LOCUS       sequence-383850-        4852 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A FRET-based calcium probe using Sumire.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4852)
  AUTHORS   Sugiura K, Nagai T
  TITLE     Extension of the short wavelength side of fluorescent proteins using
            hydrated chromophores, and its application.
  JOURNAL   Commun Biol. 2022 Nov 3;5(1):1172. doi: 10.1038/s42003-022-04153-7.
  PUBMED    36329112
REFERENCE   2  (bases 1 to 4852)
  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: "Commun 
            Biol."; date: "2022-11-3"; volume: "5(1):1172. doi"; pages: " 
            10.1038/s42003-022-04153-7"
COMMENT     Sequence Label: vgCam-pRSETb
FEATURES             Location/Qualifiers
     source          1..4852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             70..92
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             112..129
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             133..165
                     /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(169..177,178..192)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1342..1419
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             2173..2196
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2253..2271)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2267..2314
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2411..2866
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2498..2517)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2708..2729
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2892..2996
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2997..3065,3066..3857)
                     /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(3215..3234)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4028..4616
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4517..4536
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4770..4787
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcggggttc tcatcatcat
      121 catcatcatg gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac
      181 gatgacgata aggatccaac catggtgagc aagggcgagg agctgttcac cggggtggtg
      241 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gcgcggcgag
      301 ggcgagggcg atgccaccaa cggcaagctg accctgaagt tcatctgcac caccggcaag
      361 ctgcccgtgc cctggcccac cctcgtgacc accctgggtt acggcgtggc gtgcttcagc
      421 cgctaccccg accacatgaa gcgccacgac ttcttcaagt ccgccatgcc cgaaggctac
      481 gtccaggagc gcaccatcag cttcaaggac gacggcacct acaagacccg cgccgaggtg
      541 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
      601 gacggcaaca tcctggggca caagctggag tacaacggta ttagcggcaa cgtgtatatc
      661 accgccgaca agcagaagaa cggcatcaag gccaactata agatccgcca caacgtggag
      721 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
      781 gtgctgctgc ccgacaacca ctacctgagc gtgcaggtgg tgctgagcaa agaccccaac
      841 gagaagcgcg atcacatggt cctgctggag ttccgtaccg ccgcccgcat gcatgaccaa
      901 ctgacagaag agcagattgc agagttcaaa gaagccttct cattattcga caaggatggg
      961 gacggcacca tcaccacaaa ggaacttggc accgttatga ggtcgcttgg acaaaaccca
     1021 acggaagcag aattgcagga tatgatcaat gaagtcgatg ctgatggcaa tggaacgatt
     1081 tactttcctg aatttcttac tatgatggct agaaaaatga aggacacaga cagcgaagag
     1141 gaaatccgag aagcattccg tgtttttgac aaggatggga acggctacat cagcgctgct
     1201 cagttacgtc acgtcatgac aaacctcggg gagaagttaa cagatgaaga agttgatgaa
     1261 atgataaggg aagcagatat cgatggtgat ggccaagtaa actatgaaga gtttgtacaa
     1321 atgatgacag caaagggggg gaagaggcgc tggaagaaaa acttcattgc cgtcagcgct
     1381 gccaaccggt tcaagaagat ctccagctcc ggggcactgg agctcatgga cggcagcgtg
     1441 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1501 gacaaccact acctgagcat ccagtccaag ctgagcaaag accccaacga gaagcgcgat
     1561 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     1621 tacaagggtg gcagcggtgg catggtgagc aagggcgagg agctgttcac cggggtggtg
     1681 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1741 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1801 ctgcccgtgc cctggcccac cctcgtgacc accttctcct acggcgtgat ggtgttcgcc
     1861 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1921 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1981 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2041 gacggcaaca tcctggggca caagctggag tacaacttca acagccacaa cgtctatatc
     2101 atggccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
     2161 gaattcggag gctggagcca cccgcagttc gaaaaataag cttgatccgg ctgctaacaa
     2221 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     2281 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatc
     2341 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     2401 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2461 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2521 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2581 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2641 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2701 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2761 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2821 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     2881 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2941 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     3001 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3061 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     3121 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3181 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3241 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3301 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3361 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3421 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3481 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3541 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     3601 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3661 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     3721 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3781 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3841 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     3901 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3961 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4021 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4081 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4141 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4201 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4261 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4321 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4381 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4441 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4501 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4561 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4621 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4681 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4741 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     4801 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc ag
//
