LOCUS       sequence-368091-        6045 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization of 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-hfYFP
FEATURES             Location/Qualifiers
     source          1..6045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..153
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      221..230
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(272..293)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    2447..2568
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2484..2503)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2538..2557
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2575..3030)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2712..2733)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2924..2943
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3057..3161
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3163..3520
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3163..3354
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3186..3206)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3371..3506
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3555..4349
                     /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             3555..4349
                     /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(3609..3628)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4219..4238
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4537..4556)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4581..4628
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4957..5545
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5446..5465
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5720..6023
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(6024..6045,1..182)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
       61 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      121 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      181 ctggtttagt gaaccgtcag atccgctagc gctaccggtc gccaccatgg tgagcaaggg
      241 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
      301 ccacaagttc agcgtccgcg gcgagggcga gggcgatgcc accaacggca agctgaccct
      361 gaagctcatc tccaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt
      421 aggctacggc ctgatggtgt tcgcccgcta ccccgaccac atgaagcagc acgacttctt
      481 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atctctttcg aggacgacgg
      541 ttactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcgt
      601 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
      661 cttcaacagc cacaacgtct atatcacggc cgacaagcag aagaacggca tcaaggctaa
      721 cttcaagatc cgccacaacg ttgaggacgg cggcgtgcag ctcgccgacc actaccagca
      781 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca
      841 gtccgtcctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctga aggagagggt
      901 gaccgccgcc gggattacac atgacatgaa cgagctgtac aagtccggac tcagatctcg
      961 agtgcgtgag tgcatctcca tccacgttgg ccaggctggt gtccagattg gcaatgcctg
     1021 ctgggagctc tactgcctgg aacacggcat ccagcccgat ggccagatgc caagtgacaa
     1081 gaccattggg ggaggagatg actccttcaa caccttcttc agtgagacgg gcgctggcaa
     1141 gcacgtgccc cgggctgtgt ttgtagactt ggaacccaca gtcattgatg aagttcgcac
     1201 tggcacctac cgccagctct tccaccctga gcagctcatc acaggcaagg aagatgctgc
     1261 caataactat gcccgagggc actacaccat tggcaaggag atcattgacc ttgtgttgga
     1321 ccgaattcgc aagctggctg accagtgcac cggtcttcag ggcttcttgg ttttccacag
     1381 ctttggtggg ggaactggtt ctgggttcac ctccctgctc atggaacgtc tctcagttga
     1441 ttatggcaag aagtccaagc tggagttctc catttaccca gcaccccagg tttccacagc
     1501 tgtagttgag ccctacaact ccatcctcac cacccacacc accctggagc actctgattg
     1561 tgccttcatg gtagacaatg aggccatcta tgacatctgt cgtagaaacc tcgatatcga
     1621 gcgcccaacc tacactaacc ttaaccgcct tattagccag attgtgtcct ccatcactgc
     1681 ttccctgaga tttgatggag ccctgaatgt tgacctgaca gaattccaga ccaacctggt
     1741 gccctacccc cgcatccact tccctctggc cacatatgcc cctgtcatct ctgctgagaa
     1801 agcctaccat gaacagcttt ctgtagcaga gatcaccaat gcttgctttg agccagccaa
     1861 ccagatggtg aaatgtgacc ctcgccatgg taaatacatg gcttgctgcc tgttgtaccg
     1921 tggtgacgtg gttcccaaag atgtcaatgc tgccattgcc accatcaaaa ccaagcgcag
     1981 catccagttt gtggattggt gccccactgg cttcaaggtt ggcatcaact accagcctcc
     2041 cactgtggtg cctggtggag acctggccaa ggtacagaga gctgtgtgca tgctgagcaa
     2101 caccacagcc attgctgagg cctgggctcg cctggaccac aagtttgacc tgatgtatgc
     2161 caagcgtgcc tttgttcact ggtacgtggg tgaggggatg gaggaaggcg agttttcaga
     2221 ggcccgtgaa gatatggctg cccttgagaa ggattatgag gaggttggtg tggattctgt
     2281 tgaaggagag ggtgaggaag aaggagagga atactaagga tccaccggat ctagataact
     2341 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     2401 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     2461 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     2521 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg
     2581 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     2641 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     2701 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     2761 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     2821 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     2881 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     2941 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     3001 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     3061 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3121 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
     3181 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3241 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3301 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3361 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3421 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     3481 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
     3541 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     3601 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     3661 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     3721 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     3781 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     3841 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     3901 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     3961 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4021 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4081 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4141 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4201 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4261 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4321 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     4381 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     4441 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     4501 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
     4561 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
     4621 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     4681 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     4741 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
     4801 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4861 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4921 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4981 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5041 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5101 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5161 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5221 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5281 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5341 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5401 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5461 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5521 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5581 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     5641 ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata
     5701 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     5761 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     5821 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     5881 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     5941 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     6001 tattagtcat cgctattacc atggtgatgc ggttttggca gtaca
//
