LOCUS       sequence-271524-        6306 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human alpha-tubulin labeling with a near-infrared fluorescent 
            protein emiRFP703 (enhanced miRFP703).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6306)
  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 6306)
  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: pemiRFP703-tubulin
FEATURES             Location/Qualifiers
     source          1..6306
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        116..419
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        420..623
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     574..594
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      650..659
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3149..3270
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3186..3205)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3240..3259
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3277..3732)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3414..3435)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3626..3645
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3759..3863
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3865..4222
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3865..4056
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3888..3908)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4073..4208
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4257..5051
                     /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             4257..5051
                     /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(4311..4330)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4921..4940
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5239..5258)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5283..5330
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5659..6247
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6148..6167
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt
       61 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
      121 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      181 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      241 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      301 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      361 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      421 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      481 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      541 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      601 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag ccaccatggc
      661 ggaaggatcc gtcgccaggc agcctgacct cttgacctgc gaacatgaag agatccacct
      721 cgccggctcg atccagccgc atggcgcgct tctggtcgtc agcgaacatg atcatcgcgt
      781 catccaggcc agcgccaacg ccgcggaatt tctgaatctc ggaagcgtac tcggcgttcc
      841 gctcgccgag atcgacggcg atctgttgat caagatcctg ccgcatctcg atcccaccgc
      901 cgaaggcatg ccggtcgcgg tgcgctgccg gatcggcaat ccctctacgg agtactgcgg
      961 tctgatgcat cggcctccgg aaggcgggct gatcatcgaa ctcgaacgtg ccggcccgtc
     1021 gatcgatctg tcaggcacgc tggcgccggc gctggagcgg atccgcacgg cgggttcact
     1081 gcgcgcgctg tgcgatgaca ccgtgctgct gtttcagcag tgcaccggct acgaccgggt
     1141 gatggtgtat cgtttcgatg agcaaggcca cggcctggta ttctccgagt gccatgtgcc
     1201 tgggctcgaa tcctatttcg gcaaccgcta tccgtcgtcg ctggtcccgc agatggcgcg
     1261 gcagctgtac gtgcggcagc gcgtccgcgt gctggtcgac gtcacctatc agccggtgcc
     1321 gctggagccg cggctgtcgc cgctgaccgg gcgcgatctc gacatgtcgg gctgcttcct
     1381 gcgctcgatg tcgccgatcc atctgcagtt cctgaaggac atgggcgtgc gcgccaccct
     1441 ggcggtgtcg ctggtggtcg gcggcaagct gtggggcctg gttgtctgtc accattatct
     1501 gccgcgcttc atccgtttcg agctgcgggc gatctgcaaa cggctcgccg aaaggatcgc
     1561 gacgcggatc accgcgcttg agggtggatc aggtggaggt tctggaggtg gtggatcagg
     1621 aggaggtggt tctggaggtg gtggatcagg aggaagatct cgagtgcgtg agtgcatctc
     1681 catccacgtt ggccaggctg gtgtccagat tggcaatgcc tgctgggagc tctactgcct
     1741 ggaacacggc atccagcccg atggccagat gccaagtgac aagaccattg ggggaggaga
     1801 tgactccttc aacaccttct tcagtgagac gggcgctggc aagcacgtgc cccgggctgt
     1861 gtttgtagac ttggaaccca cagtcattga tgaagttcgc actggcacct accgccagct
     1921 cttccaccct gagcagctca tcacaggcaa ggaagatgct gccaataact atgcccgagg
     1981 gcactacacc attggcaagg agatcattga ccttgtgttg gaccgaattc gcaagctggc
     2041 tgaccagtgc accggtcttc agggcttctt ggttttccac agctttggtg ggggaactgg
     2101 ttctgggttc acctccctgc tcatggaacg tctctcagtt gattatggca agaagtccaa
     2161 gctggagttc tccatttacc cagcacccca ggtttccaca gctgtagttg agccctacaa
     2221 ctccatcctc accacccaca ccaccctgga gcactctgat tgtgccttca tggtagacaa
     2281 tgaggccatc tatgacatct gtcgtagaaa cctcgatatc gagcgcccaa cctacactaa
     2341 ccttaaccgc cttattagcc agattgtgtc ctccatcact gcttccctga gatttgatgg
     2401 agccctgaat gttgacctga cagaattcca gaccaacctg gtgccctacc cccgcatcca
     2461 cttccctctg gccacatatg cccctgtcat ctctgctgag aaagcctacc atgaacagct
     2521 ttctgtagca gagatcacca atgcttgctt tgagccagcc aaccagatgg tgaaatgtga
     2581 ccctcgccat ggtaaataca tggcttgctg cctgttgtac cgtggtgacg tggttcccaa
     2641 agatgtcaat gctgccattg ccaccatcaa aaccaagcgc agcatccagt ttgtggattg
     2701 gtgccccact ggcttcaagg ttggcatcaa ctaccagcct cccactgtgg tgcctggtgg
     2761 agacctggcc aaggtacaga gagctgtgtg catgctgagc aacaccacag ccattgctga
     2821 ggcctgggct cgcctggacc acaagtttga cctgatgtat gccaagcgtg cctttgttca
     2881 ctggtacgtg ggtgagggga tggaggaagg cgagttttca gaggcccgtg aagatatggc
     2941 tgcccttgag aaggattatg aggaggttgg tgtggattct gttgaaggag agggtgagga
     3001 agaaggagag gaatactaag gatccaccgg atctagataa ctgatcataa tcagccatac
     3061 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     3121 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     3181 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3241 tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt
     3301 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     3361 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     3421 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     3481 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     3541 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     3601 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     3661 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     3721 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3781 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3841 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     3901 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3961 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4021 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4081 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4141 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     4201 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     4261 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     4321 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4381 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     4441 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4501 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4561 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4621 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4681 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4741 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     4801 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4861 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4921 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4981 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     5041 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     5101 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     5161 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     5221 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     5281 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     5341 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     5401 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     5461 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     5521 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5581 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5641 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5701 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5761 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5821 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5881 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5941 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6001 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6061 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6121 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6181 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6241 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6301 tcacat
//
