LOCUS       sequence-133319-        6180 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 6180)
  AUTHORS   Kao SC, Wu H, Xie J, Chang CP, Ranish JA, Graef IA, Crabtree GR
  TITLE     Calcineurin/NFAT signaling is required for neuregulin-regulated 
            Schwann cell differentiation.
  JOURNAL   Science. 2009 Jan 30. 323(5914):651-4.
  PUBMED    19179536
REFERENCE   2  (bases 1 to 6180)
  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: "Science. 
            2009 Jan 30. 323(5914):651-4."
COMMENT     Sequence Label: Krox20 promoter pGL3-TATA
FEATURES             Location/Qualifiers
     source          1..6180
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1438..1447
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1444..3096
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(1510..1528)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3137..3258
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3148..3167)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3202..3221
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3506..3523)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3677..4265)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3757..3776)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4436..5227,5228..5296))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5059..5078
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5297..5401)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5428..5883
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5515..5534)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5725..5746
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6014..6062
                     /note="synthetic polyadenylation signal"
     misc_feature    6076..6167
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6116..6135
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 cgccgggaag ggcgcccgct ctgagcagca gctggagggg agggggcggg gagagcgcgg
       61 gcagagggag ggggcgcctc cggccgagga tctttctccc cgaagctccc gcagcccttc
      121 tgctccattc tctaccctaa cccctaccct caagttacct aaaaatactt aaacaaacaa
      181 acagcccaga cctgttccgt tttcactcct gtataaatag cacagacttt ccaaaaaagc
      241 aggaccgcat ttactcttat caccagacac tactttccac cgtgtaattc agaaaaaggc
      301 agtcagcttc cgtgagtgcg ggtaattttt ttctctctct ctttttttct ctgtctctct
      361 ctctcttttt ttcttttgct tgcggttttg agctgccaag aaagtgaagg aggggtttga
      421 ctgtagtgtc tcggcagcgc tcggttttct ttccgaagtt taattttccg gaatggctcc
      481 caaacaaggg ccggggaggc ggagccgcca ctaccggatc ttctcctttt ttggaaagtc
      541 tcggagaacc ggaattcctc cccgccccaa gagacagagc taccagcgcg gccgccgtgg
      601 gtgaactcac ggcggccgcg ctagggtcgg tgctcgcgcc ttcttcccgc tgaactctgc
      661 agtccggagt ccccgctgca ggcaggggcc gagagcccca gacccgggtg gttgtccacc
      721 ggctgcaaat cgttcctggc gagctcagcg gagcccgcgc agccaagccc gtatgcaaat
      781 tggccatgtg acggcaaaag ctgccaggcc cagccctgtt cctcagtcca tatatgggca
      841 gcgacgtcac gggtattgaa gacctgccca taaatactcc gagcctaaca ctttccgtct
      901 gagagagcag cgattgatta atagctgggc gaggggacac actgactgtt ataataacac
      961 tacaccagcg actcctggct ccccaacagc cggatcacag gcaggagaga gtcagtgacg
     1021 gatagacttt ttttttcttt aagaagccaa caacttggtt gctagtttta tttctgttaa
     1081 tttttttctt ttttttggtg tgtgtggatg tgttgtggtg gtcttttcta agtgtggagg
     1141 gcaaaaggag ataccatcct aggctcagtt caacccctct ccaaaaaaaa cggcttctct
     1201 ggcgctccag gtagcgaggg agttgggtct ccaggttgtg cgaggagcaa atgatgaccg
     1261 ccaaggccgt agacaaaatc ccagtaactc tcagtggttt tatgcaccag ctgagatcca
     1321 ctagcccggg ctcgagatct gcgatctaag taagcttctc gagcgggtct ccgtgacgac
     1381 tataaaagcc caggggcaag cggtccggaa gcttggcatt ccggtactgt tggtaaagcc
     1441 accatggaag acgccaaaaa cataaagaaa ggcccggcgc cattctatcc gctggaagat
     1501 ggaaccgctg gagagcaact gcataaggct atgaagagat acgccctggt tcctggaaca
     1561 attgctttta cagatgcaca tatcgaggtg gacatcactt acgctgagta cttcgaaatg
     1621 tccgttcggt tggcagaagc tatgaaacga tatgggctga atacaaatca cagaatcgtc
     1681 gtatgcagtg aaaactctct tcaattcttt atgccggtgt tgggcgcgtt atttatcgga
     1741 gttgcagttg cgcccgcgaa cgacatttat aatgaacgtg aattgctcaa cagtatgggc
     1801 atttcgcagc ctaccgtggt gttcgtttcc aaaaaggggt tgcaaaaaat tttgaacgtg
     1861 caaaaaaagc tcccaatcat ccaaaaaatt attatcatgg attctaaaac ggattaccag
     1921 ggatttcagt cgatgtacac gttcgtcaca tctcatctac ctcccggttt taatgaatac
     1981 gattttgtgc cagagtcctt cgatagggac aagacaattg cactgatcat gaactcctct
     2041 ggatctactg gtctgcctaa aggtgtcgct ctgcctcata gaactgcctg cgtgagattc
     2101 tcgcatgcca gagatcctat ttttggcaat caaatcattc cggatactgc gattttaagt
     2161 gttgttccat tccatcacgg ttttggaatg tttactacac tcggatattt gatatgtgga
     2221 tttcgagtcg tcttaatgta tagatttgaa gaagagctgt ttctgaggag ccttcaggat
     2281 tacaagattc aaagtgcgct gctggtgcca accctattct ccttcttcgc caaaagcact
     2341 ctgattgaca aatacgattt atctaattta cacgaaattg cttctggtgg cgctcccctc
     2401 tctaaggaag tcggggaagc ggttgccaag aggttccatc tgccaggtat caggcaagga
     2461 tatgggctca ctgagactac atcagctatt ctgattacac ccgaggggga tgataaaccg
     2521 ggcgcggtcg gtaaagttgt tccatttttt gaagcgaagg ttgtggatct ggataccggg
     2581 aaaacgctgg gcgttaatca aagaggcgaa ctgtgtgtga gaggtcctat gattatgtcc
     2641 ggttatgtaa acaatccgga agcgaccaac gccttgattg acaaggatgg atggctacat
     2701 tctggagaca tagcttactg ggacgaagac gaacacttct tcatcgttga ccgcctgaag
     2761 tctctgatta agtacaaagg ctatcaggtg gctcccgctg aattggaatc catcttgctc
     2821 caacacccca acatcttcga cgcaggtgtc gcaggtcttc ccgacgatga cgccggtgaa
     2881 cttcccgccg ccgttgttgt tttggagcac ggaaagacga tgacggaaaa agagatcgtg
     2941 gattacgtcg ccagtcaagt aacaaccgcg aaaaagttgc gcggaggagt tgtgtttgtg
     3001 gacgaagtac cgaaaggtct taccggaaaa ctcgacgcaa gaaaaatcag agagatcctc
     3061 ataaaggcca agaagggcgg aaagatcgcc gtgtaattct agagtcgggg cggccggccg
     3121 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     3181 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     3241 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     3301 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag
     3361 gatccgtcga ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg
     3421 gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca
     3481 ggtgccggca gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     3541 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3601 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3661 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3721 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3781 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3841 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3901 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3961 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4021 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4081 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     4141 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4201 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4261 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4321 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     4381 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     4441 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4501 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     4561 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4621 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4681 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4741 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4801 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4861 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4921 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4981 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5041 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5101 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     5161 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5221 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5281 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     5341 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     5401 gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg
     5461 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     5521 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     5581 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     5641 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     5701 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     5761 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     5821 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     5881 tttgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     5941 cgctattacg ccagcccaag ctaccatgat aagtaagtaa tattaaggta cgggaggtac
     6001 ttggagcggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg
     6061 tgaatcgata gtactaacat acgctctcca tcaaaacaaa acgaaacaaa acaaactagc
     6121 aaaataggct gtccccagtg caagtgcagg tgccagaaca tttctctatc gataggtacc
//
