LOCUS       sequence-271542-        6338 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human alpha-tubulin labeling with a near-infrared fluorescent 
            protein miRFP680.
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: pmiRFP680-tubulin
FEATURES             Location/Qualifiers
     source          1..6338
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        23..326
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        327..530
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     481..501
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      559..568
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             565..1512
                     /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
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPCHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACKRVAERLATQIGVMEE"
     polyA_signal    3088..3209
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3125..3144)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3179..3198
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3216..3671)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3353..3374)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3565..3584
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3698..3802
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3804..4161
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3804..3995
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3827..3847)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4012..4147
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4196..4990
                     /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             4196..4990
                     /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(4250..4269)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4860..4879
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5178..5197)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5222..5269
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5598..6186
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6087..6106
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
       61 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      121 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      181 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      241 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      301 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      361 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      421 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      481 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
      541 accgtcagat ccgctagcgc caccatggcg gaaggctccg tcgccaggca gcctgacctc
      601 ttgacctgcg acgatgagcc gatccatatc cccggtgcca tccaaccgca tggactgctg
      661 ctcgccctcg ccgccgacat gacgatcgtt gccggcagcg acaaccttcc cgaactcacc
      721 ggactggcga tcggcgccct gatcggccgc tctgcggccg atgtcttcga ctcggagacg
      781 cacaaccgtc tgacgatcgc cttggccgag cccggggcgg ccgtcggagc accgatcact
      841 gtcggcttca cgatgcgaaa ggacgcaggc ttcatcggct cctggcatcg ccatgatcag
      901 ctcatcttcc tcgagctcga gcctccccag cgggacgtcg ccgagccgca ggcgttcttc
      961 cgccgcacca acagcgccat ccgccgcctg caggccgccg aaaccttgga aagcgcctgc
     1021 gccgccgcgg cgcaagaggt gcggaagatt accggcttcg atcgggtgat gatctatcgc
     1081 ttcgcctccg acttcagcgg cgaagtgatc gcagaggatc ggtgcgccga ggtcgagtca
     1141 aaactaggcc tgcactatcc tgcctcaacc gtgccggcgc aggcccgtcg gctctatacc
     1201 atcaacccgg tacggatcat tcccgatatc aattatcggc cggtgccggt caccccagac
     1261 ctcaatccgg tcaccgggcg gccgattgat cttagcttcg ccatcctgcg cagcgtctcg
     1321 ccctgccatc tggagttcat gcgcaacata ggcatgcacg gcacgatgtc gatctcgatt
     1381 ttgcgcggcg agcgactgtg gggattgatc gtttgccatc accgaacgcc gtactacgtc
     1441 gatctcgatg gccgccaagc ctgcaagagg gtcgccgaga ggctggccac tcagatcggc
     1501 gtgatggaag agggaggtgg tggatcaggt ggaggaggtt ctggaggtgg tggatcagga
     1561 ggaggtggtt ctggaggtgg tggatcagga ggaagatctc gagtgcgtga gtgcatctcc
     1621 atccacgttg gccaggctgg tgtccagatt ggcaatgcct gctgggagct ctactgcctg
     1681 gaacacggca tccagcccga tggccagatg ccaagtgaca agaccattgg gggaggagat
     1741 gactccttca acaccttctt cagtgagacg ggcgctggca agcacgtgcc ccgggctgtg
     1801 tttgtagact tggaacccac agtcattgat gaagttcgca ctggcaccta ccgccagctc
     1861 ttccaccctg agcagctcat cacaggcaag gaagatgctg ccaataacta tgcccgaggg
     1921 cactacacca ttggcaagga gatcattgac cttgtgttgg accgaattcg caagctggct
     1981 gaccagtgca ccggtcttca gggcttcttg gttttccaca gctttggtgg gggaactggt
     2041 tctgggttca cctccctgct catggaacgt ctctcagttg attatggcaa gaagtccaag
     2101 ctggagttct ccatttaccc agcaccccag gtttccacag ctgtagttga gccctacaac
     2161 tccatcctca ccacccacac caccctggag cactctgatt gtgccttcat ggtagacaat
     2221 gaggccatct atgacatctg tcgtagaaac ctcgatatcg agcgcccaac ctacactaac
     2281 cttaaccgcc ttattagcca gattgtgtcc tccatcactg cttccctgag atttgatgga
     2341 gccctgaatg ttgacctgac agaattccag accaacctgg tgccctaccc ccgcatccac
     2401 ttccctctgg ccacatatgc ccctgtcatc tctgctgaga aagcctacca tgaacagctt
     2461 tctgtagcag agatcaccaa tgcttgcttt gagccagcca accagatggt gaaatgtgac
     2521 cctcgccatg gtaaatacat ggcttgctgc ctgttgtacc gtggtgacgt ggttcccaaa
     2581 gatgtcaatg ctgccattgc caccatcaaa accaagcgca gcatccagtt tgtggattgg
     2641 tgccccactg gcttcaaggt tggcatcaac taccagcctc ccactgtggt gcctggtgga
     2701 gacctggcca aggtacagag agctgtgtgc atgctgagca acaccacagc cattgctgag
     2761 gcctgggctc gcctggacca caagtttgac ctgatgtatg ccaagcgtgc ctttgttcac
     2821 tggtacgtgg gtgaggggat ggaggaaggc gagttttcag aggcccgtga agatatggct
     2881 gcccttgaga aggattatga ggaggttggt gtggattctg ttgaaggaga gggtgaggaa
     2941 gaaggagagg aatactaagg atccaccgga tctagataac tgatcataat cagccatacc
     3001 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     3061 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     3121 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     3181 ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt
     3241 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     3301 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     3361 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     3421 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     3481 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     3541 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     3601 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     3661 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3721 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3781 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3841 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3901 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3961 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4021 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4081 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     4141 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     4201 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4261 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4321 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     4381 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     4441 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     4501 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4561 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4621 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4681 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     4741 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4801 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4861 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4921 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4981 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     5041 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     5101 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     5161 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     5221 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     5281 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     5341 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     5401 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     5461 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5521 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5581 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5641 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5701 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5761 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5821 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5881 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5941 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6001 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6061 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6121 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6181 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6241 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
     6301 tagttattaa tagtaatcaa ttacggggtc attagttc
//
