LOCUS       sequence-382528-        6905 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Far-red fluorescent Ca2+ ion indicator for mammalian cell 
            expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6905)
  AUTHORS   Hashizume R, Fujii H, Mehta S, Ota K, Qian Y, Zhu W, Drobizhev M, 
            Nasu Y, Zhang J, Bito H, Campbell RE
  TITLE     A genetically encoded far-red fluorescent calcium ion biosensor 
            derived from a biliverdin-binding protein.
  JOURNAL   Protein Sci. 2022 Oct;31(10):e4440. doi: 10.1002/pro.4440.
  PUBMED    36173169
REFERENCE   2  (bases 1 to 6905)
  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: "Protein 
            Sci."; date: "2022-10"; volume: "31(10):e4440. doi"; pages: " 
            10.1002/pro.4440"
COMMENT     Sequence Label: pcDNA3.1(+)-iBB-GECO1
FEATURES             Location/Qualifiers
     source          1..6905
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(53..72)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        244..623
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        624..827
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     778..798
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     872..891
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        872..890
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2505..2522)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2511..2735
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2781..3209
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2868..2887)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3078..3099
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3218..3238)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3223..3552
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3403..3538
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3465..3484
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3619..4413
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3673..3692)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4283..4302
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4587..4708
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4624..4643)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4678..4697
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4757..4773)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4757..4773)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4770..4792)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4781..4797
                     /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(4805..4811,4812..4829,4830..4835))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4850..4871
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4988..5005)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5159..5747)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5239..5258)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5918..6709,6710..6778))
                     /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     6541..6560
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6779..6883)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
       61 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
      121 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
      181 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
      241 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
      301 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      361 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      421 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      481 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      541 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      601 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      661 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      721 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      781 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta
      841 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag
      901 ctggctagcg tttaaactta agcttggtac cgagctcgga tccgccacca tgttctcggt
      961 accgctgact acctcagctt tcggccacgc gtttctggct aacattgaac gtgagcagat
     1021 ccacctggcg ggctccattc agccgcacgg tatcctgctg gctgttaaag agcctgacaa
     1081 cgtggtgatc caggcttcta ttaacgctgc ggagttcctg aacaccaact ttgttgttgg
     1141 ccgtccgctg cgtgacctgg gcggcgatct gcctttgcag atcctgccgc acctgaacgg
     1201 cccgctgcac ctggctccga tgaccctgcg ttgtactgtg ggttctccgc cgcgtcgtgt
     1261 ggactgtacc attcatcgac cgtctaacgg cggcctgatc gtagaactgg aaccagcaac
     1321 caaggccact aacattgcgc cggctctggt cggtgcgctt catcgtatca cttcttcatc
     1381 ctccctgatg ggcctgtgtg acgaaaccgc gactattttc cgtgagatca ctggtttcga
     1441 ccgtgtgatg gtaatgcgtc tcggcgcgct tgacgatctg actgaagagc agatcgcaga
     1501 gattaaagag gctttctccc tatttgacaa ggacggggac gggacgataa caaccaagga
     1561 gctggggacg gtgttccggt ctctggggca gaaccccaca gaagcagagc tgcaggactt
     1621 aatcaacgag gtagatgccg atggcaacgg catattcgac ttccctgagt tcctgacgat
     1681 gatggcaagg aaaatgaatg acgcagacag tgaagaggaa attagaggag cgttccgcgt
     1741 gtttgataag gacggcaatg gcaacatcgg cgcagcaggg cttcgccacg tgatgtcaga
     1801 ccttggtgag aagttaactg atgaggaggt tgatgaaatg atcagggtag cggacaacga
     1861 tggggatggt caggtaaact acgaagagtt tgtacaaatc atgacagcga agggaggttc
     1921 tgtagattca tcacgtcgta agtggaataa ggcaggacac gcagtcagag ctataggtcg
     1981 gctgagctcg cgtaggcatg atttgctttc cgaatgtcgt cgtgcggacc tggaagcgtt
     2041 cctgggtaac cgctacccgg cgtatactat tccgcagacc gctcgtcgcc tgtacgaact
     2101 caaccgtgtt cgcctgatgg tagatgtgaa ctatactccg gttcctctag agccgcgcat
     2161 cagcccgctg aacggtcgtg atctggatat gtccctgtct tgcctgcgct ctatgtcccc
     2221 gtgccacctg aaatacatgc aggacatggg cgtgggcgca accctggttt gctctctgat
     2281 ggtgtctggt cgtctgtggg gtctgatcgt ttgccaccac tacgaaccgc gctacgttcc
     2341 gtcccacatt cgcgctgctg gcgaagcgct ggcggaagca tgtgcgaacc gcatcgcgac
     2401 gctggagagc tttgcaccgt ctcagtccaa atgagaattc tgcagatatc cagcacagtg
     2461 gcggccgctc gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     2521 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2581 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     2641 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     2701 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     2761 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2821 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2881 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2941 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3001 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3061 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3121 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3181 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     3241 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3301 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3361 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3421 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3481 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3541 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     3601 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3661 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3721 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3781 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     3841 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3901 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     3961 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4021 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4081 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4141 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4201 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4261 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4321 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4381 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     4441 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     4501 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4561 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     4621 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4681 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     4741 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     4801 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4861 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4921 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4981 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5041 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5101 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5161 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5221 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5281 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5341 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5401 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5461 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5521 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5581 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5641 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5701 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5761 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5821 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5881 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5941 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6001 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6061 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6121 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6181 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6241 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6301 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6361 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6421 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6481 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6541 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6601 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6661 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6721 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6781 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6841 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6901 tgcca
//
