LOCUS       sequence-477743-        5912 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a de novo 6mer fused to FKBP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5912)
  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 5912)
  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: 6mer-FKBP
FEATURES             Location/Qualifiers
     source          1..5912
                     /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(815..832)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             849..871
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    886..910
                     /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(910..929)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(911..929)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1017..1036)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1242..1319
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1320..2402
                     /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(1339..1358)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3211..3402
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3396..3418)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3504..3643
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3559..3578
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3658..3675)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3829..4417)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3909..3928)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4528..4547)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4539..5354
                     /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             4539..5354
                     /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(4612..4631)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5446..5901)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5583..5604)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5795..5814
                     /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 aatttcagca
      181 gttccacatc aaacaccagg gtcgcatgtg gtggaatgat acccggatgg ccggtcgcgc
      241 catacgcata atccggtgag atggtcagtt ttgcacgctg accaacgctc atctgcgcca
      301 caccttcttc ccagccacga ataacttcct gtttgcccag catgaattta aacggtttgt
      361 tgcgatcgcg gctgctatca aactttttac catcttccag catacctgta taatgcacaa
      421 cgcaggtttg gccgcgtttc ggaaaggtac gaccatcgcc cgggctgatg gtttcaacct
      481 gcacacccga tccaccactt cctccgcttc ccccagaacc accgctacca ccagaaaagc
      541 taaggcttaa gctgacgcct tcaatatccg caatttccag gcacacttta aacacttcca
      601 gtttttcgac gctaacagac aggctaatgc tctggccttt ttccgccagc tttttaaccg
      661 cttcttttaa aatttccgcg acttccactt tcttcaggct gaagctcaag ctcagttcgc
      721 tcgcatccac gctaagaatc gcttccatgg cttctttcat atcttctgct ttttcaaagc
      781 tgccgctcac gctcaggctt ttgccgctgc tgccgtgatg gtggtgatga tgaccgcgca
      841 tatgtatatc tccttcttaa agttaaacaa aattatttct agaggggaat tgttatccgc
      901 tcacaattcc cctatagtga gtcgtattaa tttcgcggga tcgagatcga tctcgatcct
      961 ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg ctggcgccta
     1021 tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca tgagcgcttg
     1081 tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg ccatctcctt
     1141 gcatgcacca ttccttgcgg cggcggtgct caacggcctc aacctactac tgggctgctt
     1201 cctaatgcag gagtcgcata agggagagcg tcgagatccc ggacaccatc gaatggcgca
     1261 aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg gtggtgaatg
     1321 tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat cagaccgttt
     1381 cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg
     1441 cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg ggcaaacagt
     1501 cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg caaattgtcg
     1561 cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac
     1621 gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg
     1681 ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa gctgcctgca
     1741 ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac agtattattt
     1801 tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg ggtcaccagc
     1861 aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct
     1921 ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga
     1981 gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg
     2041 cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg
     2101 ggctgcgcgt tggtgcggac atctcggtag tgggatacga cgataccgaa gacagctcat
     2161 gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg
     2221 tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg
     2281 tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg
     2341 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     2401 gagcgcaacg caattaatgt aagttagctc actcattagg caccgggatc tcgaccgatg
     2461 cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc
     2521 gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc
     2581 tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt
     2641 gcggtattcg gaatcttgca cgccctcgct caagccttcg tcactggtcc cgccaccaaa
     2701 cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg ccccacgggt gcgcatgatc
     2761 gtgctcctgt cgttgaggac ccggctaggc tggcggggtt gccttactgg ttagcagaat
     2821 gaatcaccga tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga
     2881 gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac gcggaagtca
     2941 gcgccctgca ccattatgtt ccggatctgc atcgcaggat gctgctggct accctgtgga
     3001 acacctacat ctgtattaac gaagcgctgg cattgaccct gagtgatttt tctctggtcc
     3061 cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg ggcatgttca
     3121 tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca tcggtatcat tacccccatg
     3181 aacagaaatc ccccttacac ggaggcatca gtgaccaaac aggaaaaaac cgcccttaac
     3241 atggcccgct ttatcagaag ccagacatta acgcttctgg agaaactcaa cgagctggac
     3301 gcggatgaac aggcagacat ctgtgaatcg cttcacgacc acgctgatga gctttaccgc
     3361 agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     3421 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     3481 gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg
     3541 tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac cattgcggtg
     3601 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc
     3661 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3721 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3781 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3841 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3901 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3961 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4021 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4081 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4141 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4201 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4261 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4321 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4381 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4441 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgaacaataa
     4501 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa
     4561 cgtcttgctc taggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
     4621 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg
     4681 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     4741 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
     4801 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccgggaaa acagcattcc
     4861 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
     4921 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     4981 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     5041 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat
     5101 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     5161 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     5221 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     5281 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     5341 atgagttttt ctaagaatta attcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5401 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta
     5461 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     5521 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     5581 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     5641 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     5701 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     5761 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     5821 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     5881 atgcgccgct acagggcgcg tcccattcgc ca
//
