LOCUS       sequence-225653-        5937 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  codes for a fusion of fluorescent protein mGarnet and alpha-tubulin,
            visualizes alpha-tubulin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5937)
  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 5937)
  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: mGarnet-alpha-tubulin
FEATURES             Location/Qualifiers
     source          1..5937
                     /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             553..1233
                     /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    2737..2858
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2774..2793)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2828..2847
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2865..3320)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3002..3023)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3214..3233
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3347..3451
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3476..3496)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3520..3730
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3542..3738
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3589..3724
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3773..4567
                     /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             3773..4567
                     /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(3827..3846)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4437..4456
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4755..4774)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4799..4846
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5175..5763
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5664..5683
                     /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 ccggtcgcca ccatgaacag cctgatcaaa gaaaacatgc ggatgaaggt ggtgctggaa
      601 ggcagcgtga acggccacca gttcaagtgc accggcgagg gcgagggcaa cccctacatg
      661 ggcacccaga ccatgcggat caaagtgatc gagggcggac ctctgccctt cgccttcgac
      721 atcctggcca catccttcat gtacggcagc aagaccttca tcaagtaccc caagggcatc
      781 cccgatttct tcaagcagag cttccccgag ggcttcacct gggagagagt gaccagatac
      841 gaggacggcg gcgtgatcac cgtgatgcag gacaccagcc tggaagatgg ctgcctggtg
      901 taccatgccc aggtcagggg cgtgaatttt cccagcaacg gcgccgtgat gcagaagaaa
      961 accaagggct gggagcccaa caccgagatg atgtaccccg ctgacggcgg actgagaggc
     1021 tacaaccaca tggccctgaa ggtggacggc ggagggcacc tgagctgcag cctggtgacc
     1081 acctaccgat ccaagaaaac cgtgggcaac atcaagatgc ccggcatcca cgccgtggac
     1141 cggcggctgg aaaggctgga agagtccgac aacgagatgt tcgtggtgca gcgggagcac
     1201 gccgtggcca agttcgccgg cctgggcgga gggtccggac tcagatctcg agtgcgtgag
     1261 tgcatctcca tccacgttgg ccaggctggt gtccagattg gcaatgcctg ctgggagctc
     1321 tactgcctgg aacacggcat ccagcccgat ggccagatgc caagtgacaa gaccattggg
     1381 ggaggagatg actccttcaa caccttcttc agtgagacgg gcgctggcaa gcacgtgccc
     1441 cgggctgtgt ttgtagactt ggaacccaca gtcattgatg aagttcgcac tggcacctac
     1501 cgccagctct tccaccctga gcagctcatc acaggcaagg aagatgctgc caataactat
     1561 gcccgagggc actacaccat tggcaaggag atcattgacc ttgtgttgga ccgaattcgc
     1621 aagctggctg accagtgcac cggtcttcag ggcttcttgg ttttccacag ctttggtggg
     1681 ggaactggtt ctgggttcac ctccctgctc atggaacgtc tctcagttga ttatggcaag
     1741 aagtccaagc tggagttctc catttaccca gcaccccagg tttccacagc tgtagttgag
     1801 ccctacaact ccatcctcac cacccacacc accctggagc actctgattg tgccttcatg
     1861 gtagacaatg aggccatcta tgacatctgt cgtagaaacc tcgatatcga gcgcccaacc
     1921 tacactaacc ttaaccgcct tattagccag attgtgtcct ccatcactgc ttccctgaga
     1981 tttgatggag ccctgaatgt tgacctgaca gaattccaga ccaacctggt gccctacccc
     2041 cgcatccact tccctctggc cacatatgcc cctgtcatct ctgctgagaa agcctaccat
     2101 gaacagcttt ctgtagcaga gatcaccaat gcttgctttg agccagccaa ccagatggtg
     2161 aaatgtgacc ctcgccatgg taaatacatg gcttgctgcc tgttgtaccg tggtgacgtg
     2221 gttcccaaag atgtcaatgc tgccattgcc accatcaaaa ccaagcgcag catccagttt
     2281 gtggattggt gccccactgg cttcaaggtt ggcatcaact accagcctcc cactgtggtg
     2341 cctggtggag acctggccaa ggtacagaga gctgtgtgca tgctgagcaa caccacagcc
     2401 attgctgagg cctgggctcg cctggaccac aagtttgacc tgatgtatgc caagcgtgcc
     2461 tttgttcact ggtacgtggg tgaggggatg gaggaaggcg agttttcaga ggcccgtgaa
     2521 gatatggctg cccttgagaa ggattatgag gaggttggtg tggattctgt tgaaggagag
     2581 ggtgaggaag aaggagagga atactaagga tccaccggat ctagataact gatcataatc
     2641 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     2701 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     2761 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     2821 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
     2881 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     2941 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     3001 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     3061 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     3121 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     3181 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     3241 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     3301 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3361 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3421 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     3481 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3541 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3601 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3661 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3721 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     3781 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     3841 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     3901 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     3961 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4021 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4081 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4141 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4201 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4261 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     4321 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4381 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4441 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4501 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4561 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4621 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4681 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     4741 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     4801 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     4861 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     4921 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     4981 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     5041 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5101 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5161 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5221 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5281 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5341 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5401 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5461 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5521 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5581 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5641 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5701 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5761 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5821 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag
     5881 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccg
//
