcgc accatcatgg accattatga ttctcagcaa
      601 accaacgatt acatgcagcc agaagaggac tgggaccggg acctgctcct ggacccggcc
      661 tgggagaagc agcagagaaa gacattcacg gcatggtgta actcccacct ccggaaggcg
      721 gggacacaga tcgagaacat cgaagaggac ttccgggatg gcctgaagct catgctgctg
      781 ctggaggtca tctcaggtga acgcttggcc aagccagagc gaggcaagat gagagtgcac
      841 aagatctcca acgtcaacaa ggccctggat ttcatagcca gcaaaggcgt caaactggtg
      901 tccatcggag ccgaagaaat cgtggatggg aatgtgaaga tgaccctggg catgatctgg
      961 accatcatcc tgcgctttgc catccaggac atctccgtgg aagagacttc agccaaggaa
     1021 gggctgctcc tgtggtgtca gagaaagaca gccccttaca aaaatgtcaa catccagaac
     1081 ttccacataa gctggaagga tggcctcggc ttctgtgctt tgatccaccg acaccggccc
     1141 gagctgattg actacgggaa gctgcggaag gatgatccac tcacaaatct gaatacggct
     1201 tttgacgtgg cagagaagta cctggacatc cccaagatgc tggatgccga agacatcgtt
     1261 ggaactgccc gaccggatga gaaagccatc atgacttacg tgtctagctt ctaccacgcc
     1321 ttctctggag cccagaaggc ggagacagca gccaatcgca tctgcaaggt gttggccgtc
     1381 aaccaggaga acgagcagct tatggaagac tacgagaagc tggccagtga tctgttggag
     1441 tggatccgcc gcacaatccc gtggctggag aaccgggtgc ccgagaacac catgcatgcc
     1501 atgcaacaga agctggagga cttccgggac taccggcgcc tgcacaagcc gcccaaggtg
     1561 caggagaagt gccagctgga gatcaacttc aacacgctgc agaccaagct gcggctcagc
     1621 aaccggcctg ccttcatgcc ctctgagggc aggatggtct cggacatcaa caatgcctgg
     1681 ggctgcctgg agcaggtgga gaagggctat gaggagtggt tgctgaatga gatccggagg
     1741 ctggagcgac tggaccacct ggcagagaag ttccggcaga aggcctccat ccacgaggcc
     1801 tggactgacg gcaaagaggc catgctgcga cagaaggact atgagaccgc caccctctcg
     1861 gagatcaagg ccttgctcaa gaagcatgag gccttcgaga gtgacctggc tgcccaccag
     1921 gaccgtgtgg agcagattgc cgccatcgca caggagctca atgagctgga ctattatgac
     1981 tcacccagtg tcaacgcccg ttgccaaaag atctgtgacc agtgggacaa tctgggggcc
     2041 ctaactcaga agcgaaggga agctctggag cggaccgaga aactgctgga gaccattgac
     2101 cagctgtact tggagtatgc caagcgggct gcacccttca acaactggat ggagggggcc
     2161 atggaggacc tgcaggacac cttcattgtg cacaccattg aggagatcca gggactgacc
     2221 acagcccatg agcagttcaa ggccaccctc cctgatgccg acaaggagcg cctggccatc
     2281 ctgggcatcc acaatgaggt gtccaagatt gtccagacct accacgtcaa tatggcgggc
     2341 accaacccct acacaaccat cacgcctcag gagatcaatg gcaaatggga ccacgtgcgg
     2401 cagctggtgc ctcggaggga ccaagctctg acggaggagc atgcccgaca gcagcacaat
     2461 gagaggctac gcaagcagtt tggagcccag gccaatgtca tcgggccctg gatccagacc
     2521 aagatggagg agatcgggag gatctccatt gagatgcatg ggaccctgga ggaccagctc
     2581 agccacctgc ggcagtatga gaagagcatc gtcaactaca agccaaagat tgatcagctg
     2641 gagggcgacc accagctcat ccaggaggcg ctcatcttcg acaacaagca caccaactac
     2701 accatggagc acatccgtgt gggctgggag cagctgctca ccaccatcgc caggaccatc
     2761 aatgaggtag agaaccagat cctgacccgg gatgccaagg gcatcagcca ggagcagatg
     2821 aatgagttcc gggcctcctt caaccacttt gaccgggatc actccggcac actgggtccc
     2881 gaggagttca aagcctgcct catcagcttg ggttatgata ttggcaacga cccccaggga
     2941 gaagcagaat ttgcccgcat catgagcatt gtggacccca accgcctggg ggtagtgaca
     3001 ttccaggcct tcattgactt catgtcccgc gagacagccg acacagatac agcagaccaa
     3061 gtcatggctt ccttcaagat cctggctggg gacaagaact acattaccat ggacgagctg
     3121 cgccgcgagc tgccacccga ccaggctgag tactgcatcg cgcggatggc cccctacacc
     3181 ggccccgact ccgtgccagg tgctctggac tacatgtcct tctccacggc gctgtacggc
     3241 gagagtgacc tcaagcttcg aattctgcag tcgacggtac cgcgggcccg ggatccaccg
     3301 gtcgccaccc gatcgatgaa cagcctgatc aaagaaaaca tgcggatgaa ggtggtgctg
     3361 gaaggcagcg tgaacggcca ccagttcaag tgcaccggcg agggcgaggg caacccctac
     3421 atgggcaccc agaccatgcg gatcaaagtg atcgagggcg gacctctgcc cttcgccttc
     3481 gacatcctgg ccacatcctt catgtacggc agcaagacct tcatcaagta ccccaagggc
     3541 atccccgatt tcttcaagca gagcttcccc gagggcttca cctgggagag agtgaccaga
     3601 tacgaggacg gcggcgtgat caccgtgatg caggacacca gcctggaaga tggctgcctg
     3661 gtgtaccatg cccaggtcag gggcgtgaat tttcccagca acggcgccgt gatgcagaag
     3721 aaaaccaagg gctgggagcc caacaccgag atgatgtacc ccgctgacgg cggactgaga
     3781 ggctacaacc acatggccct gaaggtggac ggcggagggc acctgagctg cagcctggtg
     3841 accacctacc ggtccaagaa aaccgtgggc aacatcaaga tgcccggcat ccacgccgtg
     3901 gaccggcggc tggaaaggct ggaagagtcc gacaacgaga tgttcgtggt gcagcgggag
     3961 cacgccgtgg ccaagttcgc cggcctgggc ggagggtaag cggccgcgac tctagatcat
     4021 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     4081 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     4141 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     4201 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
     4261 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     4321 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     4381 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     4441 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     4501 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     4561 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     4621 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     4681 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4741 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4801 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     4861 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4921 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4981 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     5041 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     5101 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     5161 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     5221 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     5281 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     5341 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     5401 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     5461 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     5521 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     5581 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     5641 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     5701 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     5761 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     5821 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     5881 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     5941 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     6001 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     6061 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     6121 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     6181 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     6241 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     6301 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     6361 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     6421 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     6481 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     6541 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     6601 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6661 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6721 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     6781 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6841 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6901 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6961 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     7021 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     7081 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     7141 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     7201 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     7261 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     7321 taccgccatg cattagttat taatagtaat caattacggg gtcattagtt catagcccat
     7381 atatggagtt ccg
//
