LOCUS       sequence-373443-        6129 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hyperfolder YFP fluorescent protein: cytosolic expression in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6129)
  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 6129)
  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: pcDNA3.1-hfYFP
FEATURES             Location/Qualifiers
     source          1..6129
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(6..25)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        197..576
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        577..780
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     731..751
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     825..844
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        825..843
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(954..975)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(1644..1662)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1645..1662)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1682..1699)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1688..1912
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1958..2386
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2045..2064)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2255..2276
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2395..2415)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2400..2729
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2580..2715
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2642..2661
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2796..3590
                     /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(2850..2869)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3460..3479
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3764..3885
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3801..3820)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3855..3874
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3934..3950)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3934..3950)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3947..3969)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3958..3974
                     /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(3982..3988,3989..4006,4007..4012))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4027..4048
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4165..4182)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4336..4924)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4416..4435)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5095..5886,5887..5955))
                     /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     5718..5737
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5956..6060)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
       61 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
      121 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
      181 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
      241 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      301 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      361 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      421 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      481 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      541 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      601 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      661 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      721 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      781 ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat
      841 agggagaccc aagcttggta ccgagctcgg atccactagt aacggccgcc agtgtgctgg
      901 aattcggcat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      961 agctggacgg cgacgtaaac ggccacaagt tcagcgtccg cggcgagggc gagggcgatg
     1021 ccaccaacgg caagctgacc ctgaagctca tctccaccac cggcaagctg cccgtgccct
     1081 ggcccaccct cgtgaccacc ttaggctacg gcctgatggt gttcgcccgc taccccgacc
     1141 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     1201 ccatctcttt cgaggacgac ggttactaca agacccgcgc cgaggtgaag ttcgagggcg
     1261 acaccctggt gaaccgcatc gtgctgaagg gcatcgactt caaggaggac ggcaacatcc
     1321 tggggcacaa gctggagtac aacttcaaca gccacaacgt ctatatcacg gccgacaagc
     1381 agaagaacgg catcaaggct aacttcaaga tccgccacaa cgttgaggac ggcggcgtgc
     1441 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     1501 acaaccacta cctgagctac cagtccgtcc tgagcaaaga ccccaacgag aagcgcgatc
     1561 acatggtcct gaaggagagg gtgaccgccg ccgggattac acatgacatg aacgagctgt
     1621 acaagtaatc tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca
     1681 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     1741 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     1801 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     1861 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     1921 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     1981 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2041 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2101 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2161 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2221 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2281 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     2341 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     2401 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2461 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2521 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     2581 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     2641 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     2701 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     2761 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     2821 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     2881 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     2941 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     3001 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3061 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3121 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3181 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     3241 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     3301 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     3361 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     3421 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     3481 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     3541 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     3601 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     3661 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     3721 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     3781 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     3841 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     3901 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     3961 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4021 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4081 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4141 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4201 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4261 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4321 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4381 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4441 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4501 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4561 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4621 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4681 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4741 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4801 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4861 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4921 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4981 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5041 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5101 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5161 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5221 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5281 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5341 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5401 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5461 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5521 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5581 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5641 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5701 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5761 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5821 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5881 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5941 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6001 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6061 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     6121 tggtgcact
//
