LOCUS       sequence-7061-ex        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      103..189
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     316..339
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    358..457
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             502..591
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     protein_bind    complement(3418..3517)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3629..3648)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3640..4449
                     /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(3707..3726)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4539..5127
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5028..5047
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggcttcg aaggagatag aaccaattct
      481 taagcttcca ccatgatctt ttacccatac gatgttcctg actatgcggg ctatccctat
      541 gacgtcccgg actatgcagg atcctatcca tatgacgttc cagattacgc tgctcagtgc
      601 ggccgcgacg tcccggagcc gcagcccgac cccgatggcg gggacggccc cggccacgag
      661 cccgggggca gtccccaaga cgagctggac ttttccatcc tcttcgatta tgactatctg
      721 aaccctatcg aagaagaacc gatcgcacat aaggccatca gctcaccctc cggactcgca
      781 tacccggatg atgtcctgga ctatggcctc aagccatgca acccccttgc cagtctctct
      841 ggcgagcccc ctggccggtt cggagagccg gatagtatag ggttccagaa ctttctgagc
      901 ccggtcaagc cagcaggggc ttcgggcccg agccctcgga tcgagatcac tccatcccac
      961 gaactgatgc aggcaggggg ggccctccgt gggagagacg ccggcctgtc ccccgagcag
     1021 ccggccctgg ccctggccgg cgtggccgcc agcccgaggt tcacactgcc cgtgcccggc
     1081 tacgagggct accgcgagcc gctttgcttg agccccgctg cagcaggcgc agcagccgca
     1141 ttcatttctg acaccttctc cccctacacc tcgccctgcg tctcacccaa taacgccggg
     1201 cccgacgacc tgtgtcccca gtttcaaaac atccctgctc attatgcccc ccggaccgct
     1261 ccaataatgg cacctcgaac cagcctcgcc gaggacagct gcctgggccg acactcgccc
     1321 gtgccccgtc cggcatcccg ctcctcctca cccggtgcca agcggaggca ttcgtgcgca
     1381 gaggctttgg ttgctcctct gcccgcagcc gcaccccagc gcgcccggag ccccgcgcca
     1441 cagcccgcgc ctcacgtggc accgcaggac gacagcatcc ccgctgggta cccccccacg
     1501 gccggctctg ctgttctcat ggatgccctc aacaccctgg ccaccgactc gccctgcggg
     1561 atcccctcca agatatggaa gaccagtcct gacccgacgc ctgtgtccac cgctccgtcc
     1621 aaggctggcc tggcccgcca catctaccct actgtggagt tcctggggcc atgtgagcag
     1681 gaggagagga ggaattccgc tccagagtcc atcctgctgg taccacctac ttggcccaag
     1741 cagttggtgc cggccattcc catctgcagc atccctgtga ctgcatccct cccaccactc
     1801 gagtggccac tctccaatca gtcgggctcc tatgagctac ggattgaggt ccaacccaag
     1861 ccccatcacc gggcccacta tgagacggag ggcagccgtg gcgctgtcaa agccccaaca
     1921 ggaggacacc ctgtggtgca gctccacggc tacatggaga acaagcctct ggggcttcag
     1981 atcttcattg ggacagcaga tgagaggatc cttaagccgc acgccttcta ccaagtacac
     2041 aggatcactg ggaaaacggt caccaccacg agctatgaga agatcgtagg caacaccaag
     2101 gtcctggaga tccccctgga gccaaagaac aacatgagag ccaccatcga ctgtgcaggc
     2161 atcctgaagc tccgaaacgc tgacatcgag ctgcggaagg gcgagacgga catcggcagg
     2221 aagaacacgc gtgtgcgcct ggtgttccgc gtgcacgtcc cagagcccag tgggcgcatc
     2281 gtctccctgc aggctgcgtc caaccccatc gagtgctctc agcgctctgc ccacgagctg
     2341 cccatggtgg agagacaaga catggacagc tgcctggtct acgggggcca gcagatgatc
     2401 ctcacgggcc agaacttcac agcggagtcc aaggttgtgt tcatggagaa gactacagat
     2461 gggcagcaga tttgggagat ggaagctacg gtggataaag acaagagcca gcctaacatg
     2521 ctttttgttg agatccccga gtatcggaac aagcacatcc gcgtgcccgt gaaagtcaac
     2581 ttctacgtca tcaacggaaa gaggaaacga agtcagccac agcactttac ctaccaccca
     2641 gtccctgcca tcaagacaga gcccagcgat gagtatgaac catctttgat ctgcagcccc
     2701 gcccatggag gcctggggag ccagccatat tacccacagc acccaatgct ggccgagtcc
     2761 ccctcctgcc ttgtggctac catggccccc tgccaacagt tccgctcggg gctctcatcc
     2821 cccgatgctc gctaccaaca gcagagcccc gcagctgccc tctaccagag aagcaagagc
     2881 ctgagtcccg gcctgctggg ctaccagcag ccgtccctcc tggcagcacc cttgggtctg
     2941 gctgatgccc accgctctgt gctggtgcat gctggttctc aggggcaggg gcagggctcc
     3001 accctccgac acacatcctc ggccagccag caggcctcac ccgtgatcca ctactcaccc
     3061 accaaccagc agcttcgcgg tgggggtcac caggagttcc agcatatcat gtactgtgaa
     3121 aacttcggcc ccagctctgc caggcctggc ccgcctccca tcaaccaagg tcagaggctg
     3181 agcccgggcg cctaccccac agtcatccaa caacagactg ccccgagcca aagagctgcc
     3241 aaaaacggac ccagtgacca gaaggaagct ctgcccacgg gagtgaccgt caaacaggaa
     3301 cagaacctgg accagaccta cctggatgac gttaatgaaa tcatcaggaa ggagttttca
     3361 ggacctccct cccgaaatca gacctagaaa caaacataat ctcgcgacgc aatctagacc
     3421 cagctttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg
     3481 aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgca gctctggccc
     3541 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
     3601 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     3661 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     3721 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
     3781 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     3841 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     3901 actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta
     3961 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     4021 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
     4081 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     4141 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     4201 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     4261 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     4321 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     4381 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     4441 tttttctaat cagaattggt taattggttg taacattatt cagattgggc cccgttccac
     4501 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4561 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4621 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     4681 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     4741 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     4801 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     4861 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4921 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     4981 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5041 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5101 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5161 gccttttgct ggccttttgc tcacatgtt
//
