LOCUS       sequence-368148-        6129 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Monomeric 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-mhYFP
FEATURES             Location/Qualifiers
     source          1..6129
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(529..1320,1321..1389))
                     /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     1152..1171
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1390..1494)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1569..1588)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1760..2139
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2140..2343
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2294..2314
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2388..2407
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2388..2406
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2517..2538)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(3207..3225)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3208..3225)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3245..3262)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3251..3475
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3521..3949
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3608..3627)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3818..3839
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3958..3978)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3963..4292
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4143..4278
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4205..4224
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4359..5153
                     /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(4413..4432)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5023..5042
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5327..5448
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5364..5383)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5418..5437
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5497..5513)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5497..5513)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5510..5532)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5521..5537
                     /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(5545..5551,5552..5569,5570..5575))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5590..5611
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5728..5745)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(5899..6129,1..358))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5979..5998)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
       61 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
      121 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
      181 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
      241 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
      301 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
      361 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
      421 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
      481 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
      541 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
      601 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
      661 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
      721 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
      781 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
      841 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
      901 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
      961 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     1021 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     1081 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     1141 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     1201 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     1261 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     1321 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     1381 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     1441 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     1501 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     1561 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     1621 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     1681 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
     1741 cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg
     1801 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     1861 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     1921 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     1981 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     2041 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     2101 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     2161 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     2221 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     2281 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     2341 gctctctggc taactagaga acccactgct tactggctta tcgaaattaa tacgactcac
     2401 tatagggaga cccaagcttg gtaccgagct cggatccact agtaacggcc gccagtgtgc
     2461 tggaattcgg catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     2521 tcgagctgga cggcgacgta aacggccaca agttcagcgt ccgcggcgag ggcgagggcg
     2581 atgccaccaa cggcaagctg accctgaagc tcatctccac caccggcaag ctgcccgtgc
     2641 cctggcccac cctcgtgacc accttaggct acggcctgat ggtgttcgcc cgctaccccg
     2701 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     2761 gcaccatctc tttcgaggac gacggttact acaagacccg cgccgaggtg aagttcgagg
     2821 gcgacaccct ggtgaaccgc atcgtgctga agggcatcga cttcaaggag gacggcaaca
     2881 tcctggggca caagctggag tacaacttca acccccacaa cgtctatatc acggccgaca
     2941 agcagaagaa cggcatcaag gctaacttca agatccgcca caacgttgag gacggcggcg
     3001 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgatgc
     3061 ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac gagaagcgcg
     3121 atcacatggt cctgaaggag agggtgaccg ccgccgggat tacacatgac atgaacgagc
     3181 tgtacaagta atctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga
     3241 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     3301 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     3361 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     3421 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     3481 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     3541 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3601 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3661 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3721 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3781 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3841 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3901 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     3961 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4021 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4081 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4141 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4201 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     4261 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     4321 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4381 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4441 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4501 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4561 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4621 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4681 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4741 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4801 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4861 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     4921 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4981 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5041 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5101 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5161 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5221 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5281 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     5341 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5401 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     5461 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5521 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5581 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5641 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5701 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5761 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5821 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5881 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5941 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6001 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6061 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6121 ggtgtaggt
//
