LOCUS       sequence-368234-        7578 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hyperfolder YFP, triple fusion: cytosolic expression in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7578)
  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 7578)
  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: pcDNA3.1-3xhfYFP
FEATURES             Location/Qualifiers
     source          1..7578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      944..953
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3131..3149)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3132..3149)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3169..3186)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3175..3399
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3445..3873
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3532..3551)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3742..3763
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3882..3902)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3887..4216
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4067..4202
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4129..4148
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4283..5077
                     /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(4337..4356)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4947..4966
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5251..5372
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5288..5307)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5342..5361
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5421..5437)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5421..5437)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5434..5456)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5445..5461
                     /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(5469..5475,5476..5493,5494..5499))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5514..5535
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5652..5669)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5823..6411)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5903..5922)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6582..7373,7374..7442))
                     /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     7205..7224
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7443..7547)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttcgccacca tggtgagcaa
      961 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     1021 cggccacaag ttcagcgtcc gcggcgaggg cgagggcgat gccaccaacg gcaagctgac
     1081 cctgaagctc atctccacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1141 cttaggctac ggcctgatgg tgttcgcccg ctaccccgac cacatgaagc agcacgactt
     1201 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatctctt tcgaggacga
     1261 cggttactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     1321 cgtgctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     1381 caacttcaac agccacaacg tctatatcac ggccgacaag cagaagaacg gcatcaaggc
     1441 taacttcaag atccgccaca acgttgagga cggcggcgtg cagctcgccg accactacca
     1501 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta
     1561 ccagtccgtc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgaaggagag
     1621 ggtgaccgcc gccgggatta cacatgacat gaacgagctg tacaagctta agatggtgag
     1681 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     1741 aaacggccac aagttcagcg tccgcggcga gggcgagggc gatgccacca acggcaagct
     1801 gaccctgaag ctcatctcca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     1861 caccttaggc tacggcctga tggtgttcgc ccgctacccc gaccacatga agcagcacga
     1921 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct ctttcgagga
     1981 cgacggttac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     2041 catcgtgctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     2101 gtacaacttc aacagccaca acgtctatat cacggccgac aagcagaaga acggcatcaa
     2161 ggctaacttc aagatccgcc acaacgttga ggacggcggc gtgcagctcg ccgaccacta
     2221 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     2281 ctaccagtcc gtcctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgaagga
     2341 gagggtgacc gccgccggga ttacacatga catgaacgag ctgtacaagg ctagcatggt
     2401 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     2461 cgtaaacggc cacaagttca gcgtccgcgg cgagggcgag ggcgatgcca ccaacggcaa
     2521 gctgaccctg aagctcatct ccaccaccgg caagctgccc gtgccctggc ccaccctcgt
     2581 gaccacctta ggctacggcc tgatggtgtt cgcccgctac cccgaccaca tgaagcagca
     2641 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tctctttcga
     2701 ggacgacggt tactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     2761 ccgcatcgtg ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     2821 ggagtacaac ttcaacagcc acaacgtcta tatcacggcc gacaagcaga agaacggcat
     2881 caaggctaac ttcaagatcc gccacaacgt tgaggacggc ggcgtgcagc tcgccgacca
     2941 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     3001 gagctaccag tccgtcctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgaa
     3061 ggagagggtg accgccgccg ggattacaca tgacatgaac gagctgtaca agtaatctag
     3121 agggccctat tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc
     3181 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     3241 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     3301 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     3361 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     3421 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3481 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3541 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3601 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3661 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3721 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3781 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     3841 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     3901 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3961 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4021 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4081 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4141 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4201 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     4261 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4321 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4381 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4441 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     4501 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4561 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4621 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4681 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4741 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4801 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4861 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4921 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4981 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5041 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     5101 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5161 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5221 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     5281 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5341 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     5401 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     5461 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     5521 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5581 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5641 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5701 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5761 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5821 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5881 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5941 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6001 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6061 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6121 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6181 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6241 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     6301 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6361 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6421 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6481 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6541 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6601 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6661 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6721 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6781 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6841 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6901 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6961 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     7021 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7081 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7141 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7201 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7261 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7321 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7381 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7441 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7501 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7561 aaagtgccac ctgacgtc
//
