LOCUS       sequence-39588-e        8052 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8052)
  AUTHORS   Ding Y, Ai HW, Hoi H, Campbell RE
  TITLE     Forster resonance energy transfer-based biosensors for 
            multiparameter ratiometric imaging of Ca2+ dynamics and caspase-3 
            activity in single cells.
  JOURNAL   Anal Chem. 2011 Dec 15;83(24):9687-93. doi: 10.1021/ac202595g. Epub 
            2011 Nov 28.
  PUBMED    22080726
REFERENCE   2  (bases 1 to 8052)
  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/ac202595g";
            journalName: "Anal Chem"; date: "2011-12-15- 15"; volume: "83"; 
            issue: "24"; pages: "9687-93"
COMMENT     Sequence Label: CaYin1 NES
FEATURES             Location/Qualifiers
     source          1..8052
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      898..907
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2857..2878)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(3604..3622)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3605..3622)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3642..3659)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3648..3872
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3918..4346
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4005..4024)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4215..4236
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4355..4375)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4360..4690
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4541..4676
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4603..4622
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4757..5551
                     /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(4811..4830)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5421..5440
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5725..5846
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5762..5781)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5816..5835
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5895..5911)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5895..5911)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5908..5930)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5919..5935
                     /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(5943..5949,5950..5967,5968..5973))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5988..6009
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6126..6143)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6297..6885)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6377..6396)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7056..7847,7848..7916))
                     /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     7679..7698
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7917..8021)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctagccgcc
      901 accatggcct cgagcatggt gagcaagggc gaggaggtca tcaaagagtt catgcgcttc
      961 aaggtgcgca tggagggctc catgaacggc cacgagttcg agatcgaggg cgagggcgag
     1021 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg cggccccctg
     1081 cccttcgcct gggacatcct gtccccccag ttcatgtacg gctccaaggc gtacgtgaag
     1141 caccccgccg acatccccga ttacaagaag ctgtccttcc ccgagggctt caagtgggag
     1201 cgcgtgatga acttcgagga cggcggtctg gtgaccgtga cccaggactc ctccctgcag
     1261 gacggcacgc tgatctacaa ggtgaagatg cgcggcacca acttcccccc cgacggcccc
     1321 gtaatgcaga agaagaccat gggctgggag gcctccaccg agcgcctgta cccccgcgac
     1381 ggcgtgctga agggcgagat ccaccaggcc ctgaagctga aggacggcgg ccactacctg
     1441 gtggagttca agaccatcta catggccaag aagcccgtgc aactgcccgg ctactactac
     1501 gtggacacca agctggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     1561 gagcgctccg agggccgcca ccacctgttc ctggggcatg gcaccggcag caccggcagc
     1621 ggcagctccg gcaccgcctc ctccgaggac aacaacatgg ccgtcatcaa agagttcatg
     1681 cgcttcaagg tgcgcatgga gggctccatg aacggccacg agttcgagat cgagggcgag
     1741 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc
     1801 cccctgccct tcgcctggga catcctgtcc ccccagttca tgtacggctc caaggcgtac
     1861 gtgaagcacc ccgccgacat ccccgattac aagaagctgt ccttccccga gggcttcaag
     1921 tgggagcgcg tgatgaactt cgaggacggc ggtctggtga ccgtgaccca ggactcctcc
     1981 ctgcaggacg gcacgctgat ctacaaggtg aagatgcgcg gcaccaactt cccccccgac
     2041 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg cctgtacccc
     2101 cgcgacggcg tgctgaaggg cgagatccac caggccctga agctgaagga cggcggccac
     2161 tacctggtgg agttcaagac catctacatg gccaagaagc ccgtgcaact gcccggctac
     2221 tactacgtgg acaccaagct ggacatcacc tcccacaacg aggactacac catcgtggaa
     2281 cagtacgagc gctccgaggg ccgccaccac ctgttcctgc gcatgcatct gacagaagag
     2341 cagattgcag agttcaaaga agccttctca ttattggaca aggatgggga cggcaccatc
     2401 accacaaagg aacttggcac cgctctgagg tcgcttggac aaaacccaac ggaagcagaa
     2461 ttgcaggata tgatcaatga agtcgatgct gatggcaatg gaacgattta ctttcctgaa
     2521 tttcttacta tgatggctag aaaaatgaag gacacagaca gcgaagagga aatccgagaa
     2581 gcattccgtg tttttgacaa ggatgggaac ggctacatca gcgctgctga attacgtcac
     2641 gtcatgacaa acctcgggga gaagttaaca gatgaagaag ttgatgaaat gataagggaa
     2701 gcagatatcg atggtgatgg ccaagtaaac tatgaagagt ttgtacaaat gatgacagca
     2761 aaggggggga agaggcgctg gcagaaaaca ggccatgctg tccgagcatt cggaagactg
     2821 aagaagatcg agctcttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     2881 aacggccaca ggttcagcgt gcgcggcgag ggcgagggcg atgccaccaa cggcaagctg
     2941 atcctgaagt tcatctgcac ctccggcgag ctgcccgtgc cctggcccac cctcgtgacc
     3001 accctgtctt acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
     3061 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     3121 gacggcagct acaggacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     3181 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     3241 tacaacatga acgcgtggga cgcgtatatc acggccgaca agcagaagaa cggcatcaaa
     3301 gcgaacttca agatcgagca caacgtcaag gacggcggcg tgcagctcgc cgacgcgtac
     3361 cagcagaaca cccccatcgg cggcggcccc gtgctgctgc ctgacaacca ctacctgagc
     3421 ttccagagca agctgttcaa agacccctac gagcagcgcg atcacatggt cctgctggag
     3481 ttcgttaccg ccgccgggat aactcccggc atggacgagc tgtacaagtt agccttgaaa
     3541 ttagcaggtc ttgatatcgg gagctaagaa ttcatgggca ggccggccta aactgggccc
     3601 tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg tgccttctag
     3661 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     3721 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     3781 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     3841 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     3901 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3961 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4021 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc
     4081 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4141 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4201 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4261 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct
     4321 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     4381 aagtccccag gctccccagg caggcagaag tatgcaaagc atgcatctca attagtcagc
     4441 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4501 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4561 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     4621 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4681 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     4741 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     4801 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     4861 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     4921 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     4981 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     5041 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     5101 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     5161 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     5221 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     5281 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     5341 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     5401 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     5461 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     5521 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     5581 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     5641 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     5701 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     5761 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5821 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     5881 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5941 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     6001 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     6061 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     6121 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     6181 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     6241 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     6301 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6361 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6421 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6481 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6541 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6601 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6661 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6721 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6781 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6841 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6901 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6961 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     7021 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     7081 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     7141 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     7201 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     7261 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     7321 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     7381 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     7441 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7501 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7561 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7621 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7681 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7741 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7801 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7861 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7921 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7981 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     8041 ccacctgacg tc
//
