LOCUS       sequence-490390-        4457 bp    DNA     circular SYN 23-OCT-2025
DEFINITION  Low-affinity green GECI (cytosolic).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4457)
  AUTHORS   Davis LC, Morgan AJ, Galione A
  TITLE     Optical profiling of autonomous Ca(2+) nanodomains generated by 
            lysosomal TPC2 and TRPML1.
  JOURNAL   Cell Calcium. 2023 Dec;116:102801. doi: 10.1016/j.ceca.2023.102801. 
            Epub 2023 Sep 18.
  PUBMED    37742482
REFERENCE   2  (bases 1 to 4457)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 23, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell 
            Calcium."; date: "2023-12"; volume: "116:102801. doi"; pages: " 
            10.1016/j.ceca.2023.102801. Epub 2023 Sep 18"
COMMENT     Sequence Label: G-GECO1.2 (F90L)
FEATURES             Location/Qualifiers
     source          1..4457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(160..748)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(240..259)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(919..1710,1711..1779))
                     /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     1542..1561
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1780..1884)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1959..1978)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2150..2529
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2530..2733
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2684..2704
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2778..2797
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2778..2796
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2846..2902
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     3115..3136
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3644..4087
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREALRVFDKDGNGYIGAAEL
                     RHVMTNLGEKLTDEEVDEMIRVADIDGDGQVNYEEFVQMMTAK"
     promoter        complement(4148..4166)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4149..4166)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4186..4203)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4192..4416
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
       61 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
      121 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
      181 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
      241 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
      301 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
      361 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
      421 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
      481 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
      541 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
      601 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
      661 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
      721 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
      781 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
      841 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
      901 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
      961 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     1021 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     1081 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     1141 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     1201 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     1261 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     1321 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     1381 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     1441 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     1501 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     1561 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     1621 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     1681 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     1741 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     1801 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     1861 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     1921 atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca
     1981 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     2041 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
     2101 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
     2161 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     2221 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     2281 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     2341 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     2401 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     2461 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     2521 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     2581 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     2641 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     2701 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
     2761 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagcttg gtaccgagct
     2821 cggatccacc atggtcgact catcacgtcg taagtggaat aagacaggtc acgcagtcag
     2881 agctataggt cggctgagct cactcgagaa cgtctatatc aaggccgacg agcagaagaa
     2941 cggcatcaag gcgtacttca agatccgcca caacatcgag ggcggcggcg tgcagctcgc
     3001 ctaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     3061 ctacctgagc gtgcagtcca tgctttcgaa agaccccaac gagaagcgcg atcacatggt
     3121 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaaggg
     3181 cggtaccgga gggagcgaat ccatggtgag caagggcgag gagctgttca ccggggtggt
     3241 gcccatccag gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     3301 gggtgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     3361 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     3421 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     3481 catccaggag cgcaccatct tcttcaaggg cgacggcaac tacaagaccc gcgccgaggt
     3541 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     3601 ggacggcaac atcctggggc acaagctgga gtacaacacg cgtgaccaac tgactgaaga
     3661 gcagatcgca gaatttaaag aggctttctc cctatttgac aaggacgggg atgggacgat
     3721 aacaaccaag gagctgggga cggtgatgcg gtctctgggg cagaacccca cagaagcaga
     3781 gctgcaggac atgatcaatg aagtagatgc cgacggtgac ggcacaatcg acttccctga
     3841 gttcctgaca atgatggcaa gaaaaatgaa agacacagac agtgaagaag aaattagaga
     3901 agcgttacgt gtgtttgata aggacggcaa tggctacatc ggcgcagcag agcttcgcca
     3961 cgtgatgaca aaccttggag agaagttaac agatgaagag gttgatgaaa tgatcagggt
     4021 agcagacatc gatggggatg gtcaggtaaa ctacgaagag tttgtacaaa tgatgacagc
     4081 gaagtaagaa ttctgcagat atccatcaca ctggcggccg ctcgagcatg catctagagg
     4141 gccctattct atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt
     4201 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     4261 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     4321 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     4381 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     4441 ggggctctag tccggaa
//
