LOCUS       sequence-368214-        6045 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization of monomeric hyperfolder YFP fluorescent protein to 
            tubulin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6045)
  AUTHORS   Campbell BC, Paez-Segala MG, Looger LL, Petsko GA, Liu CF
  TITLE     Chemically stable fluorescent proteins for advanced microscopy.
  JOURNAL   Nat Methods. 2022 Nov 7. pii: 10.1038/s41592-022-01660-7. doi: 
            10.1038/s41592-022-01660-7.
  PUBMED    36344833
REFERENCE   2  (bases 1 to 6045)
  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 
            Methods. 2022 Nov 7. pii: 10.1038/s41592-022-01660-7. doi: 
            10.1038/s41592-022-01660-7."
COMMENT     Sequence Label: Tubulin-mhYFP
FEATURES             Location/Qualifiers
     source          1..6045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        80..283
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     234..254
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      322..331
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(373..394)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    2548..2669
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2585..2604)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2639..2658
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2676..3131)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2813..2834)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3025..3044
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3158..3262
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3264..3621
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3264..3455
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3287..3307)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3472..3607
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3656..4450
                     /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             3656..4450
                     /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(3710..3729)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4320..4339
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4638..4657)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4682..4729
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5058..5646
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5547..5566
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        join(5821..6045,1..79)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
       61 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      121 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      181 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      241 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      301 gatccgctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
      361 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtccgc
      421 ggcgagggcg agggcgatgc caccaacggc aagctgaccc tgaagctcat ctccaccacc
      481 ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct taggctacgg cctgatggtg
      541 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
      601 ggctacgtcc aggagcgcac catctctttc gaggacgacg gttactacaa gacccgcgcc
      661 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg tgctgaaggg catcgacttc
      721 aaggaggacg gcaacatcct ggggcacaag ctggagtaca acttcaaccc ccacaacgtc
      781 tatatcacgg ccgacaagca gaagaacggc atcaaggcta acttcaagat ccgccacaac
      841 gttgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
      901 ggccccgtgc tgatgcccga caaccactac ctgagctacc agtccaagct gagcaaagac
      961 cccaacgaga agcgcgatca catggtcctg aaggagaggg tgaccgccgc cgggattaca
     1021 catgacatga acgagctgta caagtccgga ctcagatctc gagtgcgtga gtgcatctcc
     1081 atccacgttg gccaggctgg tgtccagatt ggcaatgcct gctgggagct ctactgcctg
     1141 gaacacggca tccagcccga tggccagatg ccaagtgaca agaccattgg gggaggagat
     1201 gactccttca acaccttctt cagtgagacg ggcgctggca agcacgtgcc ccgggctgtg
     1261 tttgtagact tggaacccac agtcattgat gaagttcgca ctggcaccta ccgccagctc
     1321 ttccaccctg agcagctcat cacaggcaag gaagatgctg ccaataacta tgcccgaggg
     1381 cactacacca ttggcaagga gatcattgac cttgtgttgg accgaattcg caagctggct
     1441 gaccagtgca ccggtcttca gggcttcttg gttttccaca gctttggtgg gggaactggt
     1501 tctgggttca cctccctgct catggaacgt ctctcagttg attatggcaa gaagtccaag
     1561 ctggagttct ccatttaccc agcaccccag gtttccacag ctgtagttga gccctacaac
     1621 tccatcctca ccacccacac caccctggag cactctgatt gtgccttcat ggtagacaat
     1681 gaggccatct atgacatctg tcgtagaaac ctcgatatcg agcgcccaac ctacactaac
     1741 cttaaccgcc ttattagcca gattgtgtcc tccatcactg cttccctgag atttgatgga
     1801 gccctgaatg ttgacctgac agaattccag accaacctgg tgccctaccc ccgcatccac
     1861 ttccctctgg ccacatatgc ccctgtcatc tctgctgaga aagcctacca tgaacagctt
     1921 tctgtagcag agatcaccaa tgcttgcttt gagccagcca accagatggt gaaatgtgac
     1981 cctcgccatg gtaaatacat ggcttgctgc ctgttgtacc gtggtgacgt ggttcccaaa
     2041 gatgtcaatg ctgccattgc caccatcaaa accaagcgca gcatccagtt tgtggattgg
     2101 tgccccactg gcttcaaggt tggcatcaac taccagcctc ccactgtggt gcctggtgga
     2161 gacctggcca aggtacagag agctgtgtgc atgctgagca acaccacagc cattgctgag
     2221 gcctgggctc gcctggacca caagtttgac ctgatgtatg ccaagcgtgc ctttgttcac
     2281 tggtacgtgg gtgaggggat ggaggaaggc gagttttcag aggcccgtga agatatggct
     2341 gcccttgaga aggattatga ggaggttggt gtggattctg ttgaaggaga gggtgaggaa
     2401 gaaggagagg aatactaagg atccaccgga tctagataac tgatcataat cagccatacc
     2461 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     2521 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     2581 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     2641 ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt
     2701 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     2761 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     2821 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     2881 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     2941 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     3001 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     3061 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     3121 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3181 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3241 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3301 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3361 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3421 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3481 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3541 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     3601 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     3661 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3721 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3781 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     3841 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     3901 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     3961 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4021 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4081 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4141 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     4201 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4261 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4321 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4381 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4441 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     4501 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     4561 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     4621 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     4681 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     4741 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     4801 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     4861 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     4921 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4981 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5041 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5101 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5161 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5221 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5281 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5341 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5401 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5461 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5521 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5581 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5641 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5701 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
     5761 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     5821 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     5881 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     5941 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     6001 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctg
//
