LOCUS       sequence-343875-        7876 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  de novo thick circular tandem repeat protein trimer formed from 
            8-repeat monomers containing 2 copies of human Nck2 SH2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7876)
  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 7876)
  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_SH2sub6
FEATURES             Location/Qualifiers
     source          1..7876
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(26..44)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        112..216
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(217..285,286..1077)
                     /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(435..454)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1248..1836
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1737..1756
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1990..2007
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2022..2161
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2087..2106)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2247..2269
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2263..2454)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4809..4828
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4848..4925)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5131..5150
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5234..5253
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5234..5252
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5253..5277
                     /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             5292..5314
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5333..5350
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5360..5377
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(7611..7629)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7625..7672
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(7847..7852,7853..7869,7870..7875))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 catgagaatt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
       61 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
      121 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
      181 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
      241 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
      301 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
      361 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
      421 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
      481 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
      541 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
      601 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
      661 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
      721 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
      781 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
      841 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
      901 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
      961 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     1021 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     1081 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     1141 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     1201 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     1261 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     1321 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     1381 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     1441 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     1501 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     1561 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     1621 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     1681 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     1741 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     1801 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     1861 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     1921 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     1981 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     2041 cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg
     2101 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     2161 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     2221 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     2281 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc
     2341 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     2401 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc
     2461 ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat
     2521 gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa
     2581 acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg
     2641 cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat
     2701 ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa
     2761 accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca
     2821 cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag
     2881 ccgggtcctc aacgacagga gcacgatcat gcgcacccgt ggccaggacc caacgctgcc
     2941 cgagatgcgc cgcgtgcggc tgctggagat ggcggacgcg atggatatgt tctgccaagg
     3001 gttggtttgc gcattcacag ttctccgcaa gaattgattg gctccaattc ttggagtggt
     3061 gaatccgtta gcgaggtgcc gccggcttcc attcaggtcg aggtggcccg gctccatgca
     3121 ccgcgacgca acgcggggag gcagacaagg tatagggcgg cgcctacaat ccatgccaac
     3181 ccgttccatg tgctcgccga ggcggcataa atcgccgtga cgatcagcgg tccaatgatc
     3241 gaagttaggc tggtaagagc cgcgagcgat ccttgaagct gtccctgatg gtcgtcatct
     3301 acctgcctgg acagcatggc ctgcaacgcg ggcatcccga tgccgccgga agcgagaaga
     3361 atcataatgg ggaaggccat ccagcctcgc gtcgcgaacg ccagcaagac gtagcccagc
     3421 gcgtcggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga
     3481 ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg
     3541 atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc
     3601 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg
     3661 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
     3721 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
     3781 cagtcgggaa actgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3841 gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga
     3901 ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc
     3961 agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg
     4021 gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg
     4081 gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg
     4141 ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc
     4201 cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc
     4261 agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat
     4321 gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg
     4381 atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc
     4441 acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc
     4501 gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac
     4561 accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac
     4621 ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc
     4681 agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt
     4741 tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa
     4801 gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg
     4861 aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg
     4921 gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag
     4981 taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc
     5041 caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag
     5101 cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac
     5161 cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc
     5221 gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa caattcccct
     5281 ctagaaataa ttttgtttaa ctttaagaag gagatatact atggctagca gccatcatca
     5341 tcatcatcat agcagcggcc tggtgccgcg cggcagctcc atgggcaaca gcgagctggc
     5401 ggcgcgtatc ctgatcattc tgttccagca actggtggag ctggcgcgtc tggcgattga
     5461 aagcggtgac gaggaactgc tgcgtcgtgt gagcgagtgg ctggaggaag ttatcaagga
     5521 tatgcgtcgt gtggttgaac aggcgctgcg tgagggtaac agcgaactgg ctgctcgtat
     5581 cctgattatc ctgtttcagc aactggttga gctggcgcgc ctggcgattg aaagcggcga
     5641 tgaagaactg ttacgtcgtg tgagcgaatg gctggaggaa gttattaaag acatgcgtcg
     5701 tgttgttgag caggcgctgc gtgaaggtgg cggtagcgac ggtcaggata agggtagcga
     5761 cggccaagat aaaggcggta gcgagtggta ctatggcaac gtgacccgtc accaagcgga
     5821 atgcgcgctg aacgagcgtg gtgttgaagg cgacttcctg atccgtgata gcgaaagcag
     5881 cccgagcgac tttagcgtga gcctgaaggc gagcggcaag aacaaacact tcaaagtgca
     5941 gctggttgac aacgtgtact gcatcggcca acgtcgtttt cacacgatgg acgagctggt
     6001 tgagcactac aagaaagcgc cgattttcac cagcgagcac ggtgaaaagc tgtatctggt
     6061 tcgtgcgctg cagggcggta gcgacggtca agacaaaggt agcgacggcc aggataaagg
     6121 caacagcgag ctggctgctc gtatcctgat aatcctgttt caacaactgg tggagctggc
     6181 tcgcctggcg atcgagagcg gcgatgagga actgttacgt cgtgtgagcg agtggctgga
     6241 ggaagttata aaagacatgc gtcgtgtggt ggagcaagcg ctgcgcgaag gtaacagcga
     6301 actggccgct cgtatcctga ttattctgtt tcaacaactg gttgagctgg ctcgtctggc
     6361 gatcgagagc ggtgatgaag aactgcttcg tcgtgtgagc gagtggctgg aggaagttat
     6421 aaaggacatg cgtcgtgttg tggagcaggc gctgcgcgag ggtaacagcg aactggcagc
     6481 gcgtatcctg attatcttat ttcagcaact ggtggagctg gcacgcctgg cgatcgaatc
     6541 tggtgatgaa gaactgctcc gtcgtgtgag cgagtggctg gaggaagtta taaaggatat
     6601 gcgtcgtgtt gtggaacagg cgctgcgcga gggtaacagc gaactggcgg ctcgtatcct
     6661 gatcatcctg ttccaacaac tggttgagct ggcacgcctg gcgattgaga gcggcgatga
     6721 ggaactgctt cgtcgtgtga gcgagtggct ggaggaagtt ataaaggata tgcgtcgtgt
     6781 agtggaacag gcgctgcgtg aaggcggtgg cagcgatggc caagacaagg gtagcgatgg
     6841 tcaagataaa ggtggcagcg aatggtatta cggtaatgtt acccgccatc aggcggagtg
     6901 cgcgctgaat gaacgtggtg ttgagggtga cttcctgatt cgcgactctg aaagcagccc
     6961 tagcgacttt agcgttagcc tgaaggcgag cggcaaaaac aaacacttta aagttcagct
     7021 ggttgacaac gtttattgca ttggtcaacg tcgtttccat acgatggacg agctggttga
     7081 gcattataag aaagcgccga tctttaccag cgaacatggt gaaaaactgt acctggtgcg
     7141 tgcgctgcag ggtggcagcg acggtcaaga caagggtagc gacggtcaag ataagggcaa
     7201 cagcgagctg gccgctcgta tcctgataat tctgtttcag caactggtgg agctggcccg
     7261 attagcgatc gagagcggcg acgaagaact gctacgtcgt gtgagcgagt ggctggagga
     7321 agttataaag gatatgcgtc gtgtcgttga acaagcgctg cgagaaggta acagcgaact
     7381 ggcagcacgt atcctgataa tattattcca gcaactggtt gagctggccc gtttagctat
     7441 tgaaagcggc gacgaggaac tgttacgtcg tgtgagcgag tggctggagg aagttataaa
     7501 ggatatgcgt cgtgtagtag aacaggcgct gcgcgagggt taataagcgg ccgcactcga
     7561 ggatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     7621 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     7681 aggaactata tccggatatc ccgcaagagg cccggcagta ccggcataac caagcctatg
     7741 cctacagcat ccagggtgac ggtgccgagg atgacgatga gcgcattgtt agatttcata
     7801 cacggtgcct gactgcgtta gcaatttaac tgtgataaac taccgcatta aagcttatcg
     7861 atgataagct gtcaaa
//
