LOCUS       sequence-477741-        6338 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a de novo 12mer fused to FKBP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6338)
  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 6338)
  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: 12mer-FKBP
FEATURES             Location/Qualifiers
     source          1..6338
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(164..487)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             complement(1241..1258)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1275..1297
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1312..1336
                     /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(1336..1355)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1337..1355)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1443..1462)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1668..1745
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1746..2828
                     /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(1765..1784)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3637..3828
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3822..3844)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3930..4069
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3985..4004
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4084..4101)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4255..4843)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4335..4354)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4954..4973)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4965..5780
                     /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             4965..5780
                     /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(5038..5057)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5872..6327)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6009..6030)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6221..6240
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagttattcc agtttcaaca
      181 gttccacatc aaaaaccaga gtcgcatgtg gcggaatgat acccggatgg ccggtcgcac
      241 catacgcata atccgggcta atggtcagtt ttgcgcgctg acccacgctc atttgcgcca
      301 cgccttcttc ccagccacga atcacttcct gtttgcccag cataaattta aacggtttgt
      361 tgcgatcgcg gctgctatca aactttttac cgtcttccaa cataccggta taatgcacaa
      421 cgcaggtctg gccgcgtttc ggaaacgtac gaccatcacc cgggctgatg gtttccacct
      481 gcacaccgga accaccacta ccacccgaac caccgctacc accagaaccg ccttgttttt
      541 ccagcgcttt caccacttct tcgttgccat gttcacgggc cagatccagc ggggttctgc
      601 catcggaatc gctggtgttt acatctgcgc ccgccaggat cagaaccagt accacacgtt
      661 tatggccgtt ttctgcggcg tggtgcagag gggtgcggcc atctgaatct ttggcgttga
      721 catccgcgcc tttcagcagc agaatcagca ctacttcatc atgtccgttt tccgcggcat
      781 ggtgcaacgg ggtacggcca tcgctgtctt tttcattaac atcggcgcct ttttcgatca
      841 gcagggtcac aacttctgca tgaccatttt cagccgcgtg atgcagcggt gtgcgaccgt
      901 cgctatcctt ttcgttcaca tccgcacccg catcgatcgc cactttaacc acagctgcag
      961 caccagcttc cgccgcatga tgcaaaggag tgcgcccatc actatctttc gcgttcgcat
     1021 ctgcaccctg gttcatggcg ctaatcgcct ggttggccac gctggcttca atcgctttga
     1081 caataacttt aagcactttc acaatggtat ctttatccgg gttcttttcc aggcgttcca
     1141 gcatttcttt cagttccgcc agaatgcgaa tcagttcaat caggctatct tccagctgtt
     1201 tcagcagttc ctttttcttt tcttcatcgc ccgagctacc gtgatggtga tgatgatgac
     1261 cgcgcatatg tatatctcct tcttaaagtt aaacaaaatt atttctagag gggaattgtt
     1321 atccgctcac aattccccta tagtgagtcg tattaatttc gcgggatcga gatcgatctc
     1381 gatcctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg
     1441 cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag
     1501 cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat
     1561 ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc tactactggg
     1621 ctgcttccta atgcaggagt cgcataaggg agagcgtcga gatcccggac accatcgaat
     1681 ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg
     1741 tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga
     1801 ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg
     1861 aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca
     1921 aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa
     1981 ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg
     2041 tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg
     2101 tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg
     2161 cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta
     2221 ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc
     2281 accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg
     2341 ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg
     2401 actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc
     2461 ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg
     2521 agtccgggct gcgcgttggt gcggacatct cggtagtggg atacgacgat accgaagaca
     2581 gctcatgtta tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa
     2641 ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt
     2701 tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc
     2761 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     2821 ggcagtgagc gcaacgcaat taatgtaagt tagctcactc attaggcacc gggatctcga
     2881 ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact
     2941 atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca
     3001 gcgctctggg tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg
     3061 tcgcttgcgg tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc
     3121 accaaacgtt tcggcgagaa gcaggccatt atcgccggca tggcggcccc acgggtgcgc
     3181 atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct tactggttag
     3241 cagaatgaat caccgatacg cgagcgaacg tgaagcgact gctgctgcaa aacgtctgcg
     3301 acctgagcaa caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct ggaaacgcgg
     3361 aagtcagcgc cctgcaccat tatgttccgg atctgcatcg caggatgctg ctggctaccc
     3421 tgtggaacac ctacatctgt attaacgaag cgctggcatt gaccctgagt gatttttctc
     3481 tggtcccgcc gcatccatac cgccagttgt ttaccctcac aacgttccag taaccgggca
     3541 tgttcatcat cagtaacccg tatcgtgagc atcctctctc gtttcatcgg tatcattacc
     3601 cccatgaaca gaaatccccc ttacacggag gcatcagtga ccaaacagga aaaaaccgcc
     3661 cttaacatgg cccgctttat cagaagccag acattaacgc ttctggagaa actcaacgag
     3721 ctggacgcgg atgaacaggc agacatctgt gaatcgcttc acgaccacgc tgatgagctt
     3781 taccgcagct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     3841 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     3901 gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc
     3961 ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccatt
     4021 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc
     4081 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4141 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4201 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4261 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4321 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4381 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4441 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4501 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4561 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4621 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4681 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4741 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4801 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4861 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgaa
     4921 caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac
     4981 gggaaacgtc ttgctctagg ccgcgattaa attccaacat ggatgctgat ttatatgggt
     5041 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga
     5101 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
     5161 cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc
     5221 attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc gggaaaacag
     5281 cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag
     5341 tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg
     5401 tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
     5461 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt
     5521 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
     5581 ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat
     5641 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
     5701 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
     5761 tgctcgatga gtttttctaa gaattaattc atgagcggat acatatttga atgtatttag
     5821 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
     5881 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     5941 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     6001 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     6061 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     6121 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     6181 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     6241 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     6301 cgcttaatgc gccgctacag ggcgcgtccc attcgcca
//
