LOCUS       sequence-477742-        6233 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a de novo 3mer fused to FKBP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6233)
  AUTHORS   Gregorio NE, Li Z, Hoye JW, David Baker D, DeForest CA
  TITLE     Stimuli-triggered formation of de novo-designed protein biomaterials
  JOURNAL   Cell Biomaterials, 2025
REFERENCE   2  (bases 1 to 6233)
  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: "Cell 
            Biomaterials, 2025"
COMMENT     Sequence Label: 3mer-FKBP
FEATURES             Location/Qualifiers
     source          1..6233
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        145..222
                     /gene="lacI"
                     /label=lacI promoter
     CDS             223..1305
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(242..261)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2114..2305
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2299..2321)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2407..2546
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2462..2481
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2561..2578)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2732..3320)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2812..2831)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(3431..3450)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3442..4257
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             3442..4257
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3515..3534)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(4349..4804)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4486..4507)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4698..4717
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(4841..4888)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4884..4902
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4955..4972)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4979..5302)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             complement(5951..5968)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             5985..6007
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    6022..6046
                     /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)."
     primer_bind     complement(6046..6065)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6047..6065)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6153..6172)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca ccattccttg
       61 cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg caggagtcgc
      121 ataagggaga gcgtcgagat cccggacacc atcgaatggc gcaaaacctt tcgcggtatg
      181 gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta
      241 tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag
      301 gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat
      361 tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt
      421 gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc
      481 gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc
      541 tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat
      601 ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta
      661 tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt
      721 acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg
      781 ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact
      841 cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt
      901 caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac
      961 gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg
     1021 gacatctcgg tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta
     1081 accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa
     1141 ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga
     1201 aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     1261 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     1321 tgtaagttag ctcactcatt aggcaccggg atctcgaccg atgcccttga gagccttcaa
     1381 cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt
     1441 cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca ttttcggcga
     1501 ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat tcggaatctt
     1561 gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca
     1621 ggccattatc gccggcatgg cggccccacg ggtgcgcatg atcgtgctcc tgtcgttgag
     1681 gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac cgatacgcga
     1741 gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt
     1801 cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct gcaccattat
     1861 gttccggatc tgcatcgcag gatgctgctg gctaccctgt ggaacaccta catctgtatt
     1921 aacgaagcgc tggcattgac cctgagtgat ttttctctgg tcccgccgca tccataccgc
     1981 cagttgttta ccctcacaac gttccagtaa ccgggcatgt tcatcatcag taacccgtat
     2041 cgtgagcatc ctctctcgtt tcatcggtat cattaccccc atgaacagaa atccccctta
     2101 cacggaggca tcagtgacca aacaggaaaa aaccgccctt aacatggccc gctttatcag
     2161 aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg aacaggcaga
     2221 catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc tcgcgcgttt
     2281 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     2341 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     2401 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat
     2461 gcggcatcag agcagattgt actgagagtg caccattgcg gtgtgaaata ccgcacagat
     2521 gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc
     2581 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2641 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2701 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2761 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2821 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2881 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2941 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3001 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3061 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3121 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     3181 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3241 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3301 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3361 ggaacgaaaa ctcacgttaa gggattttgg tcatgaacaa taaaactgtc tgcttacata
     3421 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcttg ctctaggccg
     3481 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     3541 gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt
     3601 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
     3661 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     3721 gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt agaagaatat
     3781 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     3841 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
     3901 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
     3961 cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc
     4021 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     4081 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     4141 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     4201 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaagaa
     4261 ttaattcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4321 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
     4381 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     4441 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     4501 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     4561 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     4621 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     4681 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     4741 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     4801 gcgtcccatt cgccaatccg gatatagttc ctcctttcag caaaaaaccc ctcaagaccc
     4861 gtttagaggc cccaaggggt tatgctagtt attgctcagc ggtggcagca gccaactcag
     4921 cttcctttcg ggctttgtta gcagccggat ctcagtggtg gtggtggtgg tgctcgagtt
     4981 attccagttt cagcagttcc acatcaaaca ccagagtcgc atgtggtgga atgatgcccg
     5041 gatggccggt cgcgccatac gcataatccg ggctaatggt cagttttgca cgttggccaa
     5101 cgctcatctg ggccacgcct tcttcccagc cgcgaatcac ttcctgtttg cccaacataa
     5161 atttaaacgg tttgttgcga tcgcggctgc tatcaaactt tttaccatct tccagcatac
     5221 cggtgtagtg cacaacacag gtctggccac gtttcggaaa ggtacggcca tcacccgggc
     5281 tgatggtttc tacctgcact ccgcttcctc cggatccacc acttccacca gacccaccgc
     5341 taccaccact acccgattct ttcagtgtgc ggcgcacttc ggttttaaca cgcaacagaa
     5401 tcagcagaat ttcttcaacc tcggtgccgc ttttcatcag cgccagaata atcgccagca
     5461 caataatctg cacgcattct ttgattactt catagcttga accgctttct tttaaggtgc
     5521 gaatgacttc gctgatcaca cgcgccacaa tttctgcgat ttcgtcttca gaggtcccgc
     5581 tccgtttcaa cgcttcgaca atttcagcga caatgatcgc aacgcaaacg caaataatca
     5641 gataatcgga accggattct ttaagcgtgc gaataacttc agaaattacg cgcgcaacaa
     5701 tttccgcgat ctcatcttcg ctcgtgccag aacgtttcag ggcttccacg atttcggcca
     5761 caatacgtgc cacgcagcgg cagatcactt tatattcgct gccgctctct ttcagggtgc
     5821 ggatcacctc gctaatttcg cgcgccactt cttcggcaat ttcatcctcc gaggtaccgc
     5881 tgcgtttcag acgttccact ttttccgcaa cacggcgcgc tttttcttcg cattcatcgc
     5941 ctgagctgcc atgatggtgg tgatgatgac cgcgcatatg tatatctcct tcttaaagtt
     6001 aaacaaaatt atttctagag gggaattgtt atccgctcac aattccccta tagtgagtcg
     6061 tattaatttc gcgggatcga gatcgatctc gatcctctac gccggacgca tcgtggccgg
     6121 catcaccggc gccacaggtg cggttgctgg cgcctatatc gccgacatca ccgatgggga
     6181 agatcgggct cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta tgg
//
