LOCUS       sequence-323906-        6161 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 6161)
  TITLE     NFAT plasmids
REFERENCE   2  (bases 1 to 6161)
  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: pREP-NFAT2
FEATURES             Location/Qualifiers
     source          1..6161
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..330,331..345,346..614)
                     /label=SR-alpha promoter
                     /note="hybrid promoter consisting of the SV40 early 
                     promoter plus part of the long terminal repeat of human 
                     T-lymphotrophic virus 1"
     rep_origin      181..316
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     LTR             346..614
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     intron          658..754
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             complement(920..931)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3566..3700
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3590..3609)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3644..3663
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3843..3865)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3951..4091
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4006..4025
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4106..4123)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4277..4865)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4357..4376)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5036..5827,5828..5896))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5659..5678
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5897..6001)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6069..6087
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(join(6157..6161,1..16))
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
       61 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
      121 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
      181 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
      241 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
      301 ggcttttttg gaggcctagg cttttgcaaa aagctcctcg atcgaggggc tcgcatctct
      361 ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg
      421 ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc
      481 aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc
      541 tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg
      601 cgccgttaca gatccaagct cctcgatcga ggaactgaaa aaccagaaag ttaactggta
      661 agtttagtct ttttgtcttt tatttcaggt cccggatccg gtggtggtgc aaatcaaaga
      721 actgctcctc agtggatgtt gcctttactt ctaggcctgt acggaagtgt tacttctgct
      781 ctaaaagctg ctctagccct cgagggctag agcggccgcg gaggccgaat tccgcagggc
      841 gcgggcaccg gggcgcgggc agggctcgga gccaccgcgc aggtcctagg gccgcggccg
      901 ggccccgcca cgcgcgcaca cgcccctcga tgactttcct ccggggcgcg cggcgctgag
      961 cccggggcga gggctgtctt cccggagacc cgaccccggc agcgcggggc ggccacttct
     1021 cctgtgcctc cgcccgctgc tccactcccc gccgccgccg cgcggatgcc aagcaccagc
     1081 tttccagtcc cttccaagtt tccacttggc cctgcggctg cggtcttcgg gagaggagaa
     1141 actttggggc ccgcgccgcg cgccggcggc accatgaagt cagcggagga agaacactat
     1201 ggctatgcat cctccaacgt cagccccgcc ctgccgctcc ccacggcgca ctccaccctg
     1261 ccggccccgt gccacaacct tcagacctcc acaccgggca tcatcccgcc ggcggatcac
     1321 ccctcggggt acggagcagc tttggacggt gggcccgcgg gctacttcct ctcctccggc
     1381 cacaccaggc ctgatggggc ccctgccctg gagagtcctc gcatcgagat aacctcgtgc
     1441 ttgggcctgt accacaacaa taaccagttt ttccacgatg tggaggtgga agacgtcctc
     1501 cctagctcca aacggtcccc ctccacggcc acgctgagtc tgcccagcct ggaggcctac
     1561 agagacccct cgtgcctgag cccggccagc agcctgtcct cccggagctg caactcagag
     1621 gcctcctcct acgagtccaa ctactcgtac ccgtacgcgt ccccccagac gtcgccatgg
     1681 cagtctccct gcgtgtctcc caagaccacg gaccccgagg agggctttcc ccgcgggctg
     1741 ggggcctgca cactgctggg ttccccgcag cactccccct ccacctcgcc ccgcgccagc
     1801 gtcactgagg agagctggct gggtgcccgc tcctccagac ccgcgtcccc ttgcaacaag
     1861 aggaagtaca gcctcaacgg ccggcagccg ccctactcac cccaccactc gcccacgccg
     1921 tccccgcacg gctccccgcg ggtcagcgtg accgacgact cgtggttggg caacaccacc
     1981 cagtacacca gctcggccat cgtggccgcc atcaacgcgc tgaccaccga cagcagcctg
     2041 gacctgggag atggcgtccc tgtcaagtcc cgcaagacca ccctggagca gccgccctca
     2101 gtggcgctca aggtggagcc cgtcggggag gacctgggca gccccccgcc cccggccgac
     2161 ttcgcgcccg aagactactc ctctttccag cacatcagga agggcggctt ctgcgaccag
     2221 tacctggcgg tgccgcagca cccctaccag tgggcgaagc ccaagcccct gtcccctacg
     2281 tcctacatga gcccgaccct gcccgccctg gactggcagc tgccgtccca ctcaggcccg
     2341 tatgagcttc ggattgaggt gcagcccaag tcccaccacc gagcccacta cgagacggag
     2401 ggcagccggg gggccgtgaa ggcgtcggcc ggaggacacc ccatcgtgca gctgcatggc
     2461 tacttggaga atgagccgct gatgctgcag cttttcattg ggacggcgga cgaccgcctg
     2521 ctgcgcccgc acgccttcta ccaggtgcac cgcatcacag ggaagaccgt gtccaccacc
     2581 agccacgagg ctatcctctc caacaccaaa gtcctggaga tcccactcct gccggagaac
     2641 agcatgcgag ccgtcattga ctgtgccgga atcctgaaac tcagaaactc cgacattgaa
     2701 cttcggaaag gagagacgga catcgggagg aagaacacac gggtacggct ggtgttccgc
     2761 gttcacgtcc cgcaacccag cggccgcacg ctgtccctgc aggtggcctc caaccccatc
     2821 gaatgctccc agcgctcagc tcaggagctg cctctggtgg agaagcagag cacggacagc
     2881 tatccggtcg tgggcgggaa gaagatggtc ctgtctggcc acaacttcct gcaggactcc
     2941 aaggtcattt tcgtggagaa agccccagat ggccaccatg tctgggagat ggaagcgaaa
     3001 actgaccggg acctgtgcaa gccgaattct ctggtggttg agatcccgcc atttcggaat
     3061 cagaggataa ccagccccgt tcacgtcagt ttctacgtct gcaacgggaa gagaaagcga
     3121 agccagtacc agcgtttcac ctaccttccc gccaacggta acgccatctt tctaaccgta
     3181 agccgtgaac atgagcgcgt ggggtgcttt ttctaaagac gcagaaacga cgtcgccgta
     3241 aagcagcgtg gcgtgttgca catttaactg tgtgatgtcc cgttagtgag accgagccat
     3301 cgatgccctg aaaaggaaag gaaaagggaa gcttcggatg cattttcctt gatccctgtt
     3361 gggggtgggg ggcgggggtt gcatactcag atagtcacgg ttattttgct tcttgcgaat
     3421 gtataacagc caaggggaaa acatggctct tctgctccaa aaaactgagg gggtcctggt
     3481 gtgcatttgc accctaaagc tgcttacggt gaaaaggcaa ataggtatag ctattttgca
     3541 ggcaccttta ggaataaact ttgcggatcc agacatgata agatacattg atgagtttgg
     3601 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     3661 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     3721 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     3781 caaatgtggt atggctgatt atgatccggc tgcctcgcgc gtttcggtga tgacggtgaa
     3841 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     3901 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg
     3961 acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga
     4021 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     4081 accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4141 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4201 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4261 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4321 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4381 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4441 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4501 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4561 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4621 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4681 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     4741 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4801 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4861 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4921 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     4981 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5041 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5101 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5161 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5221 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     5281 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5341 ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5401 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5461 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     5521 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     5581 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5641 gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5701 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     5761 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5821 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5881 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     5941 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6001 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     6061 cctataaaaa taggcgtatc acgaggccct ttcgtcttca agaattggtc gaccaattct
     6121 catgtttgac agcttatcat cgataagctt ggctgtggaa t
//
