LOCUS       sequence-368011-        7496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tubulin labeling with blue fluorescent protein mTagBFP2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7496)
  AUTHORS   Papadaki S, Wang X, Wang Y, Zhang H, Jia S, Liu S, Yang M, Zhang D, 
            Jia JM, Koster RW, Namikawa K, Piatkevich KD
  TITLE     Dual-expression system for blue fluorescent protein optimization.
  JOURNAL   Sci Rep. 2022 Jun 17;12(1):10190. doi: 10.1038/s41598-022-13214-0.
  PUBMED    35715437
REFERENCE   2  (bases 1 to 7496)
  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: "Sci Rep."; 
            date: "2022-06-17"; volume: "12(1):10190. doi"; pages: " 
            10.1038/s41598-022-13214-0"
COMMENT     Sequence Label: pTubulin-mTagBFP2
FEATURES             Location/Qualifiers
     source          1..7496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        137..516
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        517..720
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     671..691
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     765..784
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        765..783
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             809..1507
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     promoter        complement(2951..2969)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2952..2969)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2989..3006)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2995..3219
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3265..3693
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3352..3371)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3562..3583
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3702..3722)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3707..4036
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3887..4022
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3949..3968
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4103..4897
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4157..4176)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4767..4786
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5071..5192
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5108..5127)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5162..5181
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5241..5257)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5241..5257)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5254..5276)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5265..5281
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5289..5295,5296..5313,5314..5319))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5334..5355
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5472..5489)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5643..6231)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5723..5742)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6402..7193,7194..7262))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7025..7044
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7263..7367)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7442..7461)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
       61 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
      121 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
      181 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      241 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      301 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      361 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      421 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      481 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      541 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      601 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      661 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      721 ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat
      781 agggagaccc aagcttggta ccgccaccat gagcgagctg attaaggaga acatgcacat
      841 gaagctgtac atggagggca ccgtggacaa ccatcacttc aagtgcacat ccgagggcga
      901 aggcaagccc tacgagggca cccagaccat gagaatcaag gtggtcgagg gcggccctct
      961 ccccttcgcc ttcgacatcc tggctactag cttcctctac ggcagcaaga ccttcatcaa
     1021 ccacacccag ggcatccccg acttcttcaa gcagtccttc cctgagggct tcacatggga
     1081 gagagtcacc acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcctcca
     1141 ggacggctgc ctcatctaca acgtcaagat cagaggggtg aacttcacat ccaacggccc
     1201 tgtgatgcag aagaaaacac tcggctggga ggccttcacc gagacgctgt accccgctga
     1261 cggcggcctg gaaggcagaa acgacatggc cctgaagctc gtgggcggga gccatctgat
     1321 cgcaaacgcc aagaccacat atagatccaa gaaacccgct aagaacctca agatgcctgg
     1381 cgtctactat gtggactaca gactggaaag aatcaaggag gccaacaacg agacctacgt
     1441 cgagcagcac gaggtggcag tggccagata ctgcgacctc cctagcaaac tggggcacaa
     1501 gcttaatgga ggtggtggat caggaggagg tggttctgga ggtggtggat caggaggaag
     1561 atctcgagtg cgtgagtgca tctccatcca cgttggccag gctggtgtcc agattggcaa
     1621 tgcctgctgg gagctctact gcctggaaca cggcatccag cccgatggcc agatgccaag
     1681 tgacaagacc attgggggag gagatgactc cttcaacacc ttcttcagtg agacgggcgc
     1741 tggcaagcac gtgccccggg ctgtgtttgt agacttggaa cccacagtca ttgatgaagt
     1801 tcgcactggc acctaccgcc agctcttcca ccctgagcag ctcatcacag gcaaggaaga
     1861 tgctgccaat aactatgccc gagggcacta caccattggc aaggagatca ttgaccttgt
     1921 gttggaccga attcgcaagc tggctgacca gtgcaccggt cttcagggct tcttggtttt
     1981 ccacagcttt ggtgggggaa ctggttctgg gttcacctcc ctgctcatgg aacgtctctc
     2041 agttgattat ggcaagaagt ccaagctgga gttctccatt tacccagcac cccaggtttc
     2101 cacagctgta gttgagccct acaactccat cctcaccacc cacaccaccc tggagcactc
     2161 tgattgtgcc ttcatggtag acaatgaggc catctatgac atctgtcgta gaaacctcga
     2221 tatcgagcgc ccaacctaca ctaaccttaa ccgccttatt agccagattg tgtcctccat
     2281 cactgcttcc ctgagatttg atggagccct gaatgttgac ctgacagaat tccagaccaa
     2341 cctggtgccc tacccccgca tccacttccc tctggccaca tatgcccctg tcatctctgc
     2401 tgagaaagcc taccatgaac agctttctgt agcagagatc accaatgctt gctttgagcc
     2461 agccaaccag atggtgaaat gtgaccctcg ccatggtaaa tacatggctt gctgcctgtt
     2521 gtaccgtggt gacgtggttc ccaaagatgt caatgctgcc attgccacca tcaaaaccaa
     2581 gcgcagcatc cagtttgtgg attggtgccc cactggcttc aaggttggca tcaactacca
     2641 gcctcccact gtggtgcctg gtggagacct ggccaaggta cagagagctg tgtgcatgct
     2701 gagcaacacc acagccattg ctgaggcctg ggctcgcctg gaccacaagt ttgacctgat
     2761 gtatgccaag cgtgcctttg ttcactggta cgtgggtgag gggatggagg aaggcgagtt
     2821 ttcagaggcc cgtgaagata tggctgccct tgagaaggat tatgaggagg ttggtgtgga
     2881 ttctgttgaa ggagagggtg aggaagaagg agaggaatac taaggatcca ccggatctag
     2941 agggccctat tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc
     3001 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     3061 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     3121 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     3181 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     3241 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3301 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3361 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3421 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3481 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3541 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3601 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     3661 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     3721 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3781 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3841 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3901 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3961 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4021 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     4081 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4141 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4201 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4261 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     4321 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4381 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4441 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4501 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4561 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4621 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4681 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4741 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4801 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4861 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4921 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4981 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5041 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     5101 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5161 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     5221 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     5281 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     5341 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5401 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5461 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5521 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5581 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5641 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5701 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5761 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5821 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5881 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5941 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6001 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6061 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     6121 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6181 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6241 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6301 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6361 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6421 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6481 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6541 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6601 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6661 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6721 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6781 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6841 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6901 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6961 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7021 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7081 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7141 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7201 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7261 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7321 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7381 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca
     7441 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtg
//
