LOCUS       sequence-254745-        5809 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vivo visualization of microtubule plus ends with near-infrared.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5809)
  AUTHORS   Van Geel O, Cheung S, Gadella TWJ
  TITLE     Combining optogenetics with sensitive FRET imaging to monitor local 
            microtubule manipulations.
  JOURNAL   Sci Rep. 2020 Apr 7;10(1):6034. doi: 10.1038/s41598-020-62874-3.
  PUBMED    32265472
REFERENCE   2  (bases 1 to 5809)
  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: "2020-04-7"; volume: "10(1):6034. doi"; pages: " 
            10.1038/s41598-020-62874-3"
COMMENT     Sequence Label: EB3-iRFP713
FEATURES             Location/Qualifiers
     source          1..5809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      282..291
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1146..1155
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1152..2099
                     /codon_start=1
                     /product="phytochrome-based near-infrared fluorescent 
                     protein, originally termed iRFP (Filonov et al., 2011)"
                     /label=iRFP713
                     /note="name was changed to iRFP713 (Shcherbakova and 
                     Verkhusha, 2013)"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGEVIAEDRCAEVESKLGLHYPASTVPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPVHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     polyA_signal    2233..2354
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2270..2289)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2324..2343
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2361..2816)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2498..2519)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2710..2729
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2843..2947
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2949..3306
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2949..3140
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2972..2992)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3157..3292
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3341..4135
                     /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             3341..4135
                     /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(3395..3414)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4005..4024
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4323..4342)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4367..4414
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4743..5331
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5232..5251
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5506..5809
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 actcagatct cgagctcaag cttcgaattc tgcagtcgac tgccaccatg gccgtcaatg
      301 tgtactccac atctgtgacc agtgaaaatc tgagtcgcca tgatatgctt gcatgggtca
      361 acgactccct gcacctcaac tataccaaga tagaacagct ttgttcaggg gcagcctact
      421 gccagttcat ggacatgctc ttccccggct gtgtgcactt gaggaaagtg aagttccagg
      481 ccaaactaga gcatgaatac atccacaact tcaaggtgct gcaagcagct ttcaagaaga
      541 tgggtgttga caaaatcatt cctgtagaga aattagtgaa aggaaaattc caagataatt
      601 ttgagtttat tcagtggttt aagaaattct ttgacgcaaa ctatgatgga aaggattaca
      661 accctctgct ggcgcggcag ggccaggacg tagcgccacc tcctaaccca ggtgatcaga
      721 tcttcaacaa atccaagaaa ctcattggca cagcagttcc acagaggacg tcccccacag
      781 gcccaaaaaa catgcagacc tctggccggc tgagcaatgt ggcccccccc tgcattctcc
      841 ggaagaatcc tccatcagcc cgaaatggcg gccatgagac tgatgcccaa attcttgaac
      901 tcaaccaaca gctggtggac ttgaagctga cagtggatgg gctggagaag gaacgtgact
      961 tctacttcag caaacttcgt gacatcgagc tcatctgcca ggagcatgaa agtgaaaaca
     1021 gccctgttat ctcaggcatc attggcatcc tctatgccac agaggaagga ttcgcacccc
     1081 ctgaggacga tgagattgaa gagcatcaac aagaagacca ggacgagtac ctggatccac
     1141 cggtcgccac catggcggaa ggatccgtcg ccaggcagcc tgacctcttg acctgcgacg
     1201 atgagccgat ccatatcccc ggtgccatcc aaccgcatgg actgctgctc gccctcgccg
     1261 ccgacatgac gatcgttgcc ggcagcgaca accttcccga actcaccgga ctggcgatcg
     1321 gcgccctgat cggccgctct gcggccgatg tcttcgactc ggagacgcac aaccgtctga
     1381 cgatcgcctt ggccgagccc ggggcggccg tcggagcacc gatcactgtc ggcttcacga
     1441 tgcgaaagga cgcaggcttc atcggctcct ggcatcgcca tgatcagctc atcttcctcg
     1501 agctcgagcc tccccagcgg gacgtcgccg agccgcaggc gttcttccgc cgcaccaaca
     1561 gcgccatccg ccgcctgcag gccgccgaaa ccttggaaag cgcctgcgcc gccgcggcgc
     1621 aagaggtgcg gaagattacc ggcttcgatc gggtgatgat ctatcgcttc gcctccgact
     1681 tcagcggcga agtgatcgca gaggatcggt gcgccgaggt cgagtcaaaa ctaggcctgc
     1741 actatcctgc ctcaaccgtg ccggcgcagg cccgtcggct ctataccatc aacccggtac
     1801 ggatcattcc cgatatcaat tatcggccgg tgccggtcac cccagacctc aatccggtca
     1861 ccgggcggcc gattgatctt agcttcgcca tcctgcgcag cgtctcgccc gtccatctgg
     1921 aattcatgcg caacataggc atgcacggca cgatgtcgat ctcgattttg cgcggcgagc
     1981 gactgtgggg attgatcgtt tgccatcacc gaacgccgta ctacgtcgat ctcgatggcc
     2041 gccaagcctg cgagctagtc gcccaggttc tggcctggca gatcggcgtg atggaagagt
     2101 gtacaagtaa agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt
     2161 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     2221 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     2281 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     2341 atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     2401 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     2461 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     2521 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     2581 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     2641 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     2701 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     2761 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     2821 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     2881 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     2941 gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     3001 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3061 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3121 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3181 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3241 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3301 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3361 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3421 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3481 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
     3541 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3601 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3661 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3721 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     3781 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     3841 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
     3901 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     3961 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4021 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4081 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4141 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4201 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4261 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
     4321 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     4381 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     4441 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     4501 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     4561 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
     4621 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4681 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4741 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4801 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4861 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4921 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4981 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5041 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5101 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5161 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5221 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5281 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5341 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5401 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta
     5461 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     5521 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     5581 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     5641 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     5701 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     5761 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatg
//
