LOCUS       sequence-151440-        5267 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Full length RCAN1 expression with CMV promoter.  Flag tagged..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5267)
  AUTHORS   Liu Q, Busby JC, Molkentin JD
  TITLE     Interaction between TAK1-TAB1-TAB2 and RCAN1-calcineurin defines a 
            signalling nodal control point.
  JOURNAL   Nat Cell Biol. 2009 Feb;11(2):154-61. doi: 10.1038/ncb1823. Epub 
            2009 Jan 11.
  PUBMED    19136967
REFERENCE   2  (bases 1 to 5267)
  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: "10.1038/ncb1823"; 
            journalName: "Nat Cell Biol"; date: "2009-02"; volume: "11"; issue: 
            "2"; pages: "154-61"
COMMENT     Sequence Label: RCAN1
FEATURES             Location/Qualifiers
     source          1..5267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        250..629
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        630..833
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     784..804
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     830..854
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1064..1082
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1064..1081
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1095..1114
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1157..1180
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      1803..1812
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1816..1950
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1853..1872)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1907..1926
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2019..2376
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2019..2210
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2042..2062)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2227..2362
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2396..2413)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2396..2412)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2405..2427)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2625..3080
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2712..2731)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2922..2943
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3097..3116)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3216..3238
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3276..3294)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3362..3466
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3467..3535,3536..4327)
                     /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     complement(3685..3704)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4498..5086
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4987..5006
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5240..5257
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg aattaattcg
      241 agctcgcccg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
      301 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      361 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      421 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      481 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      541 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      601 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      661 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      721 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      781 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
      841 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
      901 gggaccgatc cagcctccgc ggccgggaac ggtgcattgg aacgcggatt ccccgtgcca
      961 agagtgacgt aagtaccgcc tatagagtct ataggcccac ccccttggct tcgttagaac
     1021 gcggctacaa ttaatacata accttatgta tcatacacat acgatttagg tgacactata
     1081 gaataacatc cactttgcct ttctctccac aggtgtccac tcccaggtcc aactgcacct
     1141 cggttctatc gatatggact acaaagacga tgacgacaag cgaattcccc ggggatccat
     1201 ggaggaggtg gatctgcagg acctgccgag cgccaccatc gcctgccacc tggacccgcg
     1261 cgtgttcgtg gacggcctgt gccgggccaa atttgaatcc ctcttcagaa catatgacaa
     1321 ggacaccacc ttccagtatt ttaagagctt caaacgtgtc cggataaact tcagcaaccc
     1381 cttatctgca gccgatgcca ggctgcggct gcacaagacc gagttcctgg ggaaggaaat
     1441 gaagttgtat tttgctcaga ctttacacat aggaagttca cacctggctc cgcccaatcc
     1501 cgacaaacag ttcctcatct cccctccggc ctctcctccc gttggctgga aacaagtaga
     1561 agatgccacc cccgtcataa attacgatct tttatatgcc atctccaagc tggggccagg
     1621 agagaagtat gaactgcatg cagcgacaga caccactccc agtgtggtgg tccacgtgtg
     1681 tgagagtgac caagagaatg aggaggaaga ggaagagatg gagagaatga agagacccaa
     1741 gcccaaaatc atccagacac ggagaccgga gtacacaccg atccacctta gctgaaagct
     1801 tggccgccat ggcccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     1861 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     1921 ctcatcaatg tatcttatca tgtctggatc gggaattaat tcggcgcagc accatggcct
     1981 gaaataacct ctgaaagagg aacttggtta ggtaccttct gaggcggaaa gaaccagctg
     2041 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2101 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     2161 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     2221 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     2281 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     2341 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tgttaacagc ttggcactgg
     2401 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     2461 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     2521 cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc
     2581 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     2641 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     2701 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     2761 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     2821 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     2881 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     2941 tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     3001 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     3061 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     3121 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     3181 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     3241 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     3301 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     3361 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3421 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     3481 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     3541 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     3601 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     3661 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     3721 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     3781 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     3841 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     3901 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     3961 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     4021 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     4081 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     4141 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     4201 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     4261 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     4321 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     4381 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4441 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4501 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4561 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4621 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4681 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4741 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4801 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4861 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4921 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4981 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5041 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5101 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     5161 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     5221 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaag
//
