LOCUS       sequence-373452-        6189 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization and retention of monomeric 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-mhYFP
FEATURES             Location/Qualifiers
     source          1..6189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(80..99)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        271..650
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        651..854
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     805..825
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     899..918
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        899..917
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1076..1097)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(1778..1796)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1779..1796)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1816..1833)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1822..2046
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2092..2520
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2179..2198)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2389..2410
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2529..2549)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2534..2863
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2714..2849
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2776..2795
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2930..3724
                     /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(2984..3003)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3594..3613
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3898..4019
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3935..3954)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3989..4008
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4068..4084)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4068..4084)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4081..4103)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4092..4108
                     /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(4116..4122,4123..4140,4141..4146))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4161..4182
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4299..4316)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4470..5058)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4550..4569)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5229..6020,6021..6089))
                     /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     5852..5871
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(6090..6189,1..5))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
       61 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
      121 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
      181 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
      241 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
      301 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      361 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      421 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      481 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      541 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      601 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      661 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      721 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      781 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      841 atataagcag agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta
      901 atacgactca ctatagggag acccaagctt ggtaccgagc tcggatccac tagtaacggc
      961 cgccagtgtg ctgatgttgt tgtccgtccc tctccttctg ggtctgctcg gcttggcggt
     1021 ggctgaattc ggcgtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     1081 cgagctggac ggcgacgtaa acggccacaa gttcagcgtc cgcggcgagg gcgagggcga
     1141 tgccaccaac ggcaagctga ccctgaagct catctccacc accggcaagc tgcccgtgcc
     1201 ctggcccacc ctcgtgacca ccttaggcta cggcctgatg gtgttcgccc gctaccccga
     1261 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     1321 caccatctct ttcgaggacg acggttacta caagacccgc gccgaggtga agttcgaggg
     1381 cgacaccctg gtgaaccgca tcgtgctgaa gggcatcgac ttcaaggagg acggcaacat
     1441 cctggggcac aagctggagt acaacttcaa cccccacaac gtctatatca cggccgacaa
     1501 gcagaagaac ggcatcaagg ctaacttcaa gatccgccac aacgttgagg acggcggcgt
     1561 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgatgcc
     1621 cgacaaccac tacctgagct accagtccaa gctgagcaaa gaccccaacg agaagcgcga
     1681 tcacatggtc ctgaaggaga gggtgaccgc cgccgggatt acacatgaca tgaacgagct
     1741 gtacaagtct agaaaggacg agctgtaagg gccctattct atagtgtcac ctaaatgcta
     1801 gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     1861 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     1921 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     1981 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     2041 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     2101 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2161 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2221 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     2281 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     2341 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2401 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     2461 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     2521 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     2581 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     2641 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     2701 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     2761 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     2821 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     2881 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     2941 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     3001 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     3061 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     3121 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     3181 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     3241 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     3301 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     3361 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     3421 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     3481 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     3541 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     3601 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     3661 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     3721 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     3781 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     3841 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     3901 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3961 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     4021 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     4081 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     4141 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     4201 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4261 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4321 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4381 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4441 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4501 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4561 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4621 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4681 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4741 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4801 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4861 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4921 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4981 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5041 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5101 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5161 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5221 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5281 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5341 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5401 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5461 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5521 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5581 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5641 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5701 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5761 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5821 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5881 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5941 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6001 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6061 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6121 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6181 ataggggtt
//
