LOCUS       sequence-271521-        6338 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human alpha-tubulin labeling with a near-infrared fluorescent 
            protein miRFP713.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6338)
  AUTHORS   Matlashov ME, Shcherbakova DM, Alvelid J, Baloban M, Pennacchietti 
            F, Shemetov AA, Testa I, Verkhusha VV
  TITLE     A set of monomeric near-infrared fluorescent proteins for multicolor
            imaging across scales.
  JOURNAL   Nat Commun. 2020 Jan 13;11(1):239. doi: 10.1038/s41467-019-13897-6.
  PUBMED    31932632
REFERENCE   2  (bases 1 to 6338)
  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: "2020-01-13"; volume: "11(1):239. doi"; pages: " 
            10.1038/s41467-019-13897-6"
COMMENT     Sequence Label: pmiRFP713-tubulin
FEATURES             Location/Qualifiers
     source          1..6338
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        72..375
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        376..579
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     530..550
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      608..617
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             614..1561
                     /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
                     IVCHHRTPYYVDLDGRQACKRVAERLATQIGVMEE"
     polyA_signal    3137..3258
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3174..3193)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3228..3247
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3265..3720)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3402..3423)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3614..3633
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3747..3851
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3853..4210
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3853..4044
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3876..3896)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4061..4196
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4245..5039
                     /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             4245..5039
                     /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(4299..4318)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4909..4928
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5227..5246)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5271..5318
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5647..6235
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6136..6155
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgccatgca ttagttatta atagtaatca attacggggt cattagttca tagcccatat
       61 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      121 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      181 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      241 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      301 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      361 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      421 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      481 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      541 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
      601 cgctagcgcc accatggcgg aaggctccgt cgccaggcag cctgacctct tgacctgcga
      661 cgatgagccg atccatatcc ccggtgccat ccaaccgcat ggactgctgc tcgccctcgc
      721 cgccgacatg acgatcgttg ccggcagcga caaccttccc gaactcaccg gactggcgat
      781 cggcgccctg atcggccgct ctgcggccga tgtcttcgac tcggagacgc acaaccgtct
      841 gacgatcgcc ttggccgagc ccggggcggc cgtcggagca ccgatcactg tcggcttcac
      901 gatgcgaaag gacgcaggct tcatcggctc ctggcatcgc catgatcagc tcatcttcct
      961 cgagctcgag cctccccagc gggacgtcgc cgagccgcag gcgttcttcc gccgcaccaa
     1021 cagcgccatc cgccgcctgc aggccgccga aaccttggaa agcgcctgcg ccgccgcggc
     1081 gcaagaggtg cggaagatta ccggcttcga tcgggtgatg atctatcgct tcgcctccga
     1141 cttcagcggc gaagtgatcg cagaggatcg gtgcgccgag gtcgagtcaa aactaggcct
     1201 gcactatcct gcctcaaccg tgccggcgca ggcccgtcgg ctctatacca tcaacccggt
     1261 acggatcatt cccgatatca attatcggcc ggtgccggtc accccagacc tcaatccggt
     1321 caccgggcgg ccgattgatc ttagcttcgc catcctgcgc agcgtctcgc ccgtccatct
     1381 ggagttcatg cgcaacatag gcatgcacgg cacgatgtcg atctcgattt tgcgcggcga
     1441 gcgactgtgg ggattgatcg tttgccatca ccgaacgccg tactacgtcg atctcgatgg
     1501 ccgccaagcc tgcaagaggg tcgccgagag gctggccact cagatcggcg tgatggaaga
     1561 gggaggtggt ggatcaggtg gaggaggttc tggaggtggt ggatcaggag gaggtggttc
     1621 tggaggtggt ggatcaggag gaagatctcg agtgcgtgag tgcatctcca tccacgttgg
     1681 ccaggctggt gtccagattg gcaatgcctg ctgggagctc tactgcctgg aacacggcat
     1741 ccagcccgat ggccagatgc caagtgacaa gaccattggg ggaggagatg actccttcaa
     1801 caccttcttc agtgagacgg gcgctggcaa gcacgtgccc cgggctgtgt ttgtagactt
     1861 ggaacccaca gtcattgatg aagttcgcac tggcacctac cgccagctct tccaccctga
     1921 gcagctcatc acaggcaagg aagatgctgc caataactat gcccgagggc actacaccat
     1981 tggcaaggag atcattgacc ttgtgttgga ccgaattcgc aagctggctg accagtgcac
     2041 cggtcttcag ggcttcttgg ttttccacag ctttggtggg ggaactggtt ctgggttcac
     2101 ctccctgctc atggaacgtc tctcagttga ttatggcaag aagtccaagc tggagttctc
     2161 catttaccca gcaccccagg tttccacagc tgtagttgag ccctacaact ccatcctcac
     2221 cacccacacc accctggagc actctgattg tgccttcatg gtagacaatg aggccatcta
     2281 tgacatctgt cgtagaaacc tcgatatcga gcgcccaacc tacactaacc ttaaccgcct
     2341 tattagccag attgtgtcct ccatcactgc ttccctgaga tttgatggag ccctgaatgt
     2401 tgacctgaca gaattccaga ccaacctggt gccctacccc cgcatccact tccctctggc
     2461 cacatatgcc cctgtcatct ctgctgagaa agcctaccat gaacagcttt ctgtagcaga
     2521 gatcaccaat gcttgctttg agccagccaa ccagatggtg aaatgtgacc ctcgccatgg
     2581 taaatacatg gcttgctgcc tgttgtaccg tggtgacgtg gttcccaaag atgtcaatgc
     2641 tgccattgcc accatcaaaa ccaagcgcag catccagttt gtggattggt gccccactgg
     2701 cttcaaggtt ggcatcaact accagcctcc cactgtggtg cctggtggag acctggccaa
     2761 ggtacagaga gctgtgtgca tgctgagcaa caccacagcc attgctgagg cctgggctcg
     2821 cctggaccac aagtttgacc tgatgtatgc caagcgtgcc tttgttcact ggtacgtggg
     2881 tgaggggatg gaggaaggcg agttttcaga ggcccgtgaa gatatggctg cccttgagaa
     2941 ggattatgag gaggttggtg tggattctgt tgaaggagag ggtgaggaag aaggagagga
     3001 atactaagga tccaccggat ctagataact gatcataatc agccatacca catttgtaga
     3061 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     3121 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     3181 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     3241 actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     3301 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     3361 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     3421 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     3481 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     3541 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     3601 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     3661 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     3721 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3781 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3841 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     3901 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3961 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4021 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4081 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     4141 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4201 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4261 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4321 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4381 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     4441 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4501 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4561 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4621 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4681 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4741 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     4801 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4861 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4921 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4981 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5041 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     5101 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     5161 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     5221 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     5281 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     5341 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     5401 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     5461 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     5521 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5581 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5641 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5701 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5761 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5821 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5881 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5941 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6001 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6061 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6121 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6181 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6241 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     6301 tcctgcgtta tcccctgatt ctgtggataa ccgtatta
//
