LOCUS       Exported                3418 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  construction of TCR.
ACCESSION   .
VERSION     .
KEYWORDS    TRBV4-2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3418)
  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 3418)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3418
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     179..201
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     regulatory      262..271
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     complement(1143..1159)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1143..1159)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1156..1178)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1167..1183
                     /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(1191..1221)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1236..1257
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1374..1391)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1545..2133)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1625..1644)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2311..3120)
                     /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     3034..3053
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3112..3131
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(3262..3284)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3384..3403
                     /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 ggctgcaggc tgctctgctg tgcggttctc
      301 tgtctcctgg gagcggtccc catggaaacg ggagttacgc agacaccaag acacctggtc
      361 atgggaatga caaataagaa gtctttgaaa tgtgaacaac atctggggca taacgctatg
      421 tattggtaca agcaaagtgc taagaagcca ctggagctca tgtttgtcta caactttaaa
      481 gaacagactg aaaacaacag tgtgccaagt cgcttctcac ctgaatgccc caacagctct
      541 cacttattcc ttcacctaca caccctgcag ccagaagact cggccctgta tgagacctaa
      601 ggtggtggtg gttctggttc tggttctggt ggtggttctg gtggtggtct cttgaccctg
      661 ccgtgtacca gctgagagac tctaaatcca gtgacaagtc tgtctgccta ttcaccgatt
      721 ttgattctca aacaaatgtg tcacaaagta aggattctga tgtgtatatc acagacaaaa
      781 ctgtgctaga catgaggtct atggacttca agagcaacag tgctgtggcc tggagcaaca
      841 aatctgactt tgcatgtgca aacgccttca acaacagcat tattccagaa gacaccttct
      901 tccccagccc agaaagttcc tgtgatgtca agctggtcga gaaaagcttt gaaacagata
      961 cgaacctaaa ctttcaaaac ctgtcagtga ttgggttccg aatcctcctc ctgaaagtgg
     1021 ccgggtttaa tctgctcatg acgctgcggc tgtggtccag ctaaagcggc cgcgtcgaca
     1081 atcaaatcgg atcccgggcc cgtcgactgc agaggcctgc atgcaagctt ggtgtaatca
     1141 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     1201 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     1261 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     1321 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     1381 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     1441 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     1501 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     1561 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     1621 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     1681 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     1741 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     1801 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     1861 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     1921 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     1981 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2041 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2101 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2161 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2221 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaag cccaatctga
     2281 ataatgttac aaccaattaa ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa
     2341 ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa
     2401 tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc
     2461 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt
     2521 atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg
     2581 catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc
     2641 atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct
     2701 gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc
     2761 atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg ctgtttttcc
     2821 ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt
     2881 cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt
     2941 ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa
     3001 gcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa
     3061 atcagcatcc atgttggaat ttaatcgcgg cctcgacgtt tcccgttgaa tatggctcat
     3121 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt
     3181 tttatcttgt gcaatgtaac atcagagatt ttgagacacg ggccagagct gcatcgcgcg
     3241 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     3301 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     3361 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccat
//
