LOCUS       sequence-174914-        6558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  rapamycin-mediated heterodimerization between PCRD and DCRD from 
            C-terminal of L-type Ca2+ channels.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6558)
  AUTHORS   Liu N, Yang Y, Ge L, Liu M, Colecraft HM, Liu X
  TITLE     Cooperative and acute inhibitionby multiple C-terminal motifs of 
            L-type Ca2+ channels.
  JOURNAL   Elife. 2017 Jan 6;6. pii: e21989. doi: 10.7554/eLife.21989.
  PUBMED    28059704
REFERENCE   2  (bases 1 to 6558)
  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: "Elife."; 
            date: "2017-01-6"; volume: "6. pii: e21989. doi"; pages: " 
            10.7554/eLife.21989"
COMMENT     Sequence Label: FKBP-DCRD-YFP
FEATURES             Location/Qualifiers
     source          1..6558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    683..845
                     /label=QBI SP163
                     /note="strong translational enhancer derived from the 5' 
                     UTR of the mouse VEGF gene"
     CDS             858..875
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             879..911
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(915..923,924..938)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(945..947,948..1268)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     CDS             join(1584..1586,1587..2300)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1626..1647)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1887..1906)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2234..2255
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2332..2349)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2338..2562
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2608..3036
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2695..2714)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2905..2926
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3045..3065)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3050..3379
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3230..3365
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3292..3311
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3427..3474
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3493..3867
                     /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    3997..4118
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4034..4053)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4088..4107
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4167..4183)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4167..4183)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4180..4202)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4191..4207
                     /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(4215..4221,4222..4239,4240..4245))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4260..4281
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4398..4415)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4569..5157)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4649..4668)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5328..6119,6120..6188))
                     /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     5951..5970
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6189..6293)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6368..6387)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttagcgcaga ggcttggggc agccgagcgg cagccaggcc
      721 ccggcccggg cctcggttcc agaagggaga ggagcccgcc aaggcgcgca agagagcggg
      781 ctgcctcgca gtccgagccg gagagggagc gcgagccgcg ccggccccgg acggcctccg
      841 aaaccatggg gggttctcat catcatcatc atcatggtat ggctagcatg actggtggac
      901 agcaaatggg tcgggatctg tacgacgatg acgataaggt accaatggga gtgcaggtgg
      961 aaaccatctc cccaggagac gggcgcacct tccccaagcg cggccagacc tgcgtggtgc
     1021 actacaccgg gatgcttgaa gatggaaaga aatttgattc ctcccgggac agaaacaagc
     1081 cctttaagtt tatgctaggc aagcaggagg tgatccgagg ctgggaagaa ggggttgccc
     1141 agatgagtgt gggtcagaga gccaaactga ctatatctcc agattatgcc tatggtgcca
     1201 ctgggcaccc aggcatcatc ccaccacatg ccactctcgt cttcgatgtg gagcttctaa
     1261 aactggaagg atccatgctg cacgtgcctg gaacccactc ggaccccagc catgggaaga
     1321 ggggcagtgc cgacagcttg gtggaggctg tgcttatctc agagggtctg ggcctctttg
     1381 ctcgagaccc acgtttcgtg gccctggcca agcaggagat tgcagatgcg tgtcgcctga
     1441 cgctggatga gatggacaat gctgccagtg acctgctggc acagggaacc agctctctct
     1501 atagcgacga ggagtccatc ctctcccgct tcgatgagga ggacttggga gacgagatgg
     1561 cctgcgtcca cgccctcgaa ttcgtgagca agggcgagga gctgttcacc ggggtggtgc
     1621 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     1681 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     1741 tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta cggcctgcag tgcttcgccc
     1801 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     1861 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     1921 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     1981 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     2041 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     2101 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     2161 tgctgctgcc cgacaaccac tacctgagct accagtccgc cctgagcaaa gaccccaacg
     2221 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     2281 tggacgagct gtacaagtag tctagagggc ccgtttaaac ccgctgatca gcctcgactg
     2341 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     2401 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     2461 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     2521 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     2581 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     2641 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     2701 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2761 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     2821 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2881 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     2941 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     3001 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     3061 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3121 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3181 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3241 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3301 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     3361 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     3421 gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg
     3481 aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc
     3541 gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac
     3601 gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag
     3661 gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc
     3721 gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag
     3781 atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg
     3841 cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc
     3901 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     3961 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     4021 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4081 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     4141 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4201 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4261 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4321 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4381 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4441 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4501 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4561 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4621 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4681 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4741 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4801 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4861 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4921 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4981 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5041 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5101 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5161 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5221 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5281 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5341 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5401 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5461 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5521 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5581 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5641 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5701 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5761 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5821 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5881 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5941 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6001 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6061 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6121 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6181 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6241 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6301 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
     6361 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     6421 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     6481 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
     6541 gggccagata tacgcgtt
//
