LOCUS       sequence-169375-        6378 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 6378)
  AUTHORS   Macian F, Garcia-Rodriguez C, Rao A
  TITLE     Gene expression elicited by NFAT in the presence or absence of 
            cooperative recruitment of Fos and Jun.
  JOURNAL   EMBO J. 2000 Sep 1. 19(17):4783-95.
  PUBMED    10970869
REFERENCE   2  (bases 1 to 6378)
  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: "EMBO J. 
            2000 Sep 1. 19(17):4783-95."
COMMENT     Sequence Label: NFAT/AP-1 3x luciferase
FEATURES             Location/Qualifiers
     source          1..6378
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..63
                     /label=Mini-TK promoter
                     /note="minimal herpes simplex virus (HSV) thymidine kinase 
                     promoter"
     CDS             119..1771
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(185..203)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     intron          1902..1967
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2097..2117
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2542..2676
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3447..3465)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3533..3637
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3638..3706,3707..4498)
                     /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     complement(3856..3875)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4669..5257
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5158..5177
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5411..5428
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5443..5583
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5509..5528)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     polyA_signal    5822..5956
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    6050..6184
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 ttcgcatatt aaggtgacgc gtgtggcctc gaacaccgag cgaccctgca gcgacccgct
       61 taacagcgtc aacagcgtgc cgcagatctg agcttggcat tccggtactg ttggtaaaat
      121 ggaagacgcc aaaaacataa agaaaggccc ggcgccattc tatcctctag aggatggaac
      181 cgctggagag caactgcata aggctatgaa gagatacgcc ctggttcctg gaacaattgc
      241 ttttacagat gcacatatcg aggtgaacat cacgtacgcg gaatacttcg aaatgtccgt
      301 tcggttggca gaagctatga aacgatatgg gctgaataca aatcacagaa tcgtcgtatg
      361 cagtgaaaac tctcttcaat tctttatgcc ggtgttgggc gcgttattta tcggagttgc
      421 agttgcgccc gcgaacgaca tttataatga acgtgaattg ctcaacagta tgaacatttc
      481 gcagcctacc gtagtgtttg tttccaaaaa ggggttgcaa aaaattttga acgtgcaaaa
      541 aaaattacca ataatccaga aaattattat catggattct aaaacggatt accagggatt
      601 tcagtcgatg tacacgttcg tcacatctca tctacctccc ggttttaatg aatacgattt
      661 tgtaccagag tcctttgatc gtgacaaaac aattgcactg ataatgaatt cctctggatc
      721 tactgggtta cctaagggtg tggcccttcc gcatagaact gcctgcgtca gattctcgca
      781 tgccagagat cctatttttg gcaatcaaat cattccggat actgcgattt taagtgttgt
      841 tccattccat cacggttttg gaatgtttac tacactcgga tatttgatat gtggatttcg
      901 agtcgtctta atgtatagat ttgaagaaga gctgttttta cgatcccttc aggattacaa
      961 aattcaaagt gcgttgctag taccaaccct attttcattc ttcgccaaaa gcactctgat
     1021 tgacaaatac gatttatcta atttacacga aattgcttct gggggcgcac ctctttcgaa
     1081 agaagtcggg gaagcggttg caaaacgctt ccatcttcca gggatacgac aaggatatgg
     1141 gctcactgag actacatcag ctattctgat tacacccgag ggggatgata aaccgggcgc
     1201 ggtcggtaaa gttgttccat tttttgaagc gaaggttgtg gatctggata ccgggaaaac
     1261 gctgggcgtt aatcagagag gcgaattatg tgtcagagga cctatgatta tgtccggtta
     1321 tgtaaacaat ccggaagcga ccaacgcctt gattgacaag gatggatggc tacattctgg
     1381 agacatagct tactgggacg aagacgaaca cttcttcata gttgaccgct tgaagtcttt
     1441 aattaaatac aaaggatatc aggtggcccc cgctgaattg gaatcgatat tgttacaaca
     1501 ccccaacatc ttcgacgcgg gcgtggcagg tcttcccgac gatgacgccg gtgaacttcc
     1561 cgccgccgtt gttgttttgg agcacggaaa gacgatgacg gaaaaagaga tcgtggatta
     1621 cgtcgccagt caagtaacaa ccgcgaaaaa gttgcgcgga ggagttgtgt ttgtggacga
     1681 agtaccgaaa ggtcttaccg gaaaactcga cgcaagaaaa atcagagaga tcctcataaa
     1741 ggccaagaag ggcggaaagt ccaaattgta aaatgtaact gtattcagcg atgacgaaat
     1801 tcttagctat tgtaatactc tagaggatct ttgtgaagga accttacttc tgtggtgtga
     1861 cataattgga caaactacct acagagattt aaagctctaa ggtaaatata aaatttttaa
     1921 gtgtataatg tgttaaacta ctgattctaa ttgtttgtgt attttagatt ccaacctatg
     1981 gaactgatga atgggagcag tggtggaatg cctttaatga ggaaaacctg ttttgctcag
     2041 aagaaatgcc atctagtgat gatgaggcta ctgctgactc tcaacattct actcctccaa
     2101 aaaagaagag aaaggtagaa gaccccaagg actttccttc agaattgcta agttttttga
     2161 gtcatgctgt gtttagtaat agaactcttg cttgctttgc tatttacacc acaaaggaaa
     2221 aagctgcact gctatacaag aaaattatgg aaaaatattc tgtaaccttt ataagtaggc
     2281 ataacagtta taatcataac atactgtttt ttcttactcc acacaggcat agagtgtctg
     2341 ctattaataa ctatgctcaa aaattgtgta cctttagctt tttaatttgt aaaggggtta
     2401 ataaggaata tttgatgtat agtgccttga ctagagatca taatcagcca taccacattt
     2461 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     2521 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     2581 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     2641 tccaaactca tcaatgtatc ttatcatgtc tggatcgatc cccaggaagc tcctctgtgt
     2701 cctcataaac cctaacctcc tctacttgag aggacattcc aatcataggc tgcccatcca
     2761 ccctctgtgt cctcctgtta attaggtcac ttaacaaaaa ggaaattggg taggggtttt
     2821 tcacagaccg ctttctaagg gtaattttaa aatatctggg aagtcccttc cactgctgtg
     2881 ttccagaagt gttggtaaac agcccacaaa tgtcaacagc agaaacatac aagctgtcag
     2941 ctttgcacaa gggcccaaca ccctgctcat caagaagcac tgtggttgct gtgttagtaa
     3001 tgtgcaaaac aggaggcaca ttttccccac ctgtgtaggt tccaaaatat ctagtgtttt
     3061 catttttact tggatcagga acccagcact ccactggata agcattatcc ttatccaaaa
     3121 cagccttgtg gtcagtgttc atctgctgac tgtcaactgt agcatttttt ggggttacag
     3181 tttgagcagg atatttggtc ctgtagtttg ctaacacacc ctgcagctcc aaaggttccc
     3241 caccaacagc aaaaaaatga aaatttgacc cttgaatggg ttttccagca ccattttcat
     3301 gagttttttg tgtccctgaa tgcaagttta acatagcagt taccccaata acctcagttt
     3361 taacagtaac agcttcccac atcaaaatat ttccacaggt taagtcctca tttaaattag
     3421 gcaaaggaat tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     3481 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     3541 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3601 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     3661 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     3721 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     3781 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     3841 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca
     3901 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     3961 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4021 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4081 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4141 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt
     4201 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4261 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4321 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4381 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     4441 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     4501 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4561 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4621 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4681 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4741 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4801 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4861 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4921 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4981 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5041 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5101 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5161 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5221 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5281 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     5341 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     5401 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     5461 ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc
     5521 tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct
     5581 gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     5641 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     5701 tcatcaccga aacgcgcgag gcaggatcag ccatatcaca tttgtagagg ttttacttgc
     5761 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     5821 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5881 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5941 ttatcatgtc tggatcagcc atatcacatt tgtagaggtt ttacttgctt taaaaaacct
     6001 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgtta acttgtttat
     6061 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     6121 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     6181 gatccaagct cagatccaag cttggatcca agaggaaaat ttgtttcata cagaaggcgt
     6241 taagaggaaa atttgtttca tacagaaggc gttaagagga aaatttgttt catacagaag
     6301 gcgttcaagc ttgtcgactc gagatccggc gaattcgaac acgcagatgc agtcggggcg
     6361 gcgcggtccg aggtccac
//
