LOCUS       sequence-379653-        6211 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of CnB (calcineurin subunit beta)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6211)
  AUTHORS   Cereghetti GM, Stangherlin A, Martins de Brito O, Chang CR, 
            Blackstone C, Bernardi P, Scorrano L
  TITLE     Dephosphorylation by calcineurin regulates translocation of Drp1 to 
            mitochondria.
  JOURNAL   Proc Natl Acad Sci U S A. 2008 Oct 14;105(41):15803-8. doi: 
            10.1073/pnas.0808249105. Epub 2008 Oct 6.
  PUBMED    18838687
REFERENCE   2  (bases 1 to 6211)
  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"; doi: 
            "10.1073/pnas.0808249105"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2008-10-14- 14"; volume: "105"; issue: "41"; pages: "15803-8"
COMMENT     Sequence Label: pcDNA3.1/Zeo-CnB
FEATURES             Location/Qualifiers
     source          1..6211
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    49..183
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(86..105)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     140..159
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(214..230)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(214..230)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(227..249)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    238..254
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(262..268,269..286,287..292))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    307..328
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(445..462)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(616..1204)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(696..715)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1375..2166,2167..2235))
                     /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     1998..2017
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2236..2340)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2415..2434)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2606..2985
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2986..3189
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3140..3160
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3234..3253
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3234..3252
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3960..3976
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3960..3976
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4010..4026)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4011..4027)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(4041..4059)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4042..4059)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4079..4096)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4085..4309
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4355..4868
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4442..4461)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4652..4673
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4891..4911)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4896..5225
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5076..5211
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5292..6086
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5346..5365)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5956..5975
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
       61 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
      121 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
      181 atccgtcgac ctcgagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
      241 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
      301 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
      361 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
      421 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
      481 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
      541 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      601 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      661 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      721 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      781 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      841 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      901 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      961 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1021 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     1081 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1141 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1201 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     1261 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     1321 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     1381 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1441 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     1501 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     1561 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1621 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     1681 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     1741 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     1801 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     1861 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1921 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     1981 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     2041 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     2101 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     2161 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     2221 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     2281 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     2341 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     2401 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     2461 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     2521 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
     2581 gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca
     2641 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     2701 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     2761 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     2821 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     2881 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     2941 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     3001 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     3061 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     3121 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     3181 agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg
     3241 actcactata gggagaccca agcttgtcga cggtaccccg gggaattcgg cagatcccga
     3301 ggtccgacag cgcccggccc agatccccac gcctgccagg agcaagccga gagccagccg
     3361 gccggcgcac tccgactccg agcagtctct gtccttcgac ccgagccccg cgccctttcc
     3421 gggacccctg ccccgcggat ccatgggaaa tgaggcaagt tatcctttgg aaatgtgctc
     3481 acactttgat gcggatgaaa ttaaaaggct aggaaagaga tttaagaagc ttgatttgga
     3541 caattctggt tctttgagtg tggaagagtt catgtctctg cctgagttac aacagaatcc
     3601 tttagtacag cgagtaatag atatattcga cacagatggg aatggagaag tagactttaa
     3661 agaattcatt gagggcgtct ctcagttcag tgtcaaagga gataaggagc agaaattgag
     3721 gtttgctttc cgtatctatg acatggataa agatggctat atttccaatg gggaactctt
     3781 ccaggtattg aagatgatgg tggggaacaa tctgaaagat acacagttac agcaaattgt
     3841 agacaaaacc ataataaatg cagataagga tggagatgga agaatatcct ttgaagaatt
     3901 ctgtgctgtt gtaggtggcc tagatatcca caaaaagatg gtggtagatg tgtgagaatt
     3961 ctagaactag tggatccccc gggctgcagg aattcgatat caagcttatc gataccgtcg
     4021 acctcgaggg ggggccctat tctatagtgt cacctaaatg ctagagctcg ctgatcagcc
     4081 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     4141 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     4201 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     4261 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     4321 gaaagaacca gctggggctc gaggggggat ccccacgcgc cctgtagcgg cgcattaagc
     4381 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4441 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4501 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4561 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4621 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4681 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     4741 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     4801 cttacaattt aaatatttgc ttatacaatc ttcctgtttt tggggctttt ctgattatca
     4861 accggggtgg gtaccgagct cgaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4921 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4981 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     5041 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     5101 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     5161 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     5221 gcaaaaagct cccgggagct tggatatcca ttttcggatc tgatcaagag acaggatgag
     5281 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     5341 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     5401 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     5461 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     5521 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     5581 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     5641 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     5701 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     5761 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     5821 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     5881 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     5941 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     6001 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     6061 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     6121 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     6181 cttcggaatc gttttccggg acgccggctg g
//
