LOCUS       Exported                4455 bp DNA     circular SYN 19-SEP-2025
DEFINITION  Red intensiometric genetically encoded Ca2+-indicators for optical 
            imaging 1.2.
ACCESSION   .
VERSION     .
KEYWORDS    CMV-R-GECO1.2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4455)
  AUTHORS   Wu J, Liu L, Matsuda T, Zhao Y, Rebane A, Drobizhev M, Chang YF, 
            Araki S, Arai Y, March K, Hughes TE, Sagou K, Miyata T, Nagai T, Li 
            WH, Campbell RE
  TITLE     Improved Orange and Red Ca Indicators and Photophysical 
            Considerations for Optogenetic Applications.
  JOURNAL   ACS Chem Neurosci. 2013 Mar 19.
  PUBMED    23452507
REFERENCE   2  (bases 1 to 4455)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 19, 2025 from SnapGene 6.0.2
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Chem 
            Neurosci. 2013 Mar 19."
FEATURES             Location/Qualifiers
     source          1..4455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        64..443
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        444..647
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     598..618
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     692..711
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        692..710
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             745..1998
                     /codon_start=1
                     /label=R-GECO1.2
                     /translation="MVDSSRRKWNKAGHAVRAIGRLSSPVVSERMYPEDGALKSEIKKG
                     LRLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQCERAEGRHST
                     GGMDELYKGGTGGSLVSKGEEDNRAIVKEFMRFKLHMEGSVNGHEFEIEGEGEGRPYEA
                     FQTAKLKVTKGGPLPFAWDILSPQLMYGSKAYIKHPADIPDYFKLSFPEGFRWERVMNF
                     EDGGIIHVSQDTSLQDGVFIYKVKLRGTNFPPDGPVMQKKTMGWEATRDQLTEEQIAEF
                     KEAFSLFDKDGDGTMTTKELGTVMRSLGQNPTEAELQDMINEVDADGDGTFDFPEFLTM
                     MARKMNDTDSEEEIREAFRVFDKDGNGYIGAAELRHVMTDLGEKITDEEVDEMIRVADI
                     DGDGQVNYEEFVQMMTAK"
     CDS             760..816
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     1512..1531
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        complement(2059..2077)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2060..2077)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2097..2114)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2103..2327
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2529..3117)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2609..2628)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3288..4148)
                     /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     3911..3930
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4149..4253)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4328..4347)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
       61 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
      121 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      181 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      241 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      301 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      361 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      421 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      481 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      541 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      601 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta
      661 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag
      721 cttggtaccg agctcggatc caccatggtc gactcatcac gtcgtaagtg gaataaggca
      781 ggtcacgcag tcagagctat aggtcggctg agctcacccg tggtttccga gcggatgtac
      841 cccgaggacg gcgccctgaa gagcgagatc aagaaggggc tgaggctgaa ggacggcggc
      901 cactacgccg ccgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
      961 gcctacatcg tcgacatcaa gttggacatc gtgtcccaca acgaggacta caccatcgtg
     1021 gaacagtgcg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     1081 ggaggtacag gcgggagtct ggtgagcaag ggcgaggagg ataacagggc catcgtcaag
     1141 gagttcatgc gcttcaagtt gcacatggag ggctccgtga acggccacga gttcgagatc
     1201 gagggcgagg gcgagggccg cccctacgag gcctttcaga ccgctaagct gaaggtgacc
     1261 aagggtggcc ccctgccctt cgcatgggac atcctgtccc ctcagttgat gtacggctcc
     1321 aaggcctaca ttaagcaccc agccgacatc cccgactact tcaagctgtc cttccccgag
     1381 ggcttcaggt gggagcgcgt gatgaacttc gaggacggcg gcattattca cgttagccag
     1441 gacacctccc tgcaggacgg cgtattcatc tacaaggtga agctgcgcgg caccaacttc
     1501 ccccccgacg gccccgtaat gcagaagaag accatgggct gggaggctac gcgtgaccaa
     1561 ctgactgaag agcagatcgc agaatttaaa gaggctttct ccctatttga caaggacggg
     1621 gatgggacga tgacaaccaa ggagctgggg acggtgatgc ggtctctggg gcagaacccc
     1681 acagaagcag agctgcagga catgatcaat gaagtagatg ccgacggtga cggcacattc
     1741 gacttccctg agttcctgac gatgatggca agaaaaatga atgacacaga cagtgaagag
     1801 gaaattagag aagcgttccg cgtgtttgat aaggacggca atggctacat cggcgcagca
     1861 gagcttcgcc acgtgatgac agaccttgga gagaagataa cagatgagga ggttgatgaa
     1921 atgatcaggg tagcagacat cgatggggat ggtcaggtaa actatgaaga gtttgtacaa
     1981 atgatgacag cgaagtagga attctgcaga tatccatcac actggcggcc gctcgagcat
     2041 gcatctagag ggccctattc tatagtgtca cctaaatgct agagctcgct gatcagcctc
     2101 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     2161 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     2221 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     2281 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     2341 aagaaccagc tggggctcta gtccggaaac tcgctgcgct cggtcgttcg gctgcggcga
     2401 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     2461 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2521 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2581 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2641 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2701 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2761 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2821 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2881 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2941 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3001 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3061 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3121 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3181 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3241 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3301 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     3361 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     3421 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     3481 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     3541 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3601 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3661 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3721 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3781 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3841 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3901 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     3961 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4021 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     4081 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     4141 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     4201 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     4261 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
     4321 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     4381 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     4441 ccgacaattg catga
//
