LOCUS       sequence-350753-        7114 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pAR100 allows cloning of TCR-alpha and TCR-beta genes using 
            restriction enzymes (AflII, ApaI, and SacII)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7114)
  AUTHORS   Sooda A, Rwandamuriye F, Wanjalla CN, Jing L, Koelle DM, Peters B, 
            Leary S, Chopra A, Calderwood MA, Mallal SA, Pavlos R, Watson M, 
            Phillips EJ, Redwood AJ
  TITLE     Abacavir inhibits but does not cause self-reactivity to 
            HLA-B*57:01-restricted EBV specific T cell receptors.
  JOURNAL   Commun Biol. 2022 Feb 16;5(1):133. doi: 10.1038/s42003-022-03058-9.
  PUBMED    35173258
REFERENCE   2  (bases 1 to 7114)
  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"; journalName: "Commun 
            Biol."; date: "2022-02-16"; volume: "5(1):133. doi"; pages: " 
            10.1038/s42003-022-03058-9"
COMMENT     Sequence Label: pAR100
FEATURES             Location/Qualifiers
     source          1..7114
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    6..400
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     CDS             complement(961..1677)
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQRNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      1674..1683
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     LTR             1687..1955
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     primer_bind     complement(1964..1988)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(1985..2187)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2014..2034)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2189..2492
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2572..3160)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2652..2671)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3268..3316
                     /note="synthetic polyadenylation signal"
     misc_feature    3327..3418
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3367..3386
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        3455..3666
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3679..3947
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      3983..3992
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4744..4865
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    4991..5020
                     /label=NFAT binding site
                     /bound_moiety="NFAT transcription factor"
                     /note="NFAT binding site from the human interleukin 2 
                     (IL-2) gene (Mattila et al., 1990)"
     protein_bind    5021..5050
                     /label=NFAT binding site
                     /bound_moiety="NFAT transcription factor"
                     /note="NFAT binding site from the human interleukin 2 
                     (IL-2) gene (Mattila et al., 1990)"
     protein_bind    5051..5080
                     /label=NFAT binding site
                     /bound_moiety="NFAT transcription factor"
                     /note="NFAT binding site from the human interleukin 2 
                     (IL-2) gene (Mattila et al., 1990)"
     promoter        5093..5124
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      5180..5189
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5186..6838
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(5252..5270)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    6879..7000
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 aattctaaaa tacagcatag caaaacttta acctccaaat caagcctcta cttgaatcct
       61 tttctgaggg atgaataagg cataggcatc aggggctgtt gccaatgtgc attagctgtt
      121 tgcagcctca ccttctttca tggagtttaa gatatagtgt attttcccaa ggtttgaact
      181 agctcttcat ttctttatgt tttaaatgca ctgacctccc acattccctt tttagtaaaa
      241 tattcagaaa taatttaaat acatcattgc aatgaaaata aatgtttttt attaggcaga
      301 atccagatgc tcaaggccct tcataatatc ccccagttta gtagttggac ttagggaaca
      361 aaggaacctt taatagaaat tggacagcaa gaaagcgagc ttctagcttt agtcctgttc
      421 ctcagctaca aaatggacac aatttccagc agggtctctg agggcaaatt cccttcccca
      481 aggttgttca ccaatttctg tcatggctgg gccagaggca tccctgaaat ttgtgctgac
      541 tacttctgac cattctgcat aaagctcatc taggcctctg acccagaccc aagcaagggt
      601 gttgtcaggg acaacttggt cctgaactgc tgagatgaag agggtgacat catctctgac
      661 aacaccagca aaatcatctt caacaaagtc tctggagaat cctaatctgt cagtccagaa
      721 ctctacagcc cctgcaacat cccttgctgt gaggactggg actgcagaag tgagtttggc
      781 catgatggct cctcctgtca ggagaggaaa gagaagaagg ttagtacaat tgctatagtg
      841 agttgtatta tactatgctt atgattaatt gtcaaactag tgggttcata gtgccacttt
      901 tcctgcactg ccccatctcc tgcccaccct ttcccaggca tagacagtca gtgacttacc
      961 cttgtacagc tcatccattc ccagagtaat tcctgctgct gtcacaaact ccaggaggac
     1021 catgtggtct cttttctcat tagggtcttt ggacagagca gattgagtgc tgagatagtg
     1081 attatctggg aggagaactg ggccatcacc aataggggtg ttctgctggt aatggtctgc
     1141 cagttggaca gatccatcct caatgttgtg tctaatcttg aaattagcct taattccatt
     1201 cctctgctta tctgccataa tgtaaacatt gtgagaatta tagttgtact ccagcttgtg
     1261 acccagaatg tttccatctt ccttaaaatc aatgcctttc agctcaattc tgttaaccag
     1321 tgtatcacct tcaaacttca cttctgccct tgtcttataa tttccatcat ccttaaagaa
     1381 gattgtcctc tcctgaacat aaccttctgg cattgcagat ttaaagaagt catgctgctt
     1441 catgtggtca gggtatctgc tgaaacattg aacaccataa gtcagggtgg tcaccagagt
     1501 tggccaaggc actggcagct ttcctgttgt acaaatgaac ttcagagtca gctttccata
     1561 agttgcatct ccttcacctt caccagacac agagaatttg tggccattca catcaccatc
     1621 cagctcaacc agaattggga caacaccagt aaagagttct tctcccttag aaaccatggt
     1681 ggcttggatc tgtaacggcg cagaacagaa aacgaaacaa agacgtagag ttgagcaagc
     1741 agggtcaggc aaagcgtgga gagccggctg agtctaggta ggctccaagg gagcgccgga
     1801 caaaggcccg gtctcgacct gagctttaaa cttacctaga cggcggacgc agttcaggag
     1861 gcaccacagg cgggaggcgg cagaacgcga ctcaaccggc gtggatggcg gcctcaggta
     1921 gggcggcggg cgcgtgaagg agagatgcga gcccctcgaa gctgatctga cggttcacta
     1981 aacgagctct gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg
     2041 gggcggagtt gttacgacat tttggaaagt cccgttgatt tactagtcaa aacaaactcc
     2101 cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc
     2161 cattgatgta ctgccaaaac cgcatcatca tggtaatagc gatgactaat acgtagatgt
     2221 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt
     2281 taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca
     2341 agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg
     2401 ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc
     2461 caggcgggcc atttaccgta agttatgtaa cgcctgcagg ttaattaaga acatgtgagc
     2521 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2581 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2641 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2701 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2761 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2821 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2881 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2941 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3001 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3061 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3121 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3181 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatggctag
     3241 ttaattaaca tttaaatcag cggccgcaat aaaatatctt tattttcatt acatctgtgt
     3301 gttggttttt tgtgtgaatc gtaactaaca tacgctctcc atcaaaacaa aacgaaacaa
     3361 aacaaactag caaaataggc tgtccccagt gcaagtgcag gtgccagaac atttctctat
     3421 cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc
     3481 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg
     3541 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
     3601 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
     3661 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga
     3721 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga
     3781 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg
     3841 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct
     3901 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg
     3961 gcgcctacct gagatcaccg gtgccaccat ggccttacca gtgaccgcct tgctcctgcc
     4021 gctggccttg ctgctccacg ccgccaggcc gagccagttc cgggtgtcgc cgctggatcg
     4081 gacctggaac ctgggcgaga cagtggagct gaagtgccag gtgctgctgt ccaacccgac
     4141 gtcgggctgc tcgtggctct tccagccgcg cggcgccgcc gccagtccca ccttcctcct
     4201 atacctctcc caaaacaagc ccaaggcggc cgaggggctg gacacccagc ggttctcggg
     4261 caagaggttg ggggacacct tcgtcctcac cctgagcgac ttccgccgag agaacgaggg
     4321 ctactatttc tgctcggccc tgagcaactc catcatgtac ttcagccact tcgtgccggt
     4381 cttcctgcca gcgaagccca ccacgacgcc agcgccgcga ccaccaacac cggcgcccac
     4441 catcgcgtcg cagcccctgt ccctgcgccc agaggcgtgc cggccagcgg cggggggcgc
     4501 agtgcacacg agggggctgg acttcgcctg tgatatctac atctgggcgc ccttggccgg
     4561 gacttgtggg gtccttctcc tgtcactggt tatcaccctt tactgcaacc acaggaaccg
     4621 aagacgtgtt tgcaaatgtc cccggcctgt ggtcaaatcg ggagacaagc ccagcctttc
     4681 ggcgagatac gtccttaagg ggcccggcgc gcccgtacgg tatactcgcg actgcagccg
     4741 cggtaagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     4801 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     4861 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     4921 ttttttaaag caagtaaaac ctctacaaat gtggtatgga attcggtacc gagctcttac
     4981 gcgtgctagc ggaggaaaaa ctgtttcata cagaaggcgt ggaggaaaaa ctgtttcata
     5041 cagaaggcgt ggaggaaaaa ctgtttcata cagaaggcgt agatctagac tctagagggt
     5101 atataatgga agctcgaatt ccagcttggc attccggtac tgttggtaaa aagcttggca
     5161 ttccggtact gttggtaaag ccaccatgga agacgccaaa aacataaaga aaggcccggc
     5221 gccattctat ccgctggaag atggaaccgc tggagagcaa ctgcataagg ctatgaagag
     5281 atacgccctg gttcctggaa caattgcttt tacagatgca catatcgagg tggacatcac
     5341 ttacgctgag tacttcgaaa tgtccgttcg gttggcagaa gctatgaaac gatatgggct
     5401 gaatacaaat cacagaatcg tcgtatgcag tgaaaactct cttcaattct ttatgccggt
     5461 gttgggcgcg ttatttatcg gagttgcagt tgcgcccgcg aacgacattt ataatgaacg
     5521 tgaattgctc aacagtatgg gcatttcgca gcctaccgtg gtgttcgttt ccaaaaaggg
     5581 gttgcaaaaa attttgaacg tgcaaaaaaa gctcccaatc atccaaaaaa ttattatcat
     5641 ggattctaaa acggattacc agggatttca gtcgatgtac acgttcgtca catctcatct
     5701 acctcccggt tttaatgaat acgattttgt gccagagtcc ttcgataggg acaagacaat
     5761 tgcactgatc atgaactcct ctggatctac tggtctgcct aaaggtgtcg ctctgcctca
     5821 tagaactgcc tgcgtgagat tctcgcatgc cagagatcct atttttggca atcaaatcat
     5881 tccggatact gcgattttaa gtgttgttcc attccatcac ggttttggaa tgtttactac
     5941 actcggatat ttgatatgtg gatttcgagt cgtcttaatg tatagatttg aagaagagct
     6001 gtttctgagg agccttcagg attacaagat tcaaagtgcg ctgctggtgc caaccctatt
     6061 ctccttcttc gccaaaagca ctctgattga caaatacgat ttatctaatt tacacgaaat
     6121 tgcttctggt ggcgctcccc tctctaagga agtcggggaa gcggttgcca agaggttcca
     6181 tctgccaggt atcaggcaag gatatgggct cactgagact acatcagcta ttctgattac
     6241 acccgagggg gatgataaac cgggcgcggt cggtaaagtt gttccatttt ttgaagcgaa
     6301 ggttgtggat ctggataccg ggaaaacgct gggcgttaat caaagaggcg aactgtgtgt
     6361 gagaggtcct atgattatgt ccggttatgt aaacaatccg gaagcgacca acgccttgat
     6421 tgacaaggat ggatggctac attctggaga catagcttac tgggacgaag acgaacactt
     6481 cttcatcgtt gaccgcctga agtctctgat taagtacaaa ggctatcagg tggctcccgc
     6541 tgaattggaa tccatcttgc tccaacaccc caacatcttc gacgcaggtg tcgcaggtct
     6601 tcccgacgat gacgccggtg aacttcccgc cgccgttgtt gttttggagc acggaaagac
     6661 gatgacggaa aaagagatcg tggattacgt cgccagtcaa gtaacaaccg cgaaaaagtt
     6721 gcgcggagga gttgtgtttg tggacgaagt accgaaaggt cttaccggaa aactcgacgc
     6781 aagaaaaatc agagagatcc tcataaaggc caagaagggc ggaaagatcg ccgtgtaatt
     6841 ctagagtcgg ggcggccggc cgcttcgagc agacatgata agatacattg atgagtttgg
     6901 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     6961 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     7021 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     7081 caaatgtggt aaaatcgata aggatccgtc gacg
//
