LOCUS       sequence-379808-        6109 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of phosphatase dead mutant of calcineurin A (CnA-H151A). 
            .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6109)
  AUTHORS   Scorrano L
  TITLE     Luca Scorrano plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 6109)
  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: 
            "unpublished"
COMMENT     Sequence Label: pcDNA3.1-CnA-H151A
FEATURES             Location/Qualifiers
     source          1..6109
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(40..628)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(120..139)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(799..1590,1591..1659))
                     /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     1422..1441
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1660..1764)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1839..1858)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2030..2409
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2410..2613
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2564..2584
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2658..2677
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2658..2676
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3912..3929)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3918..4142
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4188..4616
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4275..4294)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4485..4506
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4625..4645)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4630..4959
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4810..4945
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4872..4891
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5007..5054
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5073..5447
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    5577..5698
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5614..5633)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5668..5687
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5747..5763)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5747..5763)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5760..5782)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5771..5787
                     /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(5795..5801,5802..5819,5820..5825))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5840..5861
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5978..5995)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
       61 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
      121 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
      181 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
      241 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
      301 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
      361 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
      421 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
      481 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
      541 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
      601 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
      661 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
      721 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
      781 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
      841 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
      901 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
      961 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     1021 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     1081 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     1141 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     1201 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     1261 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     1321 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     1381 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     1441 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     1501 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     1561 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     1621 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     1681 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     1741 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     1801 atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca
     1861 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     1921 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
     1981 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
     2041 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     2101 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     2161 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     2221 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     2281 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     2341 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     2401 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     2461 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     2521 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     2581 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
     2641 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcatgtc
     2701 cgagcccaag gcgattgatc ccaagttgtc gactacggac agggtggtga aagccgttcc
     2761 atttccgcca agtcaccggc tgacagcaaa ggaagtgttt gataacgatg ggaagcctcg
     2821 tgtggatatc ttaaaagcac atctcatgaa ggaaggcagg ctggaagaaa gtgtcgcgtt
     2881 gagaataata acagagggtg cttcgattct ccgacaggaa aaaaacttgc tggatattga
     2941 tgccccagtc acagtttgcg gggacatcca tggacaattc tttgacttga tgaagctctt
     3001 tgaagtggga ggatctcctg ccaacactcg ctacctcttc ttaggggact atgttgacag
     3061 agggtacttc agtatcgaat gtgtgctgta tttgtgggcc ttgaaaattc tttaccccaa
     3121 aacactgttt ttacttcgtg gaaaccagga atgtaggcac ctaacagagt atttcacgtt
     3181 taaacaagaa tgtaaaataa agtattcaga acgcgtttat gacgcctgta tggatgcctt
     3241 cgactgcctt cccctggctg cgctgatgaa ccaacaattc ctgtgtgtac acggtggttt
     3301 gtctccagag attaacactc tagatgacat cagaaaatta gaccgattca aagaaccacc
     3361 tgcttatggg cctatgtgtg acatcttgtg gtcagacccc ctggaggact ttggaaatga
     3421 gaagactcag gaacatttca ctcacaacac agtcaggggt tgttcgtact tctacagtta
     3481 cccggctgta tgtgacttcc tgcagcacaa taatttgttg tccatactcc gagcccacga
     3541 agcccaggac gcagggtacc gcatgtacag gaaaagccaa acaactggct tcccgtctct
     3601 aattacgatc ttctcggcac caaattactt agatgtgtac aataataaag ctgcagtgtt
     3661 gaagtacgag aacaacgtga tgaacatcag gcagttcaac tgctcccccc atccgtactg
     3721 gctcccaaat ttcatggatg ttttcacctg gtcgctgcca tttgttgggg agaaagtgac
     3781 tgagatgctg gtaaacgtcc tgaacatctg ctcagatgat gaactggggt cagaagaaga
     3841 tggatttgac ggagccacgg ctgcagcccg gtgactcgag tctagagggc ccgtttaaac
     3901 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3961 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     4021 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     4081 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     4141 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     4201 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4261 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4321 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4381 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4441 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4501 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4561 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     4621 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4681 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4741 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4801 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4861 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4921 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4981 tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca
     5041 tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg
     5101 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
     5161 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
     5221 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
     5281 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
     5341 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
     5401 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag
     5461 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5521 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     5581 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5641 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     5701 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     5761 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5821 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5881 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5941 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6001 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6061 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagca
//
