LOCUS       sequence-244201-        3412 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 3412)
  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 3412)
  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: TRBV30
FEATURES             Location/Qualifiers
     source          1..3412
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     179..201
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1137..1153)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1137..1153)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1150..1172)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1161..1177
                     /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(1185..1191,1192..1209,1210..1215))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1230..1251
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1368..1385)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1539..2127)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1619..1638)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2305..3114)
                     /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     3028..3047
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3106..3125
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(3256..3278)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3378..3397
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg
       61 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
      121 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
      181 cagtcacgac gttgtaaaac gacggccaga gaattcgagc tcggtacctc gcgaatacat
      241 ctagatgggg atccaccggt cgccaccatg ctctgctctc tccttgccct tctcctgggc
      301 actttctttg gggtcagatc tcagactatt catcaatggc cagcgaccct ggtgcagcct
      361 gtgggcagcc cgctctctct ggagtgcact gtggagggaa catcaaaccc caacctatac
      421 tggtaccgac aggctgcagg caggggcctc cagctgctct tctactccgt tggtattggc
      481 cagatcagct ctgaggtgcc ccagaatctc tcagcctcca gaccccagga ccggcagttc
      541 atcctgagtt ctaagaagct ccttctcagt gactctggct tctatgagac ctaaggtggt
      601 ggtggttctg gttctggttc tggtggtggt tctggtggtg gtctcttgac cctgccgtgt
      661 accagctgag agactctaaa tccagtgaca agtctgtctg cctattcacc gattttgatt
      721 ctcaaacaaa tgtgtcacaa agtaaggatt ctgatgtgta tatcacagac aaaactgtgc
      781 tagacatgag gtctatggac ttcaagagca acagtgctgt ggcctggagc aacaaatctg
      841 actttgcatg tgcaaacgcc ttcaacaaca gcattattcc agaagacacc ttcttcccca
      901 gcccagaaag ttcctgtgat gtcaagctgg tcgagaaaag ctttgaaaca gatacgaacc
      961 taaactttca aaacctgtca gtgattgggt tccgaatcct cctcctgaaa gtggccgggt
     1021 ttaatctgct catgacgctg cggctgtggt ccagctaaag cggccgcgtc gacaatcaaa
     1081 tcggatcccg ggcccgtcga ctgcagaggc ctgcatgcaa gcttggtgta atcatggtca
     1141 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     1201 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     1261 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     1321 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     1381 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     1441 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     1501 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     1561 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     1621 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     1681 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     1741 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     1801 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     1861 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     1921 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     1981 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2041 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     2101 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     2161 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     2221 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caagcccaat ctgaataatg
     2281 ttacaaccaa ttaaccaatt ctgattagaa aaactcatcg agcatcaaat gaaactgcaa
     2341 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
     2401 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
     2461 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
     2521 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc
     2581 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac
     2641 caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa
     2701 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac
     2761 aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt ttccggggat
     2821 cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag
     2881 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac
     2941 gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaagcgata
     3001 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc
     3061 atccatgttg gaatttaatc gcggcctcga cgtttcccgt tgaatatggc tcataacacc
     3121 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc
     3181 ttgtgcaatg taacatcaga gattttgaga cacgggccag agctgcatcg cgcgtttcgg
     3241 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     3301 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     3361 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc at
//
