LOCUS       sequence-367552-        6249 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes enzyme tyrosinase linked to P(11)4 self-assembling peptide 
            via a GS-linker sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6249)
  AUTHORS   Shanbhag BK, Liu C, Pradeep GC, Younas T, Hu KKY, Fulcher AJ, Struwe
            WB, Steer D, Dumsday G, Harper IS, Kukura P, Haritos VS, He L
  TITLE     Custom Design of Protein Particles as Multifunctional Biomaterials
  JOURNAL   Adv. Funct. Mater. 2022, 32, 2108039
REFERENCE   2  (bases 1 to 6249)
  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: "Adv. Funct.
            Mater. 2022, 32, 2108039"
COMMENT     Sequence Label: pET28a-Tyr-P(11)4
FEATURES             Location/Qualifiers
     source          1..6249
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     150..172
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(166..357)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1166..2248)
                     /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     2210..2229
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2249..2326)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2532..2551
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2635..2654
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2635..2653
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2654..2678
                     /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)."
     RBS             2693..2715
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2734..2751
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2761..2778
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             3748..3765
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3818..3836)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3832..3879
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3916..4371
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4003..4022)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4213..4234
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4463..5278)
                     /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             complement(4463..5278)
                     /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     5186..5205
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5270..5289
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5400..5988
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5889..5908
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6142..6159
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(6174..6249,1..64)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(join(6239..6249,1..9))
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
       61 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
      121 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
      181 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga
      241 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa
      301 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga
      361 tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag
      421 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg
      481 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca
      541 gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca
      601 gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg
      661 gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct
      721 tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc
      781 tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggggccg ccatgccggc
      841 gataatggcc tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc
      901 gagggcgtgc aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg
      961 aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat
     1021 gataaagaag acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga
     1081 gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag
     1141 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     1201 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
     1261 gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc
     1321 cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt
     1381 tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta
     1441 ccgagatatc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg
     1501 ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca
     1561 tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa
     1621 tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac
     1681 ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc
     1741 ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga
     1801 catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt
     1861 catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg
     1921 ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca
     1981 gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac
     2041 tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt
     2101 tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa
     2161 cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg
     2221 cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc
     2281 gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc
     2341 tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc
     2401 accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg
     2461 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga
     2521 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg
     2581 atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg aaattaatac
     2641 gactcactat aggggaattg tgagcggata acaattcccc tctagaaata attttgttta
     2701 actttaagaa ggagatatac catgggcagc agccatcatc atcatcatca cagcagcggc
     2761 ctggtgccgc gcggcagcca tatgagcaac aagtaccgtg tgcgtaaaaa cgttctgcac
     2821 ctgaccgaca ccgaaaagcg tgatttcgtg cgtaccgttc tgatcctgaa ggagaaaggt
     2881 atctacgacc gttatattgc gtggcacggt gcggcgggta aatttcacac cccgccgggc
     2941 agcgatcgta acgcggcgca catgagcagc gcgttcctgc cgtggcaccg tgaatacctg
     3001 ctgcgttttg agcgtgacct gcaaagcatc aacccggaag tgaccctgcc gtattgggag
     3061 tgggaaaccg acgcgcagat gcaagatccg agccagagcc aaatttggag cgcggacttc
     3121 atgggtggca acggtaaccc gatcaaggac ttcattgttg ataccggtcc gtttgcggcg
     3181 ggccgttgga ccaccattga tgaacagggt aacccgagcg gtggcctgaa gcgtaacttt
     3241 ggtgcgacca aagaggcgcc gaccctgccg acccgtgacg atgtgctgaa cgcgctgaaa
     3301 atcacccaat atgacacccc gccgtgggat atgaccagcc agaacagctt ccgtaaccaa
     3361 ctggagggtt ttattaacgg tccgcagctg cacaaccgtg tgcaccgttg ggttggtggc
     3421 cagatgggtg tgggtccgac cgcgccgaac gacccggttt tctttctgca ccacgcgaac
     3481 gtggatcgta tctgggcggt ttggcagatc attcaccgta accagaacta ccaaccgatg
     3541 aagaacggtc cgttcggcca aaactttcgt gacccgatgt atccgtggaa caccaccccg
     3601 gaagatgtga tgaaccaccg taaactgggt tacgtttatg atattgagct gcgtaagagc
     3661 aaacgtagca gcggtggcgg tggcagcggt ggcggtggca gccagcaacg tttcgagtgg
     3721 gaatttgagc agcaataata actcgagcac caccaccacc accactgaga tccggctgct
     3781 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     3841 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     3901 ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3961 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     4021 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     4081 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     4141 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     4201 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     4261 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     4321 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     4381 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4441 gtatccgctc atgaattaat tcttagaaaa actcatcgag catcaaatga aactgcaatt
     4501 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
     4561 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
     4621 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     4681 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt
     4741 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     4801 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag
     4861 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     4921 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg
     4981 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     5041 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     5101 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga
     5161 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     5221 ccatgttgga atttaatcgc ggcctagagc aagacgtttc ccgttgaata tggctcataa
     5281 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgac caaaatccct
     5341 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5401 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5461 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5521 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     5581 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5641 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5701 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5761 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5821 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5881 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5941 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6001 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6061 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     6121 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     6181 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta
     6241 caatctgct
//
