LOCUS       sequence-343820-        7258 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  de novo thick circular tandem repeat proteins with 9 repeats.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7258)
  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 7258)
  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: tcTRP9
FEATURES             Location/Qualifiers
     source          1..7258
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..47)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        115..219
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(220..288,289..1080)
                     /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(438..457)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1251..1839
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1740..1759
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1993..2010
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2025..2164
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2090..2109)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2250..2272
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2266..2457)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4812..4831
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4851..4928)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5134..5153
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5237..5256
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5237..5255
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5256..5280
                     /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             5295..5317
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5336..5353
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5363..5380
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(6996..7014)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7010..7057
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(7232..7237,7238..7254,7255..7258,1..2))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 aaacatgaga attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
       61 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
      121 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
      181 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
      241 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
      301 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
      361 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
      421 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
      481 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
      541 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
      601 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
      661 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
      721 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg
      781 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
      841 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
      901 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
      961 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     1021 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     1081 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     1141 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     1201 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     1261 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     1321 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     1381 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     1441 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     1501 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     1561 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     1621 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     1681 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     1741 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     1801 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     1861 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     1921 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     1981 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     2041 ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct
     2101 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
     2161 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     2221 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     2281 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga
     2341 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa
     2401 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga
     2461 tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag
     2521 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg
     2581 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca
     2641 gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca
     2701 gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg
     2761 gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct
     2821 tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc
     2881 tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct
     2941 gcccgagatg cgccgcgtgc ggctgctgga gatggcggac gcgatggata tgttctgcca
     3001 agggttggtt tgcgcattca cagttctccg caagaattga ttggctccaa ttcttggagt
     3061 ggtgaatccg ttagcgaggt gccgccggct tccattcagg tcgaggtggc ccggctccat
     3121 gcaccgcgac gcaacgcggg gaggcagaca aggtataggg cggcgcctac aatccatgcc
     3181 aacccgttcc atgtgctcgc cgaggcggca taaatcgccg tgacgatcag cggtccaatg
     3241 atcgaagtta ggctggtaag agccgcgagc gatccttgaa gctgtccctg atggtcgtca
     3301 tctacctgcc tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga
     3361 agaatcataa tggggaaggc catccagcct cgcgtcgcga acgccagcaa gacgtagccc
     3421 agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg
     3481 ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg
     3541 ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc
     3601 ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc
     3661 atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga
     3721 gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
     3781 ttccagtcgg gaaactgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3841 gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc
     3901 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc
     3961 cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct
     4021 tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta
     4081 atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg
     4141 atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct
     4201 tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga
     4261 cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc
     4321 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg
     4381 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct
     4441 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt
     4501 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc
     4561 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc
     4621 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc
     4681 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact
     4741 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga
     4801 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc
     4861 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg
     4921 atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag
     4981 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc
     5041 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
     5101 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
     5161 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat
     5221 ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc
     5281 cctctagaaa taattttgtt taactttaag aaggagatat actatggcta gcagccatca
     5341 tcatcatcat catagcagcg gcctggtgcc gcgcggcagc tccatgggca acctggaact
     5401 ggcactgaaa gctctgcaga ttctggtgaa tgcggcctat gttctggcgg aaatcgcccg
     5461 tgatcgcggt aacgaagaac tgctggaaaa agcggctcgt ctggccgaag aagcggcacg
     5521 tcaggcagaa gaaattgcac gtcaagctcg caaagaaggc aacctggaac tggcgctgaa
     5581 agccctgcaa attctggtca atgcagctta cgtgctggca gaaatcgctc gtgaccgcgg
     5641 taatgaggag ctgctggaaa aagcggcccg tctggcagaa gaagcagctc gccaggccga
     5701 agaaatcgcg cgtcaagccc gtaaagaagg caatctggaa ctggccctga aagccctgca
     5761 aatcctggtt aatgcggcct atgtcctggc cgaaattgcg cgcgaccgtg gcaatgagga
     5821 attactggaa aaagcggcgc gcctggccga agaagcggcc cgccaagccg aagaaatcgc
     5881 ccgccaggcc cgtaaagaag gcaacttaga gctggctctg aaagcgctgc aaatcctggt
     5941 gaatgcagct tacgttctgg ccgaaattgc ccgcgaccgc ggtaatgagg aacttctgga
     6001 aaaagccgca cgtctggccg aagaagcagc tcgtcaagcg gaagaaattg cccgtcaagc
     6061 ccgcaaagaa ggcaatttag agctggccct gaaagcgctg cagattctgg tcaatgcggc
     6121 ctatgtgctg gccgaaattg cgcgtgaccg tggcaatgag gaactcctgg aaaaagcggc
     6181 gcgtttagcc gaagaagcgg cccgtcaagc cgaagaaatc gctcgccagg cccgaaaaga
     6241 aggcaattta gaattagctt taaaagcgct gcaaattctg gtgaacgccg cttacgtcct
     6301 ggccgaaatt gcccgtgatc gcggtaatga ggaactactg gaaaaagccg cccgcctggc
     6361 cgaagaagca gcccgtcagg cggaagaaat tgctcgccag gcccggaaag aaggcaattt
     6421 ggagctggcc ctgaaagcac tgcaaatcct ggtcaacgcc gcgtacgtgc tggccgaaat
     6481 cgcgcgcgac cgtggcaacg aggagttatt agaaaaagcg gcgcgtcttg ccgaagaagc
     6541 ggcgcgccag gccgaagaaa ttgcgcgcca ggccagaaaa gaaggcaatc ttgagctggc
     6601 tctgaaagct ctgcaaatcc tggtcaatgc cgcctacgtt ctggcagaaa tcgcgcgtga
     6661 ccgcggtaat gaagagttac tggaaaaagc cgcgcgctta gccgaagaag cagcgcgcca
     6721 agccgaagaa attgcgcgtc aagcacgtaa agaaggcaac cttgagctgg cactgaaggc
     6781 gttacaaatt ctggttaacg cggcctatgt gctggcggaa atcgcgcgcg accgcggcaa
     6841 tgaggaactg ttagaaaaag cggcacgctt agccgaagaa gcggcgcgtc aagccgaaga
     6901 aattgctcgc caagcacgca aagaaggcta agcggccgca ctcgaggatc cggctgctaa
     6961 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
     7021 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
     7081 atatcccgca agaggcccgg cagtaccggc ataaccaagc ctatgcctac agcatccagg
     7141 gtgacggtgc cgaggatgac gatgagcgca ttgttagatt tcatacacgg tgcctgactg
     7201 cgttagcaat ttaactgtga taaactaccg cattaaagct tatcgatgat aagctgtc
//
