LOCUS       sequence-142525-        7496 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 7496)
  AUTHORS   Ichida M, Finkel T
  TITLE     Ras regulates NFAT3 activity in cardiac myocytes.
  JOURNAL   J Biol Chem. 2001 Feb 2. 276(5):3524-30.
  PUBMED    11044444
REFERENCE   2  (bases 1 to 7496)
  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: "J Biol 
            Chem. 2001 Feb 2. 276(5):3524-30."
COMMENT     Sequence Label: pEGFP-C1 NFAT3
FEATURES             Location/Qualifiers
     source          1..7496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      614..623
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(620..622,623..625,626..1336)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(620..622,623..625,626..1336)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(665..686)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(926..945)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1273..1294
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1337..1402
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    4291..4412
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4328..4347)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4382..4401
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4419..4874)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4556..4577)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4768..4787
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4901..5005
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5007..5364
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5007..5198
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5030..5050)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5215..5350
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5399..6193
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5399..6193
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5453..5472)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6063..6082
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6381..6400)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6425..6472
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6801..7389
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7290..7309
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
      661 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
      721 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
      781 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
      841 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
      901 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
      961 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     1021 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     1081 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     1141 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     1201 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     1261 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     1321 ggacgagctg tacaagtccg gactcagatc tcgagctcaa gcttcgaatt ctgcagtcga
     1381 cggtaccgcg ggcccgggat ccatgggggc ggccagctgc gaggatgagg agctggaatt
     1441 taagctggtg ttcggggagg aaaaggaggc ccccccgctg ggcgcggggg gattggggga
     1501 agaactggac tcagaggatg ccccgccatg ctgccgtctg gccttgggag agccccctcc
     1561 ctatggcgct gcacctatcg gtattccccg acctccaccc cctcggcctg gcatgcattc
     1621 gccaccgccg cgaccagccc cctcacctgg cacctgggag agccagcccg ccaggtcggt
     1681 gaggctggga ggaccaggag ggggtgctgg gggtgctggg ggtggccgtg ttctcgagtg
     1741 tcccagcatc cgcatcacct ccatctctcc cacgccggag ccgccagcag cgctggagga
     1801 caaccctgat gcctgggggg acggctctcc tagagattac cccccaccag aaggctttgg
     1861 gggctacaga gaagcagggg cccagggtgg gggggccttc ttcagcccaa gccctggcag
     1921 cagcagcctg tcctcgtgga gcttcttctc cgatgcctct gacgaggcag ccctgtatgc
     1981 agcctgcgac gaggtggagt ctgagctaaa tgaggcggcc tcccgctttg gcctgggctc
     2041 cccgctgccc tcgccccggg cctcccctcg gccatggacc cccgaagatc cctggagcct
     2101 gtatggtcca agccccggag gccgagggcc agaggatagc tggctactcc tcagtgctcc
     2161 tgggcccacc ccagcctccc cgcggcctgc ctctccatgt ggcaagcggc gctattccag
     2221 ctcgggaacc ccatcttcag cctccccagc tctgtcccgc cgtggcagcc tgggggaaga
     2281 ggggtctgag ccacctccac cacccccatt gcctctggcc cgggacccgg gctcccctgg
     2341 tccctttgac tatgtggggg ccccaccagc tgagagcatc cctcagaaga cacggcggac
     2401 ttccagcgag caggcagtgg ctctgcctcg gtctgaggag cctgcctcat gcaatgggaa
     2461 gctgcccttg ggagcagagg agtctgtggc tcctccagga ggttcccgga aggaggtggc
     2521 tggcatggac tacctggcag tgccctcccc actcgcttgg tccaaggccc ggattggggg
     2581 acacagccct atcttcagga cctctgccct acccccactg gactggcctc tgcccagcca
     2641 atatgagcag ctggagctga ggatcgaggt acagcctaga gcccaccacc gggcccacta
     2701 tgagacagaa ggcagccgtg gagctgtcaa agctgcccct ggcggtcacc ccgtagtcaa
     2761 gctcctaggc tacagtgaga agccactgac cctacagatg ttcatcggca ctgcagatga
     2821 aaggaacctg cggcctcatg ccttctatca ggtgcaccgt atcacaggca agatggtggc
     2881 cacggccagc tatgaagccg tagtcagtgg caccaaggtg ttggagatga ctctgctgcc
     2941 tgagaacaac atggcggcca acattgactg cgcgggaatc ctgaagcttc ggaattcaga
     3001 cattgagctt cggaagggtg agacggacat cgggcgcaaa aacacacgtg tacggctggt
     3061 gttccgggta cacgtgcccc agggcggcgg gaaggtcgtc tcagtacagg cagcatcggt
     3121 gcccatcgag tgctcccagc gctcagccca ggagctgccc caggtggagg cctacagccc
     3181 cagtgcctgc tctgtgagag gaggcgagga actggtactg accggctcca acttcctgcc
     3241 agactccaag gtggtgttca ttgagagggg tcctgatggg aagctgcaat gggaggagga
     3301 ggccacagtg aaccgactgc agagcaacga ggtgacgctg accctgactg tccccgagta
     3361 cagcaacaag agggtttccc ggccagtcca ggtctacttt tatgtctcca atgggcggag
     3421 gaaacgcagt cctacccaga gtttcaggtt tctgcctgtg atctgcaaag aggagcccct
     3481 accggactca tctctgcggg gtttcccttc agcatcggca accccctttg gcactgacat
     3541 ggacttctca ccacccaggc ccccctaccc ctcctatccc catgaagacc ctgcttgcga
     3601 aactccttac ctatcagaag gcttcggcta tggcatgccc cctctgtacc cccagacggg
     3661 gcccccacca tcctacagac cgggcctgcg gatgttccct gagactaggg gtaccacagg
     3721 ttgtgcccaa ccacctgcag tttccttcct tccccgcccc ttccctagtg acccgtatgg
     3781 agggcggggc tcctctttcc ccctggggct gccattctct ccgccagccc cctttcggcc
     3841 gcctcctctt cctgcatccc caccgcttga aggccccttc ccttcccaga gtgatgtgca
     3901 tcccctacct gctgagggat acaataaggt agggccaggc tatggccctg gggagggggc
     3961 tccggagcag gagaaatcca ggggtggcta cagcagcggc tttcgagaca gtgtccctat
     4021 ccagggtatc acgctggagg aagtgagtga gatcattggc cgagacctga gtggcttccc
     4081 tgcacctcct ggagaagagc ctcctgcctg aaccacgtga actgtcatca cctggcaacc
     4141 ccctcgtgcc gaattcgatg gggatccacc ggatctagat aactgatcat aatcagccat
     4201 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     4261 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     4321 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4381 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt
     4441 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     4501 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     4561 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     4621 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     4681 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     4741 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     4801 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     4861 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4921 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4981 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     5041 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5101 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5161 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     5221 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     5281 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     5341 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     5401 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     5461 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     5521 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     5581 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     5641 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     5701 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     5761 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     5821 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     5881 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     5941 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     6001 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     6061 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     6121 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     6181 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     6241 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     6301 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     6361 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     6421 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     6481 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     6541 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     6601 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     6661 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6721 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6781 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6841 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6901 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6961 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7021 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7081 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7141 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7201 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7261 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7321 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7381 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7441 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgc
//
