LOCUS       sequence-321836-        7338 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Far-red fluorescent calcium indicator FR-GECO1c.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7338)
  AUTHORS   Dalangin R, Drobizhev M, Molina RS, Aggarwal A, Patel R, Abdelfattah
            AS, Zhao Y, Wu J, Podgorski K, Schreiter ER, Hughes TE, Campbell RE,
            Shen Y
  TITLE     Far-red fluorescent genetically encoded calcium ion indicators
  JOURNAL   bioRxiv 2020.11.12.380089
REFERENCE   2  (bases 1 to 7338)
  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: "bioRxiv 
            2020.11.12.380089"
COMMENT     Sequence Label: CAG-FR-GECO1c
FEATURES             Location/Qualifiers
     source          1..7338
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..658
                     /label=chicken beta-actin promoter
     intron          659..1675
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1599..1621
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1683..1702
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1835..1844
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3194..3211)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3200..3424
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3470..3898
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3557..3576)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3767..3788
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3907..3927)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3912..4241
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4092..4227
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4154..4173
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4252..4851
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4252..4271)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4748..4768
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5011..5132
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5048..5067)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5102..5121
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5181..5197)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5181..5197)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5194..5216)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5205..5221
                     /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(5229..5235,5236..5253,5254..5259))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5274..5295
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5412..5429)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5583..6171)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5663..5682)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6342..7133,7134..7202))
                     /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     6965..6984
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7203..7307)
                     /gene="bla"
                     /label=AmpR promoter
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 gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg ggcgcgcgcc aggsggggcg gggcggggcg aggggcgggg
      541 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
      601 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
      661 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag
      841 ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt
      901 gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg
     1021 cggtgccccg cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg
     1081 cgtggggggg tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc
     1141 ccctccccga gttgctgagc acggcccggc ttcgggtgcg gggctccgta cggggcgtgg
     1201 cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg
     1261 gccgcctcgg gccggggagg gctcggggga ggggcgcggc ggcccccgga gcgccggcgg
     1321 ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca
     1381 gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc
     1441 ctctagcggg cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg
     1501 ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc cagcctcggg gctgtccgcg
     1561 gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac
     1621 cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct
     1681 gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaattcc tcgaggaatt
     1741 cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc
     1801 tcacaaatac cactgagatc tgcatgagct agccgccacc atgggcagcg tgaagcagat
     1861 ccctagcctg accacagtga tcctggtcaa gtccatgctg cggaagcgga gcttcggcaa
     1921 ccccttcggt tttctgaccg agaccctggt gcctgtggac ggtggcctgg caggcacatg
     1981 tgacatggcc ctgagactcg ttggcggcgg acacctgaag tgcaacctga aaaccaccta
     2041 cagaagcaag aagcccgcca agaacctgaa gatgcctggc gaacaccagg tgggccggaa
     2101 gctggaaaga atcaaagagg ccgacaacga gacatacgtg gaacagcacg aagtggccgt
     2161 ggccagatat gttggacttg gcggaggcgg aggaacaggc ggatctgtgt ctgagctgat
     2221 caaagaaaac atgcggatga agctctactt ggaagggacc gtcggcaacc accacttcaa
     2281 gtgtactgcc gaaggcgagg gcaagcccta tgagggaaca cagacccagc ggatcaaggt
     2341 ggtggaaggt ggccctctgc ctttcgcctt tgacatcctg gccacctgtt ttatgtacgg
     2401 cagcaagacc ttcatcaact atccccaggg catccccgat ttcttcaagc agagcttccc
     2461 cgagggcttc acctgggaga gagtgaccac atacgaggat ggcggcgtgc tgaccgtgac
     2521 acaggataca agcctccagg acggctacct gatctacaat gtgaagctgc tgggcgtgaa
     2581 cttccccagc gatggacccg tgatgcagaa gaaaaccctc ggctgggtgc ccaccgatgg
     2641 cacgctgaca gaagaactga tcgccgagtt caaagaagcc ttcagcctgt tcgacaagga
     2701 cggcgacggc accatcacca ccaaagaact gggctccgtg atgcggagcc tgggccagaa
     2761 tccaacagaa gccgaactcc aggacatgat caacgaggtg gacgccgatg gcgacggaac
     2821 cttcgattac cctgagttcc tgaccatgat gtcccggaag atgagctaca gagtgaccga
     2881 ggaagagatc cgggaagcct tccaggtttt cgataaggat ggcaacggct acatcggagc
     2941 cgccgaactg agacgcgtga tggccgatct gggcgagaag ctgactgacg aagaggttga
     3001 cgagatgatc agagtggccg acatcgacgg cgacggacaa gtgaactacg aagagttcgt
     3061 gcagatgatg accgccaagt agaagcttgg taccgagctc ggatccacta gtccagtgtg
     3121 gtggaattct gcagatatcc agcacagtgg cggccgctcg agtctagagg gcccgtttaa
     3181 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     3241 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     3301 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     3361 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     3421 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     3481 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3541 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3601 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3661 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3721 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3781 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3841 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     3901 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3961 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     4021 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     4081 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4141 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     4201 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg catgaccgag
     4261 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
     4321 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc
     4381 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     4441 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     4501 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     4561 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     4621 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     4681 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     4741 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     4801 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg attcgaaatg
     4861 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4921 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4981 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     5041 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5101 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     5161 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     5221 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     5281 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5341 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5401 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5461 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5521 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5581 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5641 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5701 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5761 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5821 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5881 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5941 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6001 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     6061 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6121 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6181 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6241 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6301 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6361 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6421 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6481 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6541 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6601 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6661 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6721 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6781 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6841 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6901 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6961 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7021 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     7081 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7141 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7201 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7261 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7321 aaagtgccac ctgacgtc
//
