LOCUS       sequence-215031-        7230 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Genetically-encoded Cyan-Yellow Zinc FRET sensor. Localizes to the 
            cytosol. Low affinity to free zinc (Kd ~ 300nM, n = 0.55) due to 
            mutation of the zinc binding Cys residues to His..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7230)
  AUTHORS   Fiedler BL, Van Buskirk S, Carter KP, Qin Y, Carpenter MC, Palmer 
            AE, Jimenez R
  TITLE     Droplet Microfluidic Flow Cytometer For Sorting On Transient 
            Cellular Responses Of Genetically-Encoded Sensors.
  JOURNAL   Anal Chem. 2017 Jan 3;89(1):711-719. doi: 
            10.1021/acs.analchem.6b03235. Epub 2016 Dec 13.
  PUBMED    27959493
REFERENCE   2  (bases 1 to 7230)
  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"; doi: 
            "10.1021/acs.analchem.6b03235"; journalName: "Anal Chem"; date: 
            "2017-01-3- 3"; volume: "89"; issue: "1"; pages: "711-719"
COMMENT     Sequence Label: pb-NES-ZapCV5 (cpV143)
FEATURES             Location/Qualifiers
     source          1..7230
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     310..332
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     394..413
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             471..497
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             join(507..509,510..512,513..1025)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIE"
     CDS             1413..2147
                     /codon_start=1
                     /product="circularly permuted form of the Venus yellow 
                     fluorescent protein (Nagai et al., 2004)"
                     /label=cp173Venus
                     /note="efficient FRET acceptor for CFP"
                     /translation="MDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRD
                     HMVLLEFVTAAGITLGMDELYKGGSGGMVSKGEELFTGVVPILVELDGDVNGHKFSVSG
                     EGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPE
                     GYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHN
                     VYITADKQKNGIKANFKIRHNIE"
     misc_feature    2264..2813
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2408..2425)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2635..2654
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(3284..3301)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3290..3514
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3561..3577)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3561..3577)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3574..3596)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3585..3601
                     /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(3609..3615,3616..3633,3634..3639))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3654..3675
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3792..3809)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3963..4551)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4043..4062)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4722..5513,5514..5582))
                     /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     5345..5364
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5583..5687)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5713..6168)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5850..5871)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6062..6081
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6295..6317
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6309..6326
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6310..6326
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6336..6355
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6336..6354
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   6384..6418
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        6721..7100
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cggggcgcgg cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc cgcagccatt
       61 gccttttatg gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa atctgtgcgg
      121 agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg aagcggtgcg
      181 gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc cgccgtcccc
      241 ttctccctct ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg gggggacggg
      301 gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagagcctct gctaaccatg
      361 ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt gtgctgtctc
      421 atcattttgg caaagaattc aacttaagct tggtaccgag catgcttcaa cttcctcctc
      481 ttgaacgtct tactctttcg gatccgatgg tgagcaaggg cgaggagctg ttcaccgggg
      541 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaggttc agcgtgtccg
      601 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
      661 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctggggc gtgcagtgct
      721 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
      781 gctacgtcca ggagcgtacc atcttcttca aggacgacgg caactacaag acccgcgccg
      841 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
      901 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacatcagc cacaacgtct
      961 atatcaccgc cgacaagcag aagaacggca tcaaggccca cttcaagatc cgccacaaca
     1021 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     1081 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     1141 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc cgcatgcata
     1201 aaaacaatga cttaaaacac aaatggaaag aacatcctga gtcttgtagc tcactatttg
     1261 acctacaaag acatcttttg aaggatcatg tctctcaaga tttcaaacac cctatggaac
     1321 cattagctca taattgggag gaccatgatt tccttggaga tgatacatgt tccatagtga
     1381 accatattaa ttgtcaacat ggtatcgagc tcatggacgg cggcgtgcag ctcgccgacc
     1441 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     1501 tgagctacca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     1561 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggtggca
     1621 gcggtggcat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     1681 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     1741 ccacctacgg caagctgacc ctgaagctga tctgcaccac cggcaagctg cccgtgccct
     1801 ggcccaccct cgtgaccacc ctgggctacg gcctgcagtg cttcgcccgc taccccgacc
     1861 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     1921 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     1981 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     2041 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcacc gccgacaagc
     2101 agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgagtaa gaatcctgca
     2161 gatatccagc acagtgggtc gacgggccgc ggtaacaatt gttaactaac ttaagctagc
     2221 aacggtttcc ctctagcggg atcaattccg cccccccccc ctaacgttac tggccgaagc
     2281 cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct
     2341 tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt
     2401 ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct
     2461 ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc
     2521 ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag
     2581 gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc
     2641 tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga
     2701 tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt
     2761 ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgata ataccatggt
     2821 catgaaaaca tttaacattt ctcaacaaga tctagaatta gtagaagtag cgacagagaa
     2881 gattacaatg ctttatgagg ataataaaca tcatgtggga gcggcaattc gtacgaaaac
     2941 aggagaaatc atttcggcag tacatattga agcgtatata ggacgagtaa ctgtttgtgc
     3001 agaagccatt gcgattggta gtgcagtttc gaatggacaa aaggattttg acacgattgt
     3061 agctgttaga cacccttatt ctgacgaagt agatagaagt attcgagtgg taagtccttg
     3121 tggtatgtgt agggagttga tttcagacta tgcaccagat tgttttgtgt taatagaaat
     3181 gaatggcaag ttagtcaaaa ctacgattga agaactcatt ccactcaaat atacccgaaa
     3241 ttaaaagtcg agatgcatcg atgatctaga gctcgctgat cagcctcgac tgtgccttct
     3301 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     3361 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3421 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3481 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aacctgcagc
     3541 ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     3601 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     3661 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     3721 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3781 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3841 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3901 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3961 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4021 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4081 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4141 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4201 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4261 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4321 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4381 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4441 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4501 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4561 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4621 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4681 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4741 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4801 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4861 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4921 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4981 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5041 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5101 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5161 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5221 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5281 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5341 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5401 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5461 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5521 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5581 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5641 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5701 aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     5761 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     5821 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     5881 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     5941 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     6001 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     6061 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     6121 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat
     6181 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     6241 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     6301 cacgacgttg taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat
     6361 tggggcgcgc cattctagat taaccctaga aagatagtct gcgtaaaatt gacgcatgca
     6421 ttcttgaaat attgctctct ctttctaaat agcgcgaatc cgtcgctgtg catttaggac
     6481 atctcagtcg ccgcttggag ctcccgtgag gcgtgcttgt caatgcggta agtgtcactg
     6541 attttgaact ataacgaccg cgtgagtcaa aatgacgcat gattatcttt tacgtgactt
     6601 ttaagattta actcatacga taattatatt gttatttcat gttctactta cgtgataact
     6661 tattatatat atattttctt gttatagata tcaactagaa tgctagcatg ggcccatctc
     6721 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     6781 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     6841 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     6901 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     6961 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     7021 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     7081 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
     7141 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
     7201 tgggggcggg gggggggggg gggcgcgcgc
//
