LOCUS       sequence-229776-        5189 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5189)
  TITLE     NFAT plasmids
REFERENCE   2  (bases 1 to 5189)
  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: pENTR11 CA NFAT1
FEATURES             Location/Qualifiers
     source          1..5189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      105..191
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      283..310
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     318..341
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    360..459
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             504..593
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     protein_bind    complement(3420..3519)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3631..3650)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3642..4451
                     /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(3709..3728)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4541..5129
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5030..5049
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgctag catggatctc
       61 ggggacgtct aactactaag cgagagtagg gaactgccag gcatcaaata aaacgaaagg
      121 ctcagtcgga agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
      181 gtaggacaaa tccgccggga gcggatttga acgttgtgaa gcaacggccc ggagggtggc
      241 gggcaggacg cccgccataa actgccaggc atcaaactaa gcagaaggcc atcctgacgg
      301 atggcctttt tgcgtttcta caaactcttc ctgttagtta gttacttaag ctcgggcccc
      361 aaataatgat tttattttga ctgatagtga cctgttcgtt gcaacaaatt gataagcaat
      421 gcttttttat aatgccaact ttgtacaaaa aagcaggctt cgaaggagat agaaccaatt
      481 cttaagcttc caccatgatc ttttacccat acgatgttcc tgactatgcg ggctatccct
      541 atgacgtccc ggactatgca ggatcctatc catatgacgt tccagattac gctgctcagt
      601 gcggccgcga cgtcccggag ccgcagcccg accccgatgg cggggacggc cccggccacg
      661 agcccggggg cagtccccaa gacgagctgg acttttccat cctcttcgat tatgactatc
      721 tgaaccctat cgaagaagaa ccgatcgcac ataaggccat cagctcaccc tccggactcg
      781 catacccgga tgatgtcctg gactatggcc tcaagccatg caaccccctt gccagtctct
      841 ctggcgagcc ccctggccgg ttcggagagc cggatagtat agggttccag aactttctga
      901 gcccggtcaa gccagcaggg gcttcgggcc cgagccctcg gatcgagatc actccatccc
      961 acgaactgat gcaggcaggg ggggccctcc gtgggagaga cgccggcctg tcccccgagc
     1021 agccggccct ggccctggcc ggcgtggccg ccagcccgag gttcacactg cccgtgcccg
     1081 gctacgaggg ctaccgcgag ccgctttgct tgagccccgc tgcagcaggc gcagcagccg
     1141 cattcatttc tgacaccttc tccccctaca cctcgccctg cgtctcaccc aataacgccg
     1201 ggcccgacga cctgtgtccc cagtttcaaa acatccctgc tcattatgcc ccccggaccg
     1261 ctccaataat ggcacctcga accagcctcg ccgaggacag ctgcctgggc cgacactcgc
     1321 ccgtgccccg tccggcatcc cgctcctcct cacccggtgc caagcggagg cattcgtgcg
     1381 cagaggcttt ggttgctcct ctgcccgcag ccgcacccca gcgcgcccgg agccccgcgc
     1441 cacagcccgc gcctcacgtg gcaccgcagg acgacagcat ccccgctggg taccccccca
     1501 cggccggctc tgctgttctc atggatgccc tcaacaccct ggccaccgac tcgccctgcg
     1561 ggatcccctc caagatatgg aagaccagtc ctgacccgac gcctgtgtcc accgctccgt
     1621 ccaaggctgg cctggcccgc cacatctacc ctactgtgga gttcctgggg ccatgtgagc
     1681 aggaggagag gaggaattcc gctccagagt ccatcctgct ggtaccacct acttggccca
     1741 agcagttggt gccggccatt cccatctgca gcatccctgt gactgcatcc ctcccaccac
     1801 tcgagtggcc actctccaat cagtcgggct cctatgagct acggattgag gtccaaccca
     1861 agccccatca ccgggcccac tatgagacgg agggcagccg tggcgctgtc aaagccccaa
     1921 caggaggaca ccctgtggtg cagctccacg gctacatgga gaacaagcct ctggggcttc
     1981 agatcttcat tgggacagca gatgagagga tccttaagcc gcacgccttc taccaagtac
     2041 acaggatcac tgggaaaacg gtcaccacca cgagctatga gaagatcgta ggcaacacca
     2101 aggtcctgga gatccccctg gagccaaaga acaacatgag agccaccatc gactgtgcag
     2161 gcatcctgaa gctccgaaac gctgacatcg agctgcggaa gggcgagacg gacatcggca
     2221 ggaagaacac gcgtgtgcgc ctggtgttcc gcgtgcacgt cccagagccc agtgggcgca
     2281 tcgtctccct gcaggctgcg tccaacccca tcgagtgctc tcagcgctct gcccacgagc
     2341 tgcccatggt ggagagacaa gacatggaca gctgcctggt ctacgggggc cagcagatga
     2401 tcctcacggg ccagaacttc acagcggagt ccaaggttgt gttcatggag aagactacag
     2461 atgggcagca gatttgggag atggaagcta cggtggataa agacaagagc cagcctaaca
     2521 tgctttttgt tgagatcccc gagtatcgga acaagcacat ccgcgtgccc gtgaaagtca
     2581 acttctacgt catcaacgga aagaggaaac gaagtcagcc acagcacttt acctaccacc
     2641 cagtccctgc catcaagaca gagcccagcg atgagtatga accatctttg atctgcagcc
     2701 ccgcccatgg aggcctgggg agccagccat attacccaca gcacccaatg ctggccgagt
     2761 ccccctcctg ccttgtggct accatggccc cctgccaaca gttccgctcg gggctctcat
     2821 cccccgatgc tcgctaccaa cagcagagcc ccgcagctgc cctctaccag agaagcaaga
     2881 gcctgagtcc cggcctgctg ggctaccagc agccgtccct cctggcagca cccttgggtc
     2941 tggctgatgc ccaccgctct gtgctggtgc atgctggttc tcaggggcag gggcagggct
     3001 ccaccctccg acacacatcc tcggccagcc agcaggcctc acccgtgatc cactactcac
     3061 ccaccaacca gcagcttcgc ggtgggggtc accaggagtt ccagcatatc atgtactgtg
     3121 aaaacttcgg ccccagctct gccaggcctg gcccgcctcc catcaaccaa ggtcagaggc
     3181 tgagcccggg cgcctacccc acagtcatcc aacaacagac tgccccgagc caaagagctg
     3241 ccaaaaacgg acccagtgac cagaaggaag ctctgcccac gggagtgacc gtcaaacagg
     3301 aacagaacct ggaccagacc tacctggatg acgttaatga aatcatcagg aaggagtttt
     3361 caggacctcc ctcccgaaat cagacctaga aacaaacata atctcgcgac gcaatctaga
     3421 cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa
     3481 cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg cagctctggc
     3541 ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa
     3601 taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg
     3661 aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg
     3721 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg
     3781 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga
     3841 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc
     3901 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg
     3961 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc
     4021 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc
     4081 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg
     4141 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct
     4201 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg
     4261 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
     4321 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
     4381 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg
     4441 agtttttcta atcagaattg gttaattggt tgtaacatta ttcagattgg gccccgttcc
     4501 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4561 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4621 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4681 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4741 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4801 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4861 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4921 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4981 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5041 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5101 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5161 tggccttttg ctggcctttt gctcacatg
//
