LOCUS       sequence-353407-        5057 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  YPet circularly permuted at a.a. 229; Binder: SspB that carries two 
            point mutations reducing homodimerization(Y11K/A15E, residue 
            numbering from pdb: 1ou9)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5057)
  AUTHORS   Liu B, Stone OJ, Pablo M, Herron JC, Nogueira AT, Dagliyan O, Grimm 
            JB, Lavis LD, Elston TC, Hahn KM
  TITLE     Biosensors based on peptide exposure show single molecule 
            conformations in live cells.
  JOURNAL   Cell. 2021 Oct 28;184(22):5670-5685.e23. doi: 
            10.1016/j.cell.2021.09.026. Epub 2021 Oct 11.
  PUBMED    34637702
REFERENCE   2  (bases 1 to 5057)
  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"; doi: 
            "10.1016/j.cell.2021.09.026"; journalName: "Cell"; date: 
            "2021-10-28- 28"; volume: "184"; issue: "22"; pages: "5670-5685.e23"
COMMENT     Sequence Label: yPet_cp229-Binder
FEATURES             Location/Qualifiers
     source          1..5057
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(247..266)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    291..338
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      667..1255
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1156..1175
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1430..1733
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1734..1937
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1888..1908
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(2334..2353)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2745..3074
                     /codon_start=1
                     /product="SspB from Haemophilus influenzae, with the 
                     mutations Y11K and A15E to disrupt the natural dimerization
                     of the protein"
                     /label=SspB nano
                     /note="dimerizes with iLID at high affinity to create iLID 
                     nano (Guntas et al., 2015)"
                     /translation="SSPKRPKLLREYYDWLVDNSFTPYLVVDATYLGVNVPVEYVKDGQ
                     IVLNLSASATGNLQLTNDFIQFNARFKGVSRELYIPMGAALAIYARENGDGVMFEPEEI
                     YDELNI"
     polyA_signal    3214..3335
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3251..3270)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3305..3324
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3342..3797)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3479..3500)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3691..3710
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3824..3928
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3930..4287
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3930..4121
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3953..3973)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4138..4273
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(4322..5057,1..59)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             join(4322..5057,1..59)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4376..4395)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4986..5005
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
       61 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
      121 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
      181 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
      241 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
      301 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
      361 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
      421 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
      481 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
      541 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
      601 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
      661 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
      721 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
      781 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
      841 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
      901 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
      961 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     1021 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     1081 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     1141 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     1201 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     1261 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     1321 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat
     1381 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     1441 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     1501 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     1561 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     1621 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     1681 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     1741 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     1801 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     1861 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     1921 tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcttaat taagccacca
     1981 tgatcaccga gggcatggac gagctgtaca agggtgctag tggtggcatg gtgagcaagg
     2041 gcgaagagct gttcaccggc gtggtgccca tcctggtgga gctggacggc gacgtgaacg
     2101 gccacaagtt cagcgtgagc ggcgagggcg agggcgacgc cacctacggc aagctgaccc
     2161 tgaagctgct gtgcaccacc ggcaagctgc ccgtgccctg gcccaccctg gtgaccaccc
     2221 tgggctacgg cgtgcagtgc ttcgcccggt accccgacca catgaagcag cacgacttct
     2281 tcaagagcgc catgcccgag ggctacgtgc aggagcggac catcttcttc aaggacgacg
     2341 gcaactacaa gacccgggcc gaggtgaagt tcgagggcga caccctggtg aaccggatcg
     2401 agctgaaggg catcgacttc aaggaggacg gcaacatcct gggccacaag ctggagtaca
     2461 actacaacag ccacaacgtg tacatcaccg ccgacaagca gaagaacggc atcaaggcca
     2521 acttcaagat ccggcacaac atcgaggacg gcggcgtgca gctggccgac cactaccagc
     2581 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc
     2641 agagcgccct gttcaaggac cccaacgaga agcgggacca catggtgctg ctggagttcc
     2701 tgaccgccgc cggctccgga ctcagatctg agctcggatc catgagctcc ccgaaacgcc
     2761 ctaagctgct gcgtgaatat tacgattggc tggttgataa cagctttacc ccatatctgg
     2821 tggtggatgc cacatacctg ggcgtgaacg tgcccgtgga gtatgtgaaa gacggtcaga
     2881 tcgtgctgaa tctgtctgca agtgcgaccg gcaacctgca actgacaaat gattttatcc
     2941 agttcaacgc ccgctttaag ggcgtgtctc gtgaactgta tatcccgatg ggtgccgctc
     3001 tggccattta cgctcgcgag aacggcgatg gtgtgatgtt cgaaccagaa gaaatctatg
     3061 acgagctgaa tattggtaga taagaattct aagcggccgc gactctagat cataatcagc
     3121 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     3181 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     3241 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     3301 agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat
     3361 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     3421 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     3481 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     3541 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     3601 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     3661 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     3721 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     3781 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3841 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3901 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     3961 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4021 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4081 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4141 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4201 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     4261 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     4321 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4381 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4441 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4501 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4561 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4621 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4681 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4741 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4801 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     4861 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4921 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4981 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5041 cctcgtgctt tacggta
//
