LOCUS       sequence-245037-        3415 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  construction of TCR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3415)
  AUTHORS   Hu Z, Anandappa AJ, Sun J, Kim J, Leet DE, Bozym DJ, Chen C, 
            Williams L, Shukla SA, Zhang W, Tabbaa D, Steelman S, Olive O, Livak
            KJ, Kishi H, Muraguchi A, Guleria I, Stevens J, Lane WJ, Burkhardt 
            UE, Fritsch EF, Neuberg D, Ott PA, Keskin DB, Hacohen N, Wu CJ
  TITLE     A cloning and expression system to probe T-cell receptor specificity
            and assess functional avidity to neoantigens.
  JOURNAL   Blood. 2018 Nov 1;132(18):1911-1921. doi: 
            10.1182/blood-2018-04-843763. Epub 2018 Aug 27.
  PUBMED    30150207
REFERENCE   2  (bases 1 to 3415)
  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"; doi: 
            "10.1182/blood-2018-04-843763"; journalName: "Blood"; date: 
            "2018-11-1- 1"; volume: "132"; issue: "18"; pages: "1911-1921"
COMMENT     Sequence Label: TRBV20-1
FEATURES             Location/Qualifiers
     source          1..3415
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1325..1341)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1325..1341)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1338..1360)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1349..1365
                     /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)."
     promoter        complement(join(1373..1379,1380..1397,1398..1403))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1418..1439
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1556..1573)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1727..2315)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1807..1826)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2493..3302)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3216..3235
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3294..3313
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagagaatt cgagctcggt acctcgcgaa
      421 tacatctaga tggggatcca ccggtcgcca ccatgctgct gcttctgctg cttctggggc
      481 cagcaggctc cgggcttggt gctgtcgtct ctcaacatcc gagctgggtt atctgtaaga
      541 gtggaacctc tgtgaagatc gagtgccgtt ccctggactt tcaggccaca actatgtttt
      601 ggtatcgtca gttcccgaaa cagagtctca tgctgatggc aacttccaat gagggctcca
      661 aggccacata cgagcaaggc gtcgagaagg acaagtttct catcaaccat gcaagcctga
      721 ccttgtccac tctgacagtg accagtgccc atcctgaaga cagcagcttc tacgagacct
      781 aaggtggtgg tggttctggt tctggttctg gtggtggttc tggtggtggt ctcttgaccc
      841 tgccgtgtac cagctgagag actctaaatc cagtgacaag tctgtctgcc tattcaccga
      901 ttttgattct caaacaaatg tgtcacaaag taaggattct gatgtgtata tcacagacaa
      961 aactgtgcta gacatgaggt ctatggactt caagagcaac agtgctgtgg cctggagcaa
     1021 caaatctgac tttgcatgtg caaacgcctt caacaacagc attattccag aagacacctt
     1081 cttccccagc ccagaaagtt cctgtgatgt caagctggtc gagaaaagct ttgaaacaga
     1141 tacgaaccta aactttcaaa acctgtcagt gattgggttc cgaatcctcc tcctgaaagt
     1201 ggccgggttt aatctgctca tgacgctgcg gctgtggtcc agctaaagcg gccgcgtcga
     1261 caatcaaatc ggatcccggg cccgtcgact gcagaggcct gcatgcaagc ttggtgtaat
     1321 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     1381 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     1441 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     1501 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     1561 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     1621 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     1681 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1741 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     1801 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     1861 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     1921 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     1981 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2041 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2101 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2161 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2221 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2281 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2341 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     2401 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca agcccaatct
     2461 gaataatgtt acaaccaatt aaccaattct gattagaaaa actcatcgag catcaaatga
     2521 aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt
     2581 aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct
     2641 gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg
     2701 ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta
     2761 tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc
     2821 gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg
     2881 ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc
     2941 gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttt
     3001 ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg
     3061 gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca
     3121 ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac
     3181 aagcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat
     3241 aaatcagcat ccatgttgga atttaatcgc ggcctcgacg tttcccgttg aatatggctc
     3301 ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca tgatgatata
     3361 tttttatctt gtgcaatgta acatcagaga ttttgagaca cgggccagag ctgca
//
