LOCUS       sequence-470000-        3403 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  backbone for makeTCR assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3403)
  TITLE     makeTCR collection
REFERENCE   2  (bases 1 to 3403)
  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"
COMMENT     Sequence Label: makeTCR_HDR_Hs.TRAC_Backbone
FEATURES             Location/Qualifiers
     source          1..3403
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    246..453
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(840..856)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(840..856)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(853..875)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1028..1616)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1108..1127)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1797..2606)
                     /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     2520..2539
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2598..2617
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(2748..2770)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2955..2977
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2969..2986
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2970..2986
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(3352..3403,1..2)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
ORIGIN
        1 ccatggagac gtgctaaccg tatcgcattg gccataccga agatggccgc gaagagaact
       61 agcccgggac catttaaatt gagccacgtc tcataaccaa gcacgcagca atgcagctca
      121 aaacgcttag cctagccaca cccccacggg aaacagcagt gattaacctt tagcaataaa
      181 cgaaagttta actaagctat actaacccca gggttggtca atttcgtgcc agccacactt
      241 aactactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
      301 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
      361 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
      421 ggattgggaa gagaatagca ggcatgctgg ggataccagc tgagagactc taaatccagt
      481 gacaagtctg tctgcctatt caccgatttt gattctcaaa caaatgtgtc acaaagtaag
      541 gattctgatg tgtatatcac agacaaaact gtgctagaca tgaggtctat ggacttcaag
      601 agcaacagtg ctgtggcctg gagcaacaaa tctgactttg catgtgcaaa cgccttcaac
      661 aacagcatta ttccagaaga caccttcttc cccagcccag gtaagggcag ctttggtgcc
      721 ttcgcaggct gtttccttgc ttcaggaatg gccaggttct gcccagagct ctggtcaatg
      781 atgaggctag gtggaggctc agtgatgata agtctgcgat ggtggatgca tgtgtcatgg
      841 tcatagctgt ttcctgtgtg aaattgttat ccgctcagag ggcacaatcc tattccgcgc
      901 tatccgacaa tctccaagac attaggtgga gttcagttcg gcgtatggca tatgtcgctg
      961 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     1021 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     1081 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     1141 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     1201 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     1261 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     1321 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     1381 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     1441 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     1501 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     1561 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     1621 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     1681 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     1741 gaagttttaa atcaagccca atctgaataa tgttacaacc aattaaccaa ttctgattag
     1801 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca
     1861 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg
     1921 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt
     1981 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa
     2041 tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca
     2101 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc
     2161 tgagcaagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc
     2221 aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct
     2281 tctaatacct ggaatgctgt ttttccgggg atcgcagtgg tgagtaacca tgcatcatca
     2341 ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt
     2401 ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac
     2461 tctggcgcat cgggcttccc atacaagcga tagattgtcg cacctgattg cccgacatta
     2521 tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctc
     2581 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
     2641 agttttattg ttcatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga
     2701 gacacgggcc agagctgcat cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     2761 cagctcccgg gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     2821 agggcgcgtc agcgggtgtt ggctgttgtt tgtcgcatta tacgagcgtc caggttggga
     2881 tacctgaaac aaaacccatc gtacggccaa ggaagtctcc aataactgtg atccaccaca
     2941 agcgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtcatg cataatccgc
     3001 acgcatctgg aataaggaag tgccattccg cctgaccttt tcaggtttcc ttgagtggca
     3061 ggccaggcct ggccgtgaac gttcactgaa atcatggcct cttggccaag attgatagct
     3121 tgtgcctgtc cctgagtccc agtccatcac gagcagctgg tttctaagat gctatttccc
     3181 gtataaagca tagaccgtga cttgccagcc ccacagagcc ccgcccttgt ccatcactgg
     3241 catctggact ccagcctggg ttggggcaaa gagggaaatg agatcatgtc ctaaccctga
     3301 tcctcttgtc ccacagatat ccagaaccct gaccctgcct ccggatccgg agagggcagg
     3361 ggatctctcc ttacttgtgg cgacgtggag gagaaccccg gcc
//
