LOCUS       sequence-153298-        6339 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Alpha tubulin labeling with near-infrared fluorescent protein 
            miRFP703.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6339)
  AUTHORS   Shcherbakova DM, Baloban M, Emelyanov AV, Brenowitz M, Guo P, 
            Verkhusha VV
  TITLE     Bright monomeric near-infrared fluorescent proteins as tags and 
            biosensors for multiscale imaging.
  JOURNAL   Nat Commun. 2016 Aug 19;7:12405. doi: 10.1038/ncomms12405.
  PUBMED    27539380
REFERENCE   2  (bases 1 to 6339)
  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: "Nat 
            Commun."; date: "2016-08-19"; volume: "7:12405. doi"; pages: " 
            10.1038/ncomms12405"
COMMENT     Sequence Label: pmiRFP703-Tubulin
FEATURES             Location/Qualifiers
     source          1..6339
                     /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"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      601..610
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             607..1551
                     /codon_start=1
                     /product="bright monomeric near-infrared fluorescent 
                     protein engineered from a bacterial phytochrome, with an 
                     emission maximum of 703 nm (Shcherbakova et al., 2016)"
                     /label=miRFP703
                     /label=ORF frame 1
                     /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH
                     RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE
                     YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG
                     YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSLVPQMARQLYVRQRVRVLVDVT
                     YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPIHLQFLKDMGVRATLAVSLVVGGKLWGLV
                     VCHHYLPRFIRFELRAICKRLAERIATRITALES"
     polyA_signal    3127..3248
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3164..3183)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3218..3237
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3255..3710)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3392..3413)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3604..3623
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3737..3841
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3843..4200
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3843..4034
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3866..3886)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4051..4186
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4235..5029
                     /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             4235..5029
                     /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(4289..4308)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4899..4918
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5217..5236)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5261..5308
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5637..6225
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6126..6145
                     /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 gctagcggtc
      601 gccaccatgg tagcaggtca tgcctctggc agccccgcat tcgggaccgc ctctcattcg
      661 aattgcgaac atgaagagat ccacctcgcc ggctcgatcc agccgcatgg cgcgcttctg
      721 gtcgtcagcg aacatgatca tcgcgtcatc caggccagcg ccaacgccgc ggaatttctg
      781 aatctcggaa gcgtactcgg cgttccgctc gccgagatcg acggcgatct gttgatcaag
      841 atcctgccgc atctcgatcc caccgccgaa ggcatgccgg tcgcggtgcg ctgccggatc
      901 ggcaatccct ctacggagta ctgcggtctg atgcatcggc ctccggaagg cgggctgatc
      961 atcgaactcg aacgtgccgg cccgtcgatc gatctgtcag gcacgctggc gccggcgctg
     1021 gagcggatcc gcacggcggg ttcactgcgc gcgctgtgcg atgacaccgt gctgctgttt
     1081 cagcagtgca ccggctacga ccgggtgatg gtgtatcgtt tcgatgagca aggccacggc
     1141 ctggtattct ccgagtgcca tgtgcctggg ctcgaatcct atttcggcaa ccgctatccg
     1201 tcgtcgctgg tcccgcagat ggcgcggcag ctgtacgtgc ggcagcgcgt ccgcgtgctg
     1261 gtcgacgtca cctatcagcc ggtgccgctg gagccgcggc tgtcgccgct gaccgggcgc
     1321 gatctcgaca tgtcgggctg cttcctgcgc tcgatgtcgc cgatccatct gcagttcctg
     1381 aaggacatgg gcgtgcgcgc caccctggcg gtgtcgctgg tggtcggcgg caagctgtgg
     1441 ggcctggttg tctgtcacca ttatctgccg cgcttcatcc gtttcgagct gcgggcgatc
     1501 tgcaaacggc tcgccgaaag gatcgcgacg cggatcaccg cgcttgagag cggaggtggt
     1561 ggatcaggtg gaggaggttc tggaggtggt ggatcaggag gaggtggttc tggaggtggt
     1621 ggatcaggag gaagatctcg agtgcgtgag tgcatctcca tccacgttgg ccaggctggt
     1681 gtccagattg gcaatgcctg ctgggagctc tactgcctgg aacacggcat ccagcccgat
     1741 ggccagatgc caagtgacaa gaccattggg ggaggagatg actccttcaa caccttcttc
     1801 agtgagacgg gcgctggcaa gcacgtgccc cgggctgtgt ttgtagactt ggaacccaca
     1861 gtcattgatg aagttcgcac tggcacctac cgccagctct tccaccctga gcagctcatc
     1921 acaggcaagg aagatgctgc caataactat gcccgagggc actacaccat tggcaaggag
     1981 atcattgacc ttgtgttgga ccgaattcgc aagctggctg accagtgcac cggtcttcag
     2041 ggcttcttgg ttttccacag ctttggtggg ggaactggtt ctgggttcac ctccctgctc
     2101 atggaacgtc tctcagttga ttatggcaag aagtccaagc tggagttctc catttaccca
     2161 gcaccccagg tttccacagc tgtagttgag ccctacaact ccatcctcac cacccacacc
     2221 accctggagc actctgattg tgccttcatg gtagacaatg aggccatcta tgacatctgt
     2281 cgtagaaacc tcgatatcga gcgcccaacc tacactaacc ttaaccgcct tattagccag
     2341 attgtgtcct ccatcactgc ttccctgaga tttgatggag ccctgaatgt tgacctgaca
     2401 gaattccaga ccaacctggt gccctacccc cgcatccact tccctctggc cacatatgcc
     2461 cctgtcatct ctgctgagaa agcctaccat gaacagcttt ctgtagcaga gatcaccaat
     2521 gcttgctttg agccagccaa ccagatggtg aaatgtgacc ctcgccatgg taaatacatg
     2581 gcttgctgcc tgttgtaccg tggtgacgtg gttcccaaag atgtcaatgc tgccattgcc
     2641 accatcaaaa ccaagcgcag catccagttt gtggattggt gccccactgg cttcaaggtt
     2701 ggcatcaact accagcctcc cactgtggtg cctggtggag acctggccaa ggtacagaga
     2761 gctgtgtgca tgctgagcaa caccacagcc attgctgagg cctgggctcg cctggaccac
     2821 aagtttgacc tgatgtatgc caagcgtgcc tttgttcact ggtacgtggg tgaggggatg
     2881 gaggaaggcg agttttcaga ggcccgtgaa gatatggctg cccttgagaa ggattatgag
     2941 gaggttggtg tggattctgt tgaaggagag ggtgaggaag aaggagagga atactaagga
     3001 tccaccggat ctagataact gatcataatc agccatacca catttgtaga ggttttactt
     3061 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     3121 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3181 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3241 gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     3301 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     3361 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     3421 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     3481 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     3541 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     3601 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     3661 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     3721 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3781 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     3841 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3901 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3961 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4021 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4081 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     4141 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4201 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4261 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4321 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4381 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     4441 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4501 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4561 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4621 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4681 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4741 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4801 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     4861 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     4921 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4981 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5041 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5101 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5161 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     5221 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     5281 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     5341 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     5401 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     5461 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     5521 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5581 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5641 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5701 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5761 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     5821 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5881 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5941 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6001 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6061 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6121 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6181 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6241 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6301 tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
