LOCUS       sequence-245337-        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"
     regulatory      57..66
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      921..930
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             927..1874
                     /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    2008..2129
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2045..2064)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2099..2118
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2136..2591)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2273..2294)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2485..2504
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2618..2722
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2724..3081
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2724..2915
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2747..2767)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2932..3067
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3116..3910
                     /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             3116..3910
                     /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(3170..3189)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3780..3799
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4098..4117)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4142..4189
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4518..5106
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5007..5026
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5281..5584
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5585..5788
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5739..5759
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 cgctagcgct accggactca gatctcgagc tcaagcttcg aattctgcag tcgactgcca
       61 ccatggccgt caatgtgtac tccacatctg tgaccagtga aaatctgagt cgccatgata
      121 tgcttgcatg ggtcaacgac tccctgcacc tcaactatac caagatagaa cagctttgtt
      181 caggggcagc ctactgccag ttcatggaca tgctcttccc cggctgtgtg cacttgagga
      241 aagtgaagtt ccaggccaaa ctagagcatg aatacatcca caacttcaag gtgctgcaag
      301 cagctttcaa gaagatgggt gttgacaaaa tcattcctgt agagaaatta gtgaaaggaa
      361 aattccaaga taattttgag tttattcagt ggtttaagaa attctttgac gcaaactatg
      421 atggaaagga ttacaaccct ctgctggcgc ggcagggcca ggacgtagcg ccacctccta
      481 acccaggtga tcagatcttc aacaaatcca agaaactcat tggcacagca gttccacaga
      541 ggacgtcccc cacaggccca aaaaacatgc agacctctgg ccggctgagc aatgtggccc
      601 ccccctgcat tctccggaag aatcctccat cagcccgaaa tggcggccat gagactgatg
      661 cccaaattct tgaactcaac caacagctgg tggacttgaa gctgacagtg gatgggctgg
      721 agaaggaacg tgacttctac ttcagcaaac ttcgtgacat cgagctcatc tgccaggagc
      781 atgaaagtga aaacagccct gttatctcag gcatcattgg catcctctat gccacagagg
      841 aaggattcgc accccctgag gacgatgaga ttgaagagca tcaacaagaa gaccaggacg
      901 agtacctgga tccaccggtc gccaccatgg cggaaggatc cgtcgccagg cagcctgacc
      961 tcttgacctg cgacgatgag ccgatccata tccccggtgc catccaaccg catggactgc
     1021 tgctcgccct cgccgccgac atgacgatcg ttgccggcag cgacaacctt cccgaactca
     1081 ccggactggc gatcggcgcc ctgatcggcc gctctgcggc cgatgtcttc gactcggaga
     1141 cgcacaaccg tctgacgatc gccttggccg agcccggggc ggccgtcgga gcaccgatca
     1201 ctgtcggctt cacgatgcga aaggacgcag gcttcatcgg ctcctggcat cgccatgatc
     1261 agctcatctt cctcgagctc gagcctcccc agcgggacgt cgccgagccg caggcgttct
     1321 tccgccgcac caacagcgcc atccgccgcc tgcaggccgc cgaaaccttg gaaagcgcct
     1381 gcgccgccgc ggcgcaagag gtgcggaaga ttaccggctt cgatcgggtg atgatctatc
     1441 gcttcgcctc cgacttcagc ggcgaagtga tcgcagagga tcggtgcgcc gaggtcgagt
     1501 caaaactagg cctgcactat cctgcctcaa ccgtgccggc gcaggcccgt cggctctata
     1561 ccatcaaccc ggtacggatc attcccgata tcaattatcg gccggtgccg gtcaccccag
     1621 acctcaatcc ggtcaccggg cggccgattg atcttagctt cgccatcctg cgcagcgtct
     1681 cgcccgtcca tctggaattc atgcgcaaca taggcatgca cggcacgatg tcgatctcga
     1741 ttttgcgcgg cgagcgactg tggggattga tcgtttgcca tcaccgaacg ccgtactacg
     1801 tcgatctcga tggccgccaa gcctgcgagc tagtcgccca ggttctggcc tggcagatcg
     1861 gcgtgatgga agagtgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc
     1921 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     1981 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     2041 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     2101 ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt
     2161 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     2221 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     2281 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     2341 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     2401 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     2461 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     2521 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     2581 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2641 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2701 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     2761 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     2821 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2881 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     2941 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3001 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     3061 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     3121 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3181 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3241 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     3301 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     3361 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     3421 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     3481 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     3541 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     3601 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     3661 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     3721 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     3781 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     3841 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     3901 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     3961 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     4021 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     4081 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     4141 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     4201 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     4261 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     4321 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     4381 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4441 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4501 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4561 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4621 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     4681 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4741 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4801 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4861 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4921 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4981 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5041 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5101 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5161 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
     5221 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     5281 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     5341 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     5401 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     5461 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     5521 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     5581 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     5641 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     5701 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     5761 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatc
//
