LOCUS       sequence-271541-        6342 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human alpha-tubulin labeling with a near-infrared fluorescent 
            protein miRFP720.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6342)
  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 6342)
  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: pmiRFP720-tubulin
FEATURES             Location/Qualifiers
     source          1..6342
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        53..356
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        357..560
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     511..531
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      593..602
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             599..1546
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 720 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP720
                     /note="derived from Rhodopseudomonas palustris RpBphP2"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGSVIAEDRCAEVESKLGLHYPASFIPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPNHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACKRVAERLATQIGVMEE"
     polyA_signal    3122..3243
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3159..3178)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3213..3232
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3250..3705)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3387..3408)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3599..3618
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3732..3836
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3838..4195
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3838..4029
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3861..3881)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4046..4181
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4230..5024
                     /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             4230..5024
                     /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(4284..4303)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4894..4913
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5212..5231)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5256..5303
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5632..6220
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6121..6140
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
       61 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      121 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      181 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      241 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      301 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      361 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      421 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      481 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      541 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgg tcgccaccat
      601 ggcggaagga tccgtcgcca ggcagcctga cctcttgacc tgcgacgatg agccgatcca
      661 tatccccggt gccatccaac cgcatggact gctgctcgcc ctcgccgccg acatgacgat
      721 cgttgccggc agcgacaacc ttcccgaact caccggactg gcgatcggcg ccctgatcgg
      781 ccgctctgcg gccgatgtct tcgactcgga gacgcacaac cgtctgacga tcgccttggc
      841 cgagcccggg gcggccgtcg gagcaccgat cactgtcggc ttcacgatgc gaaaggacgc
      901 aggcttcatc ggctcctggc atcgccatga tcagctcatc ttcctcgagc tcgagcctcc
      961 ccagcgggac gtcgccgagc cgcaggcgtt cttccgccgc accaacagcg ccatccgccg
     1021 cctgcaggcc gccgaaacct tggaaagcgc ctgcgccgcc gcggcgcaag aggtgcggaa
     1081 gattaccggc ttcgatcggg tgatgatcta tcgcttcgcc tccgacttca gcgggtccgt
     1141 gatcgcagag gatcggtgcg ccgaggtcga gtcaaaacta ggcctgcact atcctgcctc
     1201 attcatcccg gcgcaggccc gtcggctcta taccatcaac ccggtacgga tcattcccga
     1261 tatcaattat cggccggtgc cggtcacccc agacctcaat ccggtcaccg ggcggccgat
     1321 tgatcttagc ttcgccatcc tgcgcagcgt ctcgcccaac catctggagt tcatgcgcaa
     1381 cataggcatg cacggcacga tgtcgatctc gattttgcgc ggcgagcgac tgtggggatt
     1441 gatcgtttgc catcaccgaa cgccgtacta cgtcgatctc gatggccgcc aagcctgcaa
     1501 gagggtcgcc gagaggctgg ccactcagat cggcgtgatg gaagagggag gtggtggatc
     1561 aggtggagga ggttctggag gtggtggatc aggaggaggt ggttctggag gtggtggatc
     1621 aggaggaaga tctcgagtgc gtgagtgcat ctccatccac gttggccagg ctggtgtcca
     1681 gattggcaat gcctgctggg agctctactg cctggaacac ggcatccagc ccgatggcca
     1741 gatgccaagt gacaagacca ttgggggagg agatgactcc ttcaacacct tcttcagtga
     1801 gacgggcgct ggcaagcacg tgccccgggc tgtgtttgta gacttggaac ccacagtcat
     1861 tgatgaagtt cgcactggca cctaccgcca gctcttccac cctgagcagc tcatcacagg
     1921 caaggaagat gctgccaata actatgcccg agggcactac accattggca aggagatcat
     1981 tgaccttgtg ttggaccgaa ttcgcaagct ggctgaccag tgcaccggtc ttcagggctt
     2041 cttggttttc cacagctttg gtgggggaac tggttctggg ttcacctccc tgctcatgga
     2101 acgtctctca gttgattatg gcaagaagtc caagctggag ttctccattt acccagcacc
     2161 ccaggtttcc acagctgtag ttgagcccta caactccatc ctcaccaccc acaccaccct
     2221 ggagcactct gattgtgcct tcatggtaga caatgaggcc atctatgaca tctgtcgtag
     2281 aaacctcgat atcgagcgcc caacctacac taaccttaac cgccttatta gccagattgt
     2341 gtcctccatc actgcttccc tgagatttga tggagccctg aatgttgacc tgacagaatt
     2401 ccagaccaac ctggtgccct acccccgcat ccacttccct ctggccacat atgcccctgt
     2461 catctctgct gagaaagcct accatgaaca gctttctgta gcagagatca ccaatgcttg
     2521 ctttgagcca gccaaccaga tggtgaaatg tgaccctcgc catggtaaat acatggcttg
     2581 ctgcctgttg taccgtggtg acgtggttcc caaagatgtc aatgctgcca ttgccaccat
     2641 caaaaccaag cgcagcatcc agtttgtgga ttggtgcccc actggcttca aggttggcat
     2701 caactaccag cctcccactg tggtgcctgg tggagacctg gccaaggtac agagagctgt
     2761 gtgcatgctg agcaacacca cagccattgc tgaggcctgg gctcgcctgg accacaagtt
     2821 tgacctgatg tatgccaagc gtgcctttgt tcactggtac gtgggtgagg ggatggagga
     2881 aggcgagttt tcagaggccc gtgaagatat ggctgccctt gagaaggatt atgaggaggt
     2941 tggtgtggat tctgttgaag gagagggtga ggaagaagga gaggaatact aaggatccac
     3001 cggatctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt
     3061 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     3121 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3181 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3241 ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     3301 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     3361 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     3421 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     3481 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     3541 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     3601 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     3661 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     3721 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3781 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     3841 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3901 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3961 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4021 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4081 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     4141 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     4201 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4261 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4321 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4381 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     4441 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4501 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4561 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4621 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4681 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4741 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4801 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4861 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4921 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4981 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     5041 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5101 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5161 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     5221 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     5281 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     5341 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     5401 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     5461 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     5521 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5581 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5641 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5701 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5761 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5821 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5881 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5941 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6001 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6061 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6121 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6181 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6241 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     6301 tgattctgtg gataaccgta ttaccgccat gcattagtta tt
//
