LOCUS       sequence-343877-        6001 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human Nck2 SH2 domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6001)
  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 6001)
  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: SH2-free
FEATURES             Location/Qualifiers
     source          1..6001
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        63..167
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(168..236,237..1028)
                     /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(386..405)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1199..1787
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1688..1707
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1941..1958
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1973..2112
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2038..2057)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2198..2220
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2214..2405)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4760..4779
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4799..4876)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5082..5101
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5185..5204
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5185..5203
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5204..5228
                     /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             5243..5265
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5284..5301
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5311..5328
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(5687..5705)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5701..5748
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(5923..5928,5929..5945,5946..5951))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(5978..5996)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
       61 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
      121 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
      181 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
      241 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
      301 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
      361 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
      421 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
      481 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
      541 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
      601 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
      661 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg
      721 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
      781 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
      841 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
      901 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
      961 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     1021 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     1081 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     1141 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     1201 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     1261 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     1321 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     1381 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     1441 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     1501 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     1561 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     1621 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     1681 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     1741 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     1801 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     1861 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     1921 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     1981 ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac
     2041 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
     2101 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     2161 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     2221 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
     2281 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
     2341 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
     2401 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
     2461 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
     2521 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
     2581 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
     2641 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac
     2701 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag
     2761 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc
     2821 cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg tggccaggac
     2881 ccaacgctgc ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg
     2941 ttctgccaag ggttggtttg cgcattcaca gttctccgca agaattgatt ggctccaatt
     3001 cttggagtgg tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc gaggtggccc
     3061 ggctccatgc accgcgacgc aacgcgggga ggcagacaag gtatagggcg gcgcctacaa
     3121 tccatgccaa cccgttccat gtgctcgccg aggcggcata aatcgccgtg acgatcagcg
     3181 gtccaatgat cgaagttagg ctggtaagag ccgcgagcga tccttgaagc tgtccctgat
     3241 ggtcgtcatc tacctgcctg gacagcatgg cctgcaacgc gggcatcccg atgccgccgg
     3301 aagcgagaag aatcataatg gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga
     3361 cgtagcccag cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt
     3421 tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa
     3481 gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga
     3541 gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga
     3601 cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca
     3661 tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac
     3721 tgcccgcttt ccagtcggga aactgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3781 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg
     3841 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc
     3901 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg
     3961 agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg
     4021 actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag
     4081 tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc
     4141 agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc
     4201 cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct
     4261 ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa
     4321 taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc
     4381 aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac
     4441 tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt
     4501 ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga
     4561 caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact
     4621 gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg
     4681 cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa
     4741 cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat
     4801 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc
     4861 gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag
     4921 cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag
     4981 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
     5041 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
     5101 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
     5161 atcgagatct cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata
     5221 acaattcccc tctagaaata attttgttta actttaagaa ggagatatac tatggctagc
     5281 agccatcatc atcatcatca tagcagcggc ctggtgccgc gcggcagcca tatgtccgag
     5341 tggtactacg ggaacgtgac gcggcaccag gccgagtgcg ccctcaacga gcggggcgtg
     5401 gagggcgact tcctcattag ggacagcgag tcctcgccca gcgacttctc cgtgtccctt
     5461 aaagcgtcag ggaagaacaa acacttcaag gtgcagctcg tggacaatgt ctactgcatt
     5521 gggcagcggc gcttccacac catggacgag ctggtggaac actacaaaaa ggcgcccatc
     5581 ttcaccagcg agcacgggga gaagctctac ctcgtcaggg ccctgcagtg actcgaggat
     5641 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     5701 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga
     5761 actatatccg gatatcccgc aagaggcccg gcagtaccgg cataaccaag cctatgccta
     5821 cagcatccag ggtgacggtg ccgaggatga cgatgagcgc attgttagat ttcatacacg
     5881 gtgcctgact gcgttagcaa tttaactgtg ataaactacc gcattaaagc ttatcgatga
     5941 taagctgtca aacatgagaa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     6001 t
//
