LOCUS       sequence-169546-        4407 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HLA.A2_GFP fusion with 2 glycines in between.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4407)
  AUTHORS   Walchli S, Kumari S, Fallang LE, Sand KM, Yang W, Landsverk OJ, 
            Bakke O, Olweus J, Gregers TF
  TITLE     Invariant chain as a vehicle to load antigenic peptides on human MHC
            class I for cytotoxic T-cell activation.
  JOURNAL   Eur J Immunol. 2014 Mar;44(3):774-84. doi: 10.1002/eji.201343671. 
            Epub 2013 Dec 27.
  PUBMED    24293164
REFERENCE   2  (bases 1 to 4407)
  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: 
            "10.1002/eji.201343671"; journalName: "Eur J Immunol"; date: 
            "2014-03"; volume: "44"; issue: "3"; pages: "774-84"
COMMENT     Sequence Label: pENTR-HLA-A2_GFP
FEATURES             Location/Qualifiers
     source          1..4407
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(1790..1792,1793..1795,1796..2509)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1790..1792,1793..1795,1796..2506)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1835..1856)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2096..2115)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2443..2464
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2532..2631)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2648..2667)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2649..2667)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2672..2688)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2672..2688)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2790..2809)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2801..3610
                     /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     complement(2868..2887)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3757..4345
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4246..4265
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc gcggccgccc ccttcaccat ggccgtcatg gcgccccgaa ccctcgtcct
      721 gctactctcg ggggctctgg ccctgaccca gacctgggcg ggctctcact ccatgaggta
      781 tttcttcaca tccgtgtccc ggcccggccg cggggagccc cgcttcatcg cagtgggcta
      841 cgtggacgac acgcagttcg tgcggttcga cagcgacgcc gcgagccaga ggatggagcc
      901 gcgggcgccg tggatagagc aggagggtcc ggagtattgg gacggggaga cacggaaagt
      961 gaaggcccac tcacagactc accgagtgga cctggggacc ctgcgcggct actacaacca
     1021 gagcgaggcc ggttctcaca ccgtccagag gatgtatggc tgcgacgtgg ggtcggactg
     1081 gcgcttcctc cgcgggtacc accagtacgc ctacgacggc aaggattaca tcgccctgaa
     1141 agaggacctg cgctcttgga ccgcggcgga catggcagct cagaccacca agcacaagtg
     1201 ggaggcggcc catgtggcgg agcagttgag agcctacctg gagggcacgt gcgtggagtg
     1261 gctccgcaga tacctggaga acgggaagga gacgctgcag cgcacggacg cccccaaaac
     1321 gcatatgact caccacgctg tctctgacca tgaagccacc ctgaggtgct gggccctgag
     1381 cttctaccct gcggagatca cactgacctg gcagcgggat ggggaggacc agacccagga
     1441 cacggagctc gtggagacca ggcctgcagg ggatggaacc ttccagaagt gggcggctgt
     1501 ggtggtgcct tctggacagg agcagagata cacctgccat gtgcagcatg agggtttgcc
     1561 caagcccctc accctgagat gggagccgtc ttcccagccc accatcccca tcgtgggcat
     1621 cattgctggc ctggttctct ttggagctgt gatcactgga gctgtggtcg ctgctgtgat
     1681 gtggaggagg aagagctcag atagaaaagg agggagctac tctcaggctg caagcagtga
     1741 cagtgcccag ggctctgatg tgtctctcac agcttgtaaa gtgggaggta tggtgagcaa
     1801 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     1861 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     1921 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1981 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     2041 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     2101 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     2161 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     2221 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     2281 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     2341 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     2401 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     2461 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaac tcgagaaggg
     2521 tgggcgcgcc gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     2581 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     2641 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     2701 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     2761 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     2821 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     2881 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     2941 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     3001 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     3061 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     3121 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     3181 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     3241 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     3301 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     3361 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     3421 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     3481 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     3541 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     3601 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     3661 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     3721 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3781 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3841 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3901 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3961 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4021 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4081 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4141 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4201 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4261 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4321 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4381 cttttgctgg ccttttgctc acatgtt
//
