LOCUS       sequence-244998-        3427 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 3427)
  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 3427)
  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: TRBV7-4
FEATURES             Location/Qualifiers
     source          1..3427
                     /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"
     regulatory      447..456
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1337..1353)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1337..1353)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1350..1372)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1361..1377
                     /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(1385..1391,1392..1409,1410..1415))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1430..1451
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1568..1585)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1739..2327)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1819..1838)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2505..3314)
                     /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     3228..3247
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3306..3325
                     /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 ccatgggcac caggctcctc tgctgggtgg
      481 tcctgggttt cctagggaca gatcacacag gtgctggagt ctcccagtcc ccaaggtaca
      541 aagtcgcaaa gaggggacgg gatgtagctc tcaggtgtga ttcaatttcg ggtcatgtaa
      601 ccctttattg gtaccgacag accctggggc agggctcaga ggttctgact tactcccaga
      661 gtgatgctca acgagacaaa tcagggcggc ccagtggtcg gttctctgca gagaggcctg
      721 agagatccgt ctccactctg aagatccagc gcacagagca gggggactca gctgtgtatg
      781 agacctaagg tggtggtggt tctggttctg gttctggtgg tggttctggt ggtggtctct
      841 gaaccctgac cctgccgtgt accagctgag agactctaaa tccagtgaca agtctgtctg
      901 cctattcacc gattttgatt ctcaaacaaa tgtgtcacaa agtaaggatt ctgatgtgta
      961 tatcacagac aaaactgtgc tagacatgag gtctatggac ttcaagagca acagtgctgt
     1021 ggcctggagc aacaaatctg actttgcatg tgcaaacgcc ttcaacaaca gcattattcc
     1081 agaagacacc ttcttcccca gcccagaaag ttcctgtgat gtcaagctgg tcgagaaaag
     1141 ctttgaaaca gatacgaacc taaactttca aaacctgtca gtgattgggt tccgaatcct
     1201 cctcctgaaa gtggccgggt ttaatctgct catgacgctg cggctgtggt ccagctaaag
     1261 cggccgcgtc gacaatcaaa tcggatcccg ggcccgtcga ctgcagaggc ctgcatgcaa
     1321 gcttggtgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     1381 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     1441 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     1501 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     1561 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     1621 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1681 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1741 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1801 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1861 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1921 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     1981 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2041 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2101 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2161 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2221 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2281 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2341 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2401 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2461 caagcccaat ctgaataatg ttacaaccaa ttaaccaatt ctgattagaa aaactcatcg
     2521 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa
     2581 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc
     2641 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg
     2701 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
     2761 ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
     2821 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga
     2881 aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg
     2941 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg
     3001 aatgctgttt ttccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata
     3061 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca
     3121 tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg
     3181 ggcttcccat acaagcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat
     3241 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctcga cgtttcccgt
     3301 tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt
     3361 catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga cacgggccag
     3421 agctgca
//
