LOCUS       sequence-343872-        7087 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  de novo thick circular tandem repeat protein trimer formed from 
            8-repeat monomers.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7087)
  AUTHORS   Hallinan JP, Doyle LA, Shen BW, Gewe MM, Takushi B, Kennedy MA, 
            Friend D, Roberts JM, Bradley P, Stoddard BL
  TITLE     Design of functionalised circular tandem repeat proteins with longer
            repeat topologies and enhanced subunit contact surfaces.
  JOURNAL   Commun Biol. 2021 Oct 29;4(1):1240. doi: 10.1038/s42003-021-02766-y.
  PUBMED    34716407
REFERENCE   2  (bases 1 to 7087)
  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: "Commun 
            Biol."; date: "2021-10-29"; volume: "4(1):1240. doi"; pages: " 
            10.1038/s42003-021-02766-y"
COMMENT     Sequence Label: tcTRP24sub8
FEATURES             Location/Qualifiers
     source          1..7087
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(45..63)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        131..235
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(236..304,305..1096)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(454..473)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1267..1855
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1756..1775
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2009..2026
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2041..2180
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2106..2125)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2266..2288
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2282..2473)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4828..4847
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4867..4944)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5150..5169
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5253..5272
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5253..5271
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5272..5296
                     /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             5311..5333
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5352..5369
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5379..5396
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(6841..6859)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6855..6902
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(7077..7082,7083..7087,1..12,13..18))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 tcgatgataa gctgtcaaac atgagaattc ttgaagacga aagggcctcg tgatacgcct
       61 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
      121 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
      181 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
      241 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
      301 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
      361 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
      421 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
      481 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
      541 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
      601 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
      661 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
      721 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
      781 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
      841 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
      901 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
      961 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     1021 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     1081 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     1141 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     1201 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     1261 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     1321 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     1381 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     1441 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     1501 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     1561 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     1621 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     1681 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     1741 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     1801 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     1861 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     1921 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     1981 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     2041 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact
     2101 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     2161 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     2221 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     2281 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     2341 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     2401 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     2461 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac
     2521 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac
     2581 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca
     2641 ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc
     2701 agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca
     2761 gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt
     2821 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa
     2881 ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg
     2941 gccaggaccc aacgctgccc gagatgcgcc gcgtgcggct gctggagatg gcggacgcga
     3001 tggatatgtt ctgccaaggg ttggtttgcg cattcacagt tctccgcaag aattgattgg
     3061 ctccaattct tggagtggtg aatccgttag cgaggtgccg ccggcttcca ttcaggtcga
     3121 ggtggcccgg ctccatgcac cgcgacgcaa cgcggggagg cagacaaggt atagggcggc
     3181 gcctacaatc catgccaacc cgttccatgt gctcgccgag gcggcataaa tcgccgtgac
     3241 gatcagcggt ccaatgatcg aagttaggct ggtaagagcc gcgagcgatc cttgaagctg
     3301 tccctgatgg tcgtcatcta cctgcctgga cagcatggcc tgcaacgcgg gcatcccgat
     3361 gccgccggaa gcgagaagaa tcataatggg gaaggccatc cagcctcgcg tcgcgaacgc
     3421 cagcaagacg tagcccagcg cgtcggccgc catgccggcg ataatggcct gcttctcgcc
     3481 gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa
     3541 taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat
     3601 gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag
     3661 tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct
     3721 caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt
     3781 gcgctcactg cccgctttcc agtcgggaaa ctgtcgtgcc agctgcatta atgaatcggc
     3841 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
     3901 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
     3961 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
     4021 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg
     4081 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     4141 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     4201 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     4261 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     4321 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
     4381 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     4441 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     4501 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
     4561 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
     4621 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     4681 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     4741 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     4801 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     4861 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     4921 ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca
     4981 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
     5041 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     5101 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     5161 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     5221 cgtagaggat cgagatctcg atcccgcgaa attaatacga ctcactatag gggaattgtg
     5281 agcggataac aattcccctc tagaaataat tttgtttaac tttaagaagg agatatacta
     5341 tggctagcag ccatcatcat catcatcata gcagcggcct ggtgccgcgc ggcagctcca
     5401 tgggtaacag cgagctggcg gcgcgtatcc tgatcattct gttccagcaa ctggtggagc
     5461 tggcgcgtct ggcgattgaa agcggcgacg aggaactgct gcgtcgtgtg agcgagtggc
     5521 tggaggaagt tatcaaggat atgcgtcgtg tggttgaaca ggcgctgcgt gagggtaaca
     5581 gcgaactggc ggctcgtatc ctgattattc tgtttcagca actggttgag ctggcgcgtc
     5641 tggcgataga gagcggtgat gaagagttat tacgtcgtgt gagcgaatgg ctggaggaag
     5701 ttattaaaga catgcgtcgt gttgtggagc aggcgctgcg cgaaggtaac agcgaactgg
     5761 cggcacgtat cctgataatt ctgtttcaac aactggtgga gctggcgcgt ctggcgatcg
     5821 agagcggtga tgaagaatta ttacgtcgtg tgagcgagtg gctggaggaa gttataaaag
     5881 acatgcgtcg tgtggtggag caggcgctgc gtgaaggtaa cagcgaactg gcggcgcgta
     5941 tcctgataat tctgttccaa caactggttg agctggcgcg tctggcgata gaaagcggtg
     6001 atgaagagct tttacgtcgt gtgagcgagt ggctggagga agttataaag gacatgcgtc
     6061 gtgttgttga gcaggcgctg cgcgagggta acagcgaact ggcggcccgt atcctgataa
     6121 ttctgttcca gcaactggtg gagctggcgc gtctggcgat agaatctggt gatgaagagt
     6181 tacttcgtcg tgtgagcgag tggctggagg aagttataaa ggatatgcgt cgtgttgtgg
     6241 aacaggcgct gcgcgagggt aacagcgaac tggcggcacg tatcctgatc attctgtttc
     6301 agcaactggt tgagctggcg cgtctggcga tagaaagtgg tgatgaagag cttcttcgtc
     6361 gtgtgagcga gtggctggag gaagttataa aggatatgcg tcgtgtagtg gagcaggcgc
     6421 tgcgagaggg taacagcgaa ctggcggcac gtatcctgat tattctgttc cagcaactgg
     6481 tggagctggc gcgtctggcg atagaaagtg gggatgaaga gttactccgt cgtgtgagcg
     6541 agtggctgga ggaagttata aaggatatgc gtcgtgtcgt ggagcaggcg ctgcgggagg
     6601 gtaacagcga actggcggca cgtatcctga taattctgtt ccagcaactg gttgagctgg
     6661 cgcgtctggc gatagaaagt ggcgatgaag agttactacg tcgtgtgagc gagtggctgg
     6721 aggaagttat aaaggatatg cgtcgtgtag tagaacaggc gctgcgcgag ggttaagcgg
     6781 ccgcactcga ggatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
     6841 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     6901 tgctgaaagg aggaactata tccggatatc ccgcaagagg cccggcagta ccggcataac
     6961 caagcctatg cctacagcat ccagggtgac ggtgccgagg atgacgatga gcgcattgtt
     7021 agatttcata cacggtgcct gactgcgtta gcaatttaac tgtgataaac taccgcatta
     7081 aagctta
//
