LOCUS       sequence-129647-        4917 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 4917)
  TITLE     pBNP-NFAT-luciferase
REFERENCE   2  (bases 1 to 4917)
  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: pBNP-NFAT-luciferase
FEATURES             Location/Qualifiers
     source          1..4917
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(63..80)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(234..822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(314..333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(993..1784,1785..1853))
                     /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     1616..1635
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1854..1958)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1985..2440
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2072..2091)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2282..2303
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    2571..2619
                     /note="synthetic polyadenylation signal"
     misc_feature    2633..2724
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2673..2692
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        2825..2856
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      2912..2921
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2918..4570
                     /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(2984..3002)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    4611..4732
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4622..4641)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4676..4695
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctcttccgct
       61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     1861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1981 cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2041 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2101 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2161 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2221 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2281 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2341 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2401 tttaacgcga attttaacaa aatattaacg cttacaattt gccattcgcc attcaggctg
     2461 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gcccaagcta
     2521 ccatgataag taagtaatat taaggtacgg gaggtacttg gagcggccgc aataaaatat
     2581 ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagta ctaacatacg
     2641 ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc cccagtgcaa
     2701 gtgcaggtgc cagaacattt ctctatcgat aggtaccgag ctcttacgcg tctatccttt
     2761 tgttttccat cctgacgcgt ctatcctttt gttttccatc ctgcgcgtgc taggatctag
     2821 actctagagg gtatataatg gaagctcgaa ttccagcttg gcattccggt actgttggta
     2881 aaaagcttgg cattccggta ctgttggtaa agccaccatg gaagacgcca aaaacataaa
     2941 gaaaggcccg gcgccattct atccgctgga agatggaacc gctggagagc aactgcataa
     3001 ggctatgaag agatacgccc tggttcctgg aacaattgct tttacagatg cacatatcga
     3061 ggtggacatc acttacgctg agtacttcga aatgtccgtt cggttggcag aagctatgaa
     3121 acgatatggg ctgaatacaa atcacagaat cgtcgtatgc agtgaaaact ctcttcaatt
     3181 ctttatgccg gtgttgggcg cgttatttat cggagttgca gttgcgcccg cgaacgacat
     3241 ttataatgaa cgtgaattgc tcaacagtat gggcatttcg cagcctaccg tggtgttcgt
     3301 ttccaaaaag gggttgcaaa aaattttgaa cgtgcaaaaa aagctcccaa tcatccaaaa
     3361 aattattatc atggattcta aaacggatta ccagggattt cagtcgatgt acacgttcgt
     3421 cacatctcat ctacctcccg gttttaatga atacgatttt gtgccagagt ccttcgatag
     3481 ggacaagaca attgcactga tcatgaactc ctctggatct actggtctgc ctaaaggtgt
     3541 cgctctgcct catagaactg cctgcgtgag attctcgcat gccagagatc ctatttttgg
     3601 caatcaaatc attccggata ctgcgatttt aagtgttgtt ccattccatc acggttttgg
     3661 aatgtttact acactcggat atttgatatg tggatttcga gtcgtcttaa tgtatagatt
     3721 tgaagaagag ctgtttctga ggagccttca ggattacaag attcaaagtg cgctgctggt
     3781 gccaacccta ttctccttct tcgccaaaag cactctgatt gacaaatacg atttatctaa
     3841 tttacacgaa attgcttctg gtggcgctcc cctctctaag gaagtcgggg aagcggttgc
     3901 caagaggttc catctgccag gtatcaggca aggatatggg ctcactgaga ctacatcagc
     3961 tattctgatt acacccgagg gggatgataa accgggcgcg gtcggtaaag ttgttccatt
     4021 ttttgaagcg aaggttgtgg atctggatac cgggaaaacg ctgggcgtta atcaaagagg
     4081 cgaactgtgt gtgagaggtc ctatgattat gtccggttat gtaaacaatc cggaagcgac
     4141 caacgccttg attgacaagg atggatggct acattctgga gacatagctt actgggacga
     4201 agacgaacac ttcttcatcg ttgaccgcct gaagtctctg attaagtaca aaggctatca
     4261 ggtggctccc gctgaattgg aatccatctt gctccaacac cccaacatct tcgacgcagg
     4321 tgtcgcaggt cttcccgacg atgacgccgg tgaacttccc gccgccgttg ttgttttgga
     4381 gcacggaaag acgatgacgg aaaaagagat cgtggattac gtcgccagtc aagtaacaac
     4441 cgcgaaaaag ttgcgcggag gagttgtgtt tgtggacgaa gtaccgaaag gtcttaccgg
     4501 aaaactcgac gcaagaaaaa tcagagagat cctcataaag gccaagaagg gcggaaagat
     4561 cgccgtgtaa ttctagagtc ggggcggccg gccgcttcga gcagacatga taagatacat
     4621 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     4681 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     4741 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     4801 gtaaaacctc tacaaatgtg gtaaaatcga taaggatccg tcgaccgatg cccttgagag
     4861 ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcac
//
