LOCUS       sequence-321835-        7330 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Far-red fluorescent calcium indicator FR-GECO1a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7330)
  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 7330)
  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-GECO1a
FEATURES             Location/Qualifiers
     source          1..7330
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          651..1667
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1591..1613
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1675..1694
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1827..1836
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3186..3203)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3192..3416
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3462..3890
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3549..3568)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3759..3780
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3899..3919)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3904..4233
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4084..4219
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4146..4165
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4244..4843
                     /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(4244..4263)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4740..4760
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5003..5124
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5040..5059)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5094..5113
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5173..5189)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5173..5189)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5186..5208)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5197..5213
                     /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(5221..5227,5228..5245,5246..5251))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5266..5287
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5404..5421)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5575..6163)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5655..5674)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6334..7125,7126..7194))
                     /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     6957..6976
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7195..7299)
                     /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 ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg ggcggggcga
      541 ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg
      601 cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg
      661 cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct
      721 gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta
      781 attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg
      841 ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt
      901 gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg
      961 gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc
     1021 cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg
     1081 ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc
     1141 gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc
     1201 tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc
     1261 gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag
     1321 gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc
     1381 ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg
     1441 ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg
     1501 cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg
     1561 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
     1621 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
     1681 tgctggttat tgtgctgtct catcattttg gcaaagaatt cctcgaggaa ttcactcctc
     1741 aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg gctcacaaat
     1801 accactgaga tctgcatgag ctagccgcca ccatgggcag cgtgaagcag atccctagcc
     1861 tgaccacagt gatcctggtc aagtccatgc tgcggaagcg gagcttcggc aaccccttcg
     1921 gttttctgac cgagtacctg gtgcctgtgg acggtggcct ggcaggcaca tgtgacatgg
     1981 ccctgagact cgttggcggc ggacacctga agtgcaacct gaaaaccacc tacagaagca
     2041 agaagcccgc caagaacctg aagatgcctg gcgaacacca ggtgggccgg aagctggaaa
     2101 gaatcaaaga ggccgacaac gagacatacg tggaacagca cgaagtggcc gtggccagat
     2161 atgttggact tggcggaggc ggaggaacag gcggatctgt gtctgagctg atcaaagaaa
     2221 acatgcggat gaagctctac ttggaaggga ccgtcggcaa ccaccacttc aagtgtactg
     2281 ccgaaggcga gggcaagccc tatgagggaa cacagaccca gcggatcaag gtggtggaag
     2341 gtggccctct gcctttcgcc tttgacatcc tggccacctg ttttatgtac ggcagcaaga
     2401 ccttcatcaa ctatccccag ggcatccccg atttcttcaa gcagagcttc cccgagggct
     2461 tcacctggga gagagtgacc acatacgagg atggcggcgt gctgaccgtg acacaggata
     2521 caagcctcca ggacggctac ctgatctaca atgtgaagct gctgggcgtg aacttcccca
     2581 gcgatggacc cgtgatgcag aagaaaaccc tcggctgggt gcccaccgat ggcacgctga
     2641 cagaagaact gatcgccgag ttcaaagaag ccttcagcct gttcgacaag gacggcgacg
     2701 gcaccatcac caccaaagaa ctgggctccg tgatgcggag cctgggccag aatccaacag
     2761 aagccgaact ccaggacatg atcaacgagg tggacgccga tggcgacgga accttcgatt
     2821 accctgagtt cctgaccatg atgtcccgga agatgagcta cagagtgacc gaggaagaga
     2881 tccgggaagc cttccaggtt ttcgataagg atggcaacgg ctacatcgga gccgccgaac
     2941 tgagacgcgt gatggccgat ctgggcgaga agctgactga cgaagaggtt gacgagatga
     3001 tcagagtggc cgacatcgac ggcgacggac aagtgaacta cgaagagttc gtgcagatga
     3061 tgaccgccaa gtagaagctt ggtaccgagc tcggatccac tagtccagtg tggtggaatt
     3121 ctgcagatat ccagcacagt ggcggccgct cgagtctaga gggcccgttt aaacccgctg
     3181 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3241 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3301 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     3361 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     3421 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     3481 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3541 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3601 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3661 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3721 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3781 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     3841 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     3901 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3961 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4021 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4081 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4141 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     4201 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgcatgaccg agtacaagcc
     4261 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc
     4321 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt
     4381 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     4441 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     4501 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     4561 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     4621 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     4681 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     4741 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     4801 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgattcgaaa tgaccgacca
     4861 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     4921 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     4981 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5041 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5101 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5161 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5221 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5281 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5341 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5401 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5461 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5521 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5581 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5641 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5701 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5761 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5821 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5881 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5941 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6001 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6061 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6121 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6181 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6241 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6301 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6361 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6421 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6481 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6541 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6601 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6661 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6721 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6781 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6841 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6901 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6961 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7021 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7081 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7141 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7201 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7261 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7321 acctgacgtc
//
