LOCUS       sequence-367553-        6330 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes enzyme omega-aminotransferase 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 6330)
  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 6330)
  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-ATA-P(11)4
FEATURES             Location/Qualifiers
     source          1..6330
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(233..250)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(260..277)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             296..318
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    333..357
                     /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(357..376)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(358..376)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(460..479)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        685..762
                     /gene="lacI"
                     /label=lacI promoter
     CDS             763..1845
                     /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(782..801)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2654..2845
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2839..2861)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2947..3086
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3002..3021
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3101..3118)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3272..3860)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3352..3371)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(3971..3990)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3982..4797
                     /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             3982..4797
                     /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(4055..4074)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(4889..5344)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5026..5047)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5238..5257
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(5381..5428)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     5424..5442
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(5495..5512)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 tgtgatcgtc cagacgaaag aaacggccat cccaaacgct cggcacatca taggtcaggt
       61 cgctgtgcat gaaaccttgg tccagcagcg ggatacgcgc ttccgccagc ggaaccagtt
      121 caccctccac ccacgcaatg cctttcgcaa acgggttggt ggtctcggtg ctttccagga
      181 tcgcctgacg cgccgcataa cccgcaaaca ctttgtccat gctcgccata tggctgccgc
      241 gcggcaccag gccgctgctg tgatgatgat gatgatggct gctgcccatg gtatatctcc
      301 ttcttaaagt taaacaaaat tatttctaga ggggaattgt tatccgctca caattcccct
      361 atagtgagtc gtattaattt cgcgggatcg agatctcgat cctctacgcc ggacgcatcg
      421 tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg
      481 atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg
      541 tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca ccattccttg
      601 cggcggcggt gctcaacggc ctcaacctac tacygggctg cttcctaatg caggastcgc
      661 ataagggaga gcgtcgagat cccggacacc atcgaatggc gcaaaacctt tcgcggtatg
      721 gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta
      781 tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag
      841 gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat
      901 tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt
      961 gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc
     1021 gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc
     1081 tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat
     1141 ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta
     1201 tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt
     1261 acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg
     1321 ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact
     1381 cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt
     1441 caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac
     1501 gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg
     1561 gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta
     1621 accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa
     1681 ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga
     1741 aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     1801 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     1861 tgtaagttag ctcactcatt aggcaccggg atctcgaccg atgcccttga gagccttcaa
     1921 cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt
     1981 cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca ttttcggcga
     2041 ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat tcggaatctt
     2101 gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca
     2161 ggccattatc gccggcatgg cggccccacg ggtgcgcatg atcgtgctcc tgtcgttgag
     2221 gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac cgatacgcga
     2281 gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt
     2341 cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct gcaccattat
     2401 gttccggatc tgcatcgcag gatgctgctg gctaccctgt ggaacaccta catctgtatt
     2461 aacgaagcgc tggcattgac cctgagtgat ttttctctgg tcccgccgca tccataccgc
     2521 cagttgttta ccctcacaac gttccagtaa ccgggcatgt tcatcatcag taacccgtat
     2581 cgtgagcatc ctctctcgtt tcatcggtat cattaccccc atgaacagaa atccccctta
     2641 cacggaggca tcagtgacca aacaggaaaa aaccgccctt aacatggccc gctttatcag
     2701 aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg aacaggcaga
     2761 catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc tcgcgcgttt
     2821 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     2881 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     2941 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat
     3001 gcggcatcag agcagattgt actgagagtg caccattgcg gtgtgaaata ccgcacagat
     3061 gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc
     3121 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3181 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3241 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3301 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3361 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3421 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3481 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3541 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3601 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3661 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     3721 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3781 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3841 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3901 ggaacgaaaa ctcacgttaa gggattttgg tcatgaacaa taaaactgtc tgcttacata
     3961 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcttg ctctaggccg
     4021 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     4081 gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt
     4141 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
     4201 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     4261 gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt agaagaatat
     4321 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     4381 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
     4441 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
     4501 cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc
     4561 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     4621 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     4681 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     4741 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaagaa
     4801 ttaattcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4861 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
     4921 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     4981 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     5041 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     5101 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     5161 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     5221 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     5281 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     5341 gcgtcccatt cgccaatccg gatatagttc ctcctttcag caaaaaaccc ctcaagaccc
     5401 gtttagaggc cccaaggggt tatgctagtt attgctcagc ggtggcagca gccaactcag
     5461 cttcctttcg ggctttgtta gcagccggat ctcagtggtg gtggtggtgg tgctcgagtt
     5521 attgctgctc aaattcccac tcgaaacgtt gctggctgcc accgccaccg ctgccaccgc
     5581 caccattacg ctcgttatag tcaatttcaa agctatacgc cgcatcgtag tgcatcgccc
     5641 agtagccgtc ccagattttc ttggtaatcg gaccgatctg gccaccgtta accggcatgc
     5701 catccagggt ggtaatcggc atgatgccac ccgcggtggt gcacatgaaa atctcgtcgc
     5761 aacggtacgc cagttcaacc ggcacaaact caacacgcac ttcaatgccg aacgcttccg
     5821 ccgcgttgat aacgctttta cgggtaacac cttgcagcac gccacgatcc ggggtataca
     5881 gcacaccgtc cttaaccagc acgatgttaa agccgctacc ctcggtcagg tgcgcatcac
     5941 cgtcggtcag aaacgggtag gtcgcgccac gatccgccgc ttcaaacatg ccacgaacca
     6001 ggtcgcccca ttgcaggttc ttcacggtcg gatcaatcgc acccggcgga acacgacgca
     6061 cggtacgcgc caccaccgcg ctgccaccca cacgctgcat atccggctcc ataacccaca
     6121 catacggctg aacaaacatg tacaggttgt tcacgatgtc ttccggacgg gtgccacgaa
     6181 cgcctttcag accacgggta acaatcagct ccacgaacgc atcacggata ccgcttttcg
     6241 caaccatttc caccagaatt tgcttcacct ggtcacgcgg cagcggcaga cgcagacgca
     6301 gcttggtgca gctcgcctcc agacgggtga
//
