LOCUS       sequence-145563-        8098 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FPX biosensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8098)
  AUTHORS   Ding Y, Li J, Enterina JR, Shen Y, Zhang I, Tewson PH, Mo GC, Zhang 
            J, Quinn AM, Hughes TE, Maysinger D, Alford SC, Zhang Y, Campbell RE
  TITLE     Ratiometric biosensors based on dimerization-dependent fluorescent 
            protein exchange.
  JOURNAL   Nat Methods. 2015 Jan 26. doi: 10.1038/nmeth.3261.
  PUBMED    25622108
REFERENCE   2  (bases 1 to 8098)
  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: "Nat 
            Methods. 2015 Jan 26. doi: 10.1038/nmeth.3261."
COMMENT     Sequence Label: single polypeptide FPX biosensor for calcium ion
FEATURES             Location/Qualifiers
     source          1..8098
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(63..80)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(234..822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(314..333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(993..1784,1785..1853))
                     /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     1616..1635
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1854..1958)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2033..2052)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2224..2603
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2604..2807
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2758..2778
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2852..2871
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2852..2870
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2895..2904
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5678..5695)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5684..5908
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5954..6382
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6041..6060)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6251..6272
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6391..6411)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6396..6725
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6576..6711
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6638..6657
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6792..7586
                     /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(6846..6865)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7456..7475
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7760..7881
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7797..7816)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7851..7870
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7930..7946)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7930..7946)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7943..7965)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7954..7970
                     /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(7978..7984,7985..8002,8003..8008))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8023..8044
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
       61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      421 ctttctcata gctcacgcta taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     1861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1981 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     2041 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     2101 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     2161 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
     2221 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     2281 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     2341 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     2401 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     2461 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     2521 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     2581 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
     2641 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     2701 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     2761 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta
     2821 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag
     2881 ctggctagcg tgccgccacc atggcctcga gaatggtgag caagagcgag gaggtcatca
     2941 aagagttcat gcgcttcaag gtgcgcttgg agggctccat gaacggccac gagttcgaga
     3001 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
     3061 ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc cccccagatc atgtacggct
     3121 ccaaggcgta cgtgaagcac cccgccgacg tccccgatta catgaagctg tccttccccg
     3181 agggcttcaa gtgggagcgc gtgatgcact tcgaggacgg cggtctggtg accgtgactc
     3241 aggacacctc cctgcaggac ggcacgctga tctacaaggt gaagatgcgc ggcaccaact
     3301 tcccccccga cggccccgta atgcagaaga agaccttggg ctgggattat gccaccgagc
     3361 gcctgtaccc cgaagacggc gtgctgaagg gcgagcttct ggggcgcctg aagctgaagg
     3421 acggcggcct caacctggtg gagttcaaga ccatctacat ggccaagaag cccgtgcaac
     3481 tgcccggcta ctacttcgtg gacaccaagc tgggcatcac ctcccacaac gaggactaca
     3541 ccatcgtgga acagtacgag cgctccgagg gccgccacca cctggggcat ggcaccggca
     3601 gcaccggcag cggcagctcc ggtaccgcct cctccgagga caacaacatg gccgaccaac
     3661 tgacagaaga gcagattgca gagttcaaag aagccttctc attattggac aaggatgggg
     3721 acggcaccat caccacaaag gaacttggca ccgctctgag gtcgcttgga caaaacccaa
     3781 cggaagcaga attgcaggat atgatcaatg aagtcgatgc tgatggcaat ggaacgattt
     3841 actttcctga atttcttact atgatggcta gaaaaatgaa ggacacagac agcgaagagg
     3901 aaatccgaga agcattccgt gtttttgaca aggatgggaa cggctacatc agcgctgctg
     3961 aattacgtca cgtcatgaca aacctcgggg agaagttaac agatgaagaa gttgatgaaa
     4021 tgataaggga agcagatatc gatggtgatg gccaagtaaa ctatgaagag tttgtacaaa
     4081 tgatgacagc aaagggaagc ggtggatcca tggtgagcaa gagcgaggag accatcaaag
     4141 agttcatgcg cttcaaggtg cgcatggagg gctccatgaa cggccacgag ttcgagatcg
     4201 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
     4261 agggcggccc cctgcccttc gcctgggaca tcctgtcccc ccagttcatg tacggctccg
     4321 aggcgtacgt gaggcacccc gccgacatcc ccgattacaa gaagctgccc ttccccgagg
     4381 gcttcaagtg ggagcgcgtg atgaacttcg aagacggcgg tctggtgacc gttacccagg
     4441 actcctccct gcaggacggc acgctgatct gcaaggtgaa gatgcgcggc accaacttcc
     4501 cccccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc accgagatgc
     4561 tgtaccccga agacggcgtg ctgaagggcc atagctatca ggccctgaag ctgaaggacg
     4621 gcggccacta cctggtggag ttcgagacca tctacatggc caagaagccc gtgcaactgc
     4681 ccggcgatta ctgtgtggac accaagctgg acatcacctc ccacaacgag gactacacca
     4741 tcgtggaaca gtacgggcgc tccgagggcc gccaccgtct gggcatggac gagcggtaca
     4801 agagcacgga attcgccacc atgaagaggc gctggcagaa aacaggccat gctgtccgag
     4861 cattcggaag actgaagaag atctccagct ccggggcact ggggcatggc accggcagca
     4921 ccggcagcgg cagctccggt accatggtga gcaagagcga ggaggtcatc aaagagttca
     4981 tgcgcttcaa ggtgcgcttg gagggctcca tgaacggcca cgagttcgag atcgagggcg
     5041 agggcgaggg ccgcccttac gagggcaccc agaccgccaa gctgaaggtg accaagggcg
     5101 gccccctgcc cttcgcctgg gacatcctgt cccccctgat catgtacggc tccaagatgt
     5161 acgtgaagca ccccgccgac gtccccgatt acatgaagct gtccttcccc gagggcttca
     5221 agtgggagcg cgtgatgcac ttcgaggacg gcggtctggt gaccgcgaca caggactcct
     5281 ccctgcagga cggcacgctg atctacaagg tgaagatgcg cggcaccaac ttcccccccg
     5341 acggccccgt aatgcagaag aagaccttgg gctgggatta tgccaccgag cgcctgtacc
     5401 ccgaagaagg cgtgctgaag ggcgagctcc tggggcgcct gaagctgaag gacggcggcc
     5461 tcaacctggt ggagtccaag accatctaca tggccaagaa gcccgtgcaa ctgcccggct
     5521 actacttcgt ggacaccaag ctggacatca cctcccacaa cgaggactac accatcgtgg
     5581 aacagtacga gcgctccgag ggccgccacc acctggggca tggtacaggt agcacaggca
     5641 gcggtagctc ctaaaagctt ggtaaaccgc tgatcagcct cgactgtgcc ttctagttgc
     5701 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     5761 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     5821 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     5881 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     5941 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     6001 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     6061 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     6121 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     6181 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     6241 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     6301 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     6361 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     6421 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     6481 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     6541 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     6601 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     6661 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     6721 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     6781 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     6841 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     6901 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     6961 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     7021 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     7081 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     7141 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     7201 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     7261 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     7321 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     7381 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     7441 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     7501 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     7561 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     7621 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     7681 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     7741 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     7801 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     7861 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     7921 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     7981 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     8041 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctg
//
