LOCUS       sequence-244229-        3424 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 3424)
  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 3424)
  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: TRBV5-6
FEATURES             Location/Qualifiers
     source          1..3424
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     179..201
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     regulatory      262..271
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(580..591)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1149..1165)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1149..1165)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1162..1184)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1173..1189
                     /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(1197..1203,1204..1221,1222..1227))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1242..1263
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1380..1397)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1551..2139)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1631..1650)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2317..3126)
                     /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     3040..3059
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3118..3137
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(3268..3290)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3390..3409
                     /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 ggccccgggc tcctctgctg ggcactgctt
      301 tgtctcctgg gagcaggctt agtggacgct ggagtcaccc aaagtcccac acacctgatc
      361 aaaacgagag gacagcaagt gactctgaga tgctctccta agtctgggca tgacactgtg
      421 tcctggtacc aacaggccct gggtcagggg ccccagttta tctttcagta ttatgaggag
      481 gaagagagac agagaggcaa cttccctgat cgattctcag gtcaccagtt ccctaactat
      541 agctctgagc tgaatgtgaa cgccttgttg ctgggggact cggccctcta tgagacctaa
      601 ggtggtggtg gttctggttc tggttctggt ggtggttctg gtggtggtct ctgaaccctg
      661 accctgccgt gtaccagctg agagactcta aatccagtga caagtctgtc tgcctattca
      721 ccgattttga ttctcaaaca aatgtgtcac aaagtaagga ttctgatgtg tatatcacag
      781 acaaaactgt gctagacatg aggtctatgg acttcaagag caacagtgct gtggcctgga
      841 gcaacaaatc tgactttgca tgtgcaaacg ccttcaacaa cagcattatt ccagaagaca
      901 ccttcttccc cagcccagaa agttcctgtg atgtcaagct ggtcgagaaa agctttgaaa
      961 cagatacgaa cctaaacttt caaaacctgt cagtgattgg gttccgaatc ctcctcctga
     1021 aagtggccgg gtttaatctg ctcatgacgc tgcggctgtg gtccagctaa agcggccgcg
     1081 tcgacaatca aatcggatcc cgggcccgtc gactgcagag gcctgcatgc aagcttggtg
     1141 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     1201 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     1261 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     1321 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     1381 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     1441 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     1501 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1561 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     1621 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     1681 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     1741 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     1801 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     1861 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     1921 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     1981 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2041 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2101 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2161 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     2221 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaagccca
     2281 atctgaataa tgttacaacc aattaaccaa ttctgattag aaaaactcat cgagcatcaa
     2341 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     2401 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     2461 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     2521 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     2581 tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     2641 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg
     2701 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     2761 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     2821 ttttccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     2881 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     2941 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     3001 atacaagcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     3061 atataaatca gcatccatgt tggaatttaa tcgcggcctc gacgtttccc gttgaatatg
     3121 gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga
     3181 tatattttta tcttgtgcaa tgtaacatca gagattttga gacacgggcc agagctgcat
     3241 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     3301 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     3361 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     3421 ccat
//
