LOCUS       sequence-235528-        3412 bp    DNA     circular SYN 14-APR-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 Apr 14, 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     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(1322..1338)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1322..1338)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1335..1357)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1346..1362
                     /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(1370..1376,1377..1394,1395..1400))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1415..1436
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1553..1570)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1724..2312)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1804..1823)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2490..3299)
                     /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     3213..3232
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3291..3310
                     /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 ccatgctctg ctctctcctt gcccttctcc
      481 tgggcacttt ctttggggtc agatctcaga ctattcatca atggccagcg accctggtgc
      541 agcctgtggg cagcccgctc tctctggagt gcactgtgga gggaacatca aaccccaacc
      601 tatactggta ccgacaggct gcaggcaggg gcctccagct gctcttctac tccgttggta
      661 ttggccagat cagctctgag gtgccccaga atctctcagc ctccagaccc caggaccggc
      721 agttcatcct gagttctaag aagctccttc tcagtgactc tggcttctat gagacctaag
      781 gtggtggtgg ttctggttct ggttctggtg gtggttctgg tggtggtctc ttgaccctgc
      841 cgtgtaccag ctgagagact ctaaatccag tgacaagtct gtctgcctat tcaccgattt
      901 tgattctcaa acaaatgtgt cacaaagtaa ggattctgat gtgtatatca cagacaaaac
      961 tgtgctagac atgaggtcta tggacttcaa gagcaacagt gctgtggcct ggagcaacaa
     1021 atctgacttt gcatgtgcaa acgccttcaa caacagcatt attccagaag acaccttctt
     1081 ccccagccca gaaagttcct gtgatgtcaa gctggtcgag aaaagctttg aaacagatac
     1141 gaacctaaac tttcaaaacc tgtcagtgat tgggttccga atcctcctcc tgaaagtggc
     1201 cgggtttaat ctgctcatga cgctgcggct gtggtccagc taaagcggcc gcgtcgacaa
     1261 tcaaatcgga tcccgggccc gtcgactgca gaggcctgca tgcaagcttg gtgtaatcat
     1321 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     1381 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     1441 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     1501 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     1561 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     1621 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     1681 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     1741 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     1801 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     1861 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     1921 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     1981 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     2041 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     2101 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     2161 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     2221 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     2281 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     2341 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     2401 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaagc ccaatctgaa
     2461 taatgttaca accaattaac caattctgat tagaaaaact catcgagcat caaatgaaac
     2521 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     2581 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     2641 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     2701 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc
     2761 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     2821 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg
     2881 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
     2941 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgtttttccg
     3001 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     3061 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     3121 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaag
     3181 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     3241 tcagcatcca tgttggaatt taatcgcggc ctcgacgttt cccgttgaat atggctcata
     3301 acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga tgatatattt
     3361 ttatcttgtg caatgtaaca tcagagattt tgagacacgg gccagagctg ca
//
