LOCUS       sequence-368092-        6189 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization and retention of hyperfolder YFP fluorescent protein in
            the ER; improved targeting efficiency.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6189)
  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 6189)
  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: ER2-hfYFP
FEATURES             Location/Qualifiers
     source          1..6189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(514..1305,1306..1374))
                     /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     1137..1156
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1375..1479)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1554..1573)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1745..2124
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2125..2328
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2279..2299
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2373..2392
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2373..2391
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2550..2571)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(3252..3270)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3253..3270)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3290..3307)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3296..3520
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3566..3994
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3653..3672)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3863..3884
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4003..4023)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4008..4337
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4188..4323
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4250..4269
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4404..5198
                     /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(4458..4477)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5068..5087
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5372..5493
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5409..5428)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5463..5482
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5542..5558)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5542..5558)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5555..5577)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5566..5582
                     /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(5590..5596,5597..5614,5615..5620))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5635..5656
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5773..5790)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(5944..6189,1..343))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6024..6043)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
       61 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
      121 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
      181 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
      241 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
      301 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
      361 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
      421 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
      481 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
      541 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
      601 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
      661 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
      721 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
      781 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
      841 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
      901 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
      961 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     1021 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     1081 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     1141 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     1201 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     1261 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     1321 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     1381 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     1441 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     1501 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc
     1561 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     1621 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
     1681 aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg
     1741 cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
     1801 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     1861 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     1921 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     1981 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     2041 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     2101 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2161 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2221 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2281 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact
     2341 agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa
     2401 gcttggtacc gagctcggat ccactagtaa cggccgccag tgtgctgatg ttgttgtccg
     2461 tccctctcct tctgggtctg ctcggcttgg cggtggctga attcggcgtg agcaagggcg
     2521 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     2581 acaagttcag cgtccgcggc gagggcgagg gcgatgccac caacggcaag ctgaccctga
     2641 agctcatctc caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttag
     2701 gctacggcct gatggtgttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     2761 agtccgccat gcccgaaggc tacgtccagg agcgcaccat ctctttcgag gacgacggtt
     2821 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgtgc
     2881 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     2941 tcaacagcca caacgtctat atcacggccg acaagcagaa gaacggcatc aaggctaact
     3001 tcaagatccg ccacaacgtt gaggacggcg gcgtgcagct cgccgaccac taccagcaga
     3061 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt
     3121 ccgtcctgag caaagacccc aacgagaagc gcgatcacat ggtcctgaag gagagggtga
     3181 ccgccgccgg gattacacat gacatgaacg agctgtacaa gtctagaaag gacgagctgt
     3241 aagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg
     3301 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     3361 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     3421 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     3481 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     3541 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3601 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3661 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3721 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3781 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3841 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3901 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3961 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     4021 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4081 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4141 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4201 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4261 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4321 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     4381 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4441 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4501 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4561 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     4621 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4681 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4741 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4801 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4861 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4921 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4981 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5041 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     5101 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5161 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5221 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5281 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5341 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     5401 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5461 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     5521 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5581 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5641 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5701 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5761 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5821 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5881 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5941 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6001 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6061 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6121 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6181 ctccaagct
//
