LOCUS       sequence-343873-        6254 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  de novo thick circular tandem repeat protein capped to prevent 
            multimerization.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6254)
  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 6254)
  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_Cap
FEATURES             Location/Qualifiers
     source          1..6254
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(119..137)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      133..180
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(355..360,361..377,378..383))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(410..428)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        496..600
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(601..669,670..1461)
                     /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(819..838)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1632..2220
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2121..2140
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2374..2391
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2406..2545
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2471..2490)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2631..2653
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2647..2838)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5193..5212
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5232..5309)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5515..5534
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5618..5637
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5618..5636
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5637..5661
                     /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             5676..5698
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5717..5734
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5744..5761
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
ORIGIN
        1 aaggatatgc gtcgtgtaga ggaacaggcg aaacgcgagg gtattatata ataagcggcc
       61 gcactcgagg atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc
      121 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
      181 ctgaaaggag gaactatatc cggatatccc gcaagaggcc cggcagtacc ggcataacca
      241 agcctatgcc tacagcatcc agggtgacgg tgccgaggat gacgatgagc gcattgttag
      301 atttcataca cggtgcctga ctgcgttagc aatttaactg tgataaacta ccgcattaaa
      361 gcttatcgat gataagctgt caaacatgag aattcttgaa gacgaaaggg cctcgtgata
      421 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
      481 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
      541 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
      601 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
      661 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
      721 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
      781 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
      841 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
      901 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
      961 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     1021 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     1081 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     1141 cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     1201 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     1261 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     1321 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     1381 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     1441 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     1501 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     1561 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     1621 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     1681 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     1741 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     1801 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     1861 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     1921 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     1981 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     2041 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     2101 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     2161 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     2221 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     2281 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     2341 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     2401 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt
     2461 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
     2521 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
     2581 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     2641 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc
     2701 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt
     2761 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt
     2821 tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat
     2881 gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg
     2941 gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa
     3001 aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag
     3061 ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt
     3121 ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga
     3181 cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc
     3241 agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac
     3301 ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg agatggcgga
     3361 cgcgatggat atgttctgcc aagggttggt ttgcgcattc acagttctcc gcaagaattg
     3421 attggctcca attcttggag tggtgaatcc gttagcgagg tgccgccggc ttccattcag
     3481 gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg
     3541 gcggcgccta caatccatgc caacccgttc catgtgctcg ccgaggcggc ataaatcgcc
     3601 gtgacgatca gcggtccaat gatcgaagtt aggctggtaa gagccgcgag cgatccttga
     3661 agctgtccct gatggtcgtc atctacctgc ctggacagca tggcctgcaa cgcgggcatc
     3721 ccgatgccgc cggaagcgag aagaatcata atggggaagg ccatccagcc tcgcgtcgcg
     3781 aacgccagca agacgtagcc cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc
     3841 tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt
     3901 ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg
     3961 aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc
     4021 ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag
     4081 gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt
     4141 gcgttgcgct cactgcccgc tttccagtcg ggaaactgtc gtgccagctg cattaatgaa
     4201 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc
     4261 accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc
     4321 aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc
     4381 gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca
     4441 acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca
     4501 accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg
     4561 gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga
     4621 tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac
     4681 agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct
     4741 tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc
     4801 ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta
     4861 atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg
     4921 acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat
     4981 ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca
     5041 atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc
     5101 tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc
     5161 accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt
     5221 actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg
     5281 cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc
     5341 ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa
     5401 tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc
     5461 cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
     5521 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
     5581 tccggcgtag aggatcgaga tctcgatccc gcgaaattaa tacgactcac tataggggaa
     5641 ttgtgagcgg ataacaattc ccctctagaa ataattttgt ttaactttaa gaaggagata
     5701 tactatggct agcagccatc atcatcatca tcatagcagc ggcctggtgc cgcgcggcag
     5761 ctccatgggt ggtaacagcg aggaagcggc gcgtaagctg atcattctgt tccagcaact
     5821 ggtggagaaa gcgcgtaagg cgattgaaag cggcgacgag gaagagctgc gtcgtgtgag
     5881 cgaggaactg gaggaagtta tcaaggatat gcgtcgtgtg gttgaacagg cgctgcgtga
     5941 gggtaacagc gaactggcgg ctcgtatcct gattattctg tttcagcaac tggttgagct
     6001 ggcgcgtctg gcgatagaga gcggtgatga agagttatta cgtcgtgtga gcgaatggct
     6061 ggaggaagtt attaaagaca tgcgtcgtgt tgtggagcag gcgctgcgcg aaggtaacag
     6121 cgaactggcg gcacgtatcc tgataattct gtttcaacaa ctggtggagc tggcgcgtct
     6181 ggcgatcgag agcggtgatg aagaattatt acgtcgtgtg agcgagtggc tggaggaagt
     6241 tataaaagac atgc
//
