LOCUS       sequence-343860-        6745 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  de novo thick circular tandem repeat protein tetramer formed from 
            6-repeat monomers.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6745)
  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 6745)
  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: tcTRP24sub6
FEATURES             Location/Qualifiers
     source          1..6745
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(43..61)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        129..233
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(234..302,303..1094)
                     /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(452..471)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1265..1853
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1754..1773
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2007..2024
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2039..2178
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2104..2123)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2264..2286
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2280..2471)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4826..4845
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4865..4942)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5148..5167
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5251..5270
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5251..5269
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5270..5294
                     /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             5309..5331
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5350..5367
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5377..5394
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(6497..6515)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6511..6558
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(6733..6738,6739..6745,1..10,11..16))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 gatgataagc tgtcaaacat gagaattctt gaagacgaaa gggcctcgtg atacgcctat
       61 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
      121 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
      181 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
      241 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
      301 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
      361 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
      421 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
      481 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
      541 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
      601 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
      661 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
      721 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag
      781 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
      841 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
      901 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
      961 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1021 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1081 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     1141 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     1201 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     1261 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     1321 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     1381 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     1441 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     1501 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     1561 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     1621 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     1681 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     1741 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     1801 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     1861 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     1921 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     1981 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     2041 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct
     2101 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
     2161 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     2221 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     2281 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
     2341 tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
     2401 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
     2461 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg
     2521 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg
     2581 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact
     2641 cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag
     2701 catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga
     2761 ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg
     2821 cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg
     2881 caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggc
     2941 caggacccaa cgctgcccga gatgcgccgc gtgcggctgc tggagatggc ggacgcgatg
     3001 gatatgttct gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa ttgattggct
     3061 ccaattcttg gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt caggtcgagg
     3121 tggcccggct ccatgcaccg cgacgcaacg cggggaggca gacaaggtat agggcggcgc
     3181 ctacaatcca tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc gccgtgacga
     3241 tcagcggtcc aatgatcgaa gttaggctgg taagagccgc gagcgatcct tgaagctgtc
     3301 cctgatggtc gtcatctacc tgcctggaca gcatggcctg caacgcgggc atcccgatgc
     3361 cgccggaagc gagaagaatc ataatgggga aggccatcca gcctcgcgtc gcgaacgcca
     3421 gcaagacgta gcccagcgcg tcggccgcca tgccggcgat aatggcctgc ttctcgccga
     3481 aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata
     3541 ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga
     3601 cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg
     3661 cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca
     3721 agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc
     3781 gctcactgcc cgctttccag tcgggaaact gtcgtgccag ctgcattaat gaatcggcca
     3841 acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg
     3901 agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt
     3961 ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat
     4021 aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca
     4081 gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca
     4141 tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg
     4201 cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat
     4261 gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga
     4321 tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg
     4381 agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat
     4441 tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca
     4501 gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc
     4561 ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg
     4621 ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca
     4681 acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca
     4741 tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc
     4801 gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt
     4861 tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg
     4921 ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt
     4981 aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca
     5041 tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg
     5101 aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg
     5161 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg
     5221 tagaggatcg agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag
     5281 cggataacaa ttcccctcta gaaataattt tgtttaactt taagaaggag atatactatg
     5341 gctagcagcc atcatcatca tcatcatagc agcggcctgg tgccgcgcgg cagctccatg
     5401 ggtaacagcg agctggcggc gcgtatcctg atcattctgt tccagcaact ggtggagctg
     5461 gcgcgtctgg cgattgaaag cggcgacgag gaactgctgc gtcgtgtgag cgagtggctg
     5521 gaggaagtta tcaaggatat gcgtcgtgtg gttgaacagg cgctgcgtga gggtaacagc
     5581 gaactggcgg ctcgtatcct gattattctg tttcagcaac tggttgagct ggcgcgtctg
     5641 gcgatagaga gcggtgatga agagttatta cgtcgtgtga gcgaatggct ggaggaagtt
     5701 attaaagaca tgcgtcgtgt tgtggagcag gcgctgcgcg aaggtaacag cgaactggcg
     5761 gcacgtatcc tgataattct gtttcaacaa ctggtggagc tggcgcgtct ggcgatcgag
     5821 agcggtgatg aagaattatt acgtcgtgtg agcgagtggc tggaggaagt tataaaagac
     5881 atgcgtcgtg tggtggagca ggcgctgcgt gaaggtaaca gcgaactggc ggcgcgtatc
     5941 ctgataattc tgttccaaca actggttgag ctggcgcgtc tggcgataga aagcggtgat
     6001 gaagagcttt tacgtcgtgt gagcgagtgg ctggaggaag ttataaagga catgcgtcgt
     6061 gttgttgagc aggcgctgcg cgagggtaac agcgaactgg cggcccgtat cctgataatt
     6121 ctgttccagc aactggtgga gctggcgcgt ctggcgatag aatctggtga tgaagagtta
     6181 cttcgtcgtg tgagcgagtg gctggaggaa gttataaagg atatgcgtcg tgttgtggaa
     6241 caggcgctgc gcgagggtaa cagcgaactg gcggcacgta tcctgatcat tctgtttcag
     6301 caactggttg agctggcgcg tctggcgata gaaagtggtg atgaagagct tcttcgtcgt
     6361 gtgagcgagt ggctggagga agttataaag gatatgcgtc gtgtagttga gcaggcgctg
     6421 cgtgagggct aagcggccgc actcgaggat ccggctgcta acaaagcccg aaaggaagct
     6481 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     6541 gtcttgaggg gttttttgct gaaaggagga actatatccg gatatcccgc aagaggcccg
     6601 gcagtaccgg cataaccaag cctatgccta cagcatccag ggtgacggtg ccgaggatga
     6661 cgatgagcgc attgttagat ttcatacacg gtgcctgact gcgttagcaa tttaactgtg
     6721 ataaactacc gcattaaagc ttatc
//
