LOCUS       sequence-366181-        7424 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AKAP79-targeted Dronpa. Fluctuating green-fluorescent label for 
            live-cell super-resolution imaging..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7424)
  AUTHORS   Lin W, Mo GCH, Mehta S, Zhang J
  TITLE     DrFLINC Contextualizes Super-resolution Activity Imaging.
  JOURNAL   J Am Chem Soc. 2021 Sep 22;143(37):14951-14955. doi: 
            10.1021/jacs.1c05530. Epub 2021 Sep 13.
  PUBMED    34516108
REFERENCE   2  (bases 1 to 7424)
  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/jacs.1c05530"; journalName: "J Am Chem Soc"; date: 
            "2021-09-22- 22"; volume: "143"; issue: "37"; pages: "14951-14955"
COMMENT     Sequence Label: pcDNA3-AKAP79-Dronpa
FEATURES             Location/Qualifiers
     source          1..7424
                     /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      895..904
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             907..936
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             949..960
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2251..2925
                     /codon_start=1
                     /product="monomeric photoswitchable Dronpa-Green1 
                     fluorescent protein"
                     /label=Dronpa-Green1
                     /translation="MSVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSMDLKVK
                     EGGPLPFAYDILTTVFCYGNRVFAKYPENIVDYFKQSFPEGYSWERSMNYEDGGICNAT
                     NDITLDGDCYIYEIRFDGVNFPANGPVMQKRTVKWEPSTEKLYVRDGVLKGDVNMALSL
                     EGGGHYRCDFKTTYKAKKVVQLPDYHFVDHHIEIKSHDKDYSNVNLHEHAEAHSELPRQ
                     AK"
     CDS             2932..2958
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     promoter        complement(2977..2995)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2978..2995)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3015..3032)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3021..3245
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3291..3719
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3378..3397)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3588..3609
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3728..3748)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3733..4062
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3913..4048
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3975..3994
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4129..4923
                     /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(4183..4202)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4793..4812
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5097..5218
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5134..5153)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5188..5207
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5267..5283)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5267..5283)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5280..5302)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5291..5307
                     /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(5315..5321,5322..5339,5340..5345))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5360..5381
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5498..5515)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5669..6257)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5749..5768)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6428..7219,7220..7288))
                     /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     7051..7070
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7289..7393)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 gcttgccacc
      901 atgggccatc atcatcatca tcatcatcat catcacagca gcggccatat cgaaggtcgt
      961 catatggaaa ccacaatttc agaaattcat gtagaaaaca aggatgagaa gagatcagca
     1021 gaaggtagtc ctggggctga aaggcagaag gaaaaggcat ccatgctttg cttcaagaga
     1081 agaaagaaag cagccaaagc actgaagccc aaagctggct ctgaagctgc tgatgtggca
     1141 aggaagtgtc cacaagaagc aggagcttct gatcagccag agcccacacg gggggcctgg
     1201 gcctcactca aacgtcttgt aacacgcagg aaaaggtcag agtcttcaaa gcagcaaaag
     1261 ccattggagg gtgaaatgca acctgcaata aatgctgagg atgctgatct ttctaagaaa
     1321 aaggcaaaat ctagacttaa gattccctgc ataaaattcc caagagggcc aaaaaggagt
     1381 aatcattcca aaattataga agactcagac tgcagcatca aagtccagga agaagctgaa
     1441 attttggata tacaaacaca gaccccattg aatgatcagg caacaaaggc taagtcaacc
     1501 caggatctaa gtgaaggcat ctcacagaaa gatggtgatg aggtctgtga atcaaatgtg
     1561 agcaatagca taacttctgg agagaaagtg atttcagtag aacttggatt agataatggg
     1621 cattctgcta ttcaaacggg aactctaatc cttgaagaaa ttgaaacgat caaggaaaaa
     1681 caagatgttc aaccccagca agcaagccca cttgaaactt cagaaacaga ccatcagcag
     1741 ccagtacttt ctgatgttcc tcctttacct gcaattccag atcaacaaat tgtggaagaa
     1801 gccagtaaca gtaccctaga aagtgcacca aatggaaaag actatgaaag tacagagatt
     1861 gtagctgaag aaactaagcc aaaagatact gaattgagcc aagaatcaga ttttaaagaa
     1921 aatgggatca ctgaagagaa atccaaatca gaagaaagca aaagaatgga gccaattgct
     1981 attattatta cagacactga aatcagtgaa tttgatgtta caaaatctaa aaatgtccct
     2041 aagcaattct taatttcagc tgaaaatgag caagtagggg tttttgctaa tgataatggt
     2101 tttgaggata gaacttcaga acaatatgaa acactcttaa ttgaaacagc ctcttctcta
     2161 gtcaagaatg ctattcagtt gtcaatagaa cagctggtta atgaaatggc ctctgatgat
     2221 aataaaataa acaatcttct acaggatccg atgagtgtga ttaaaccaga catgaagatc
     2281 aagctgcgta tggaaggcgc tgtaaatgga cacccgttcg cgattgaagg agttggcctt
     2341 gggaagcctt tcgagggaaa acagagtatg gaccttaaag tcaaagaagg cggacctctg
     2401 cctttcgcct atgacatctt gacaactgtg ttctgttacg gcaacagggt attcgccaaa
     2461 tacccagaaa atatagtaga ctatttcaag cagtcgtttc ctgagggcta ctcttgggaa
     2521 cgaagcatga attacgaaga cgggggcatt tgtaacgcga caaacgacat aaccctggat
     2581 ggtgactgtt atatctatga aattcgattt gatggtgtga actttcctgc caatggtcca
     2641 gttatgcaga agaggactgt gaaatgggag ccatccactg agaaattgta tgtgcgtgat
     2701 ggagtgctga agggtgatgt taacatggct ctgtcgcttg aaggaggtgg ccattaccga
     2761 tgtgacttca aaactactta taaagctaag aaggttgtcc agttgccaga ctatcacttt
     2821 gtggaccacc acattgagat taaaagccac gacaaagatt acagtaatgt taatctgcat
     2881 gagcatgccg aagcgcattc tgagctgccg aggcaggcca agtaagaatt cctgcccccc
     2941 ctggagcgcc tgaccctgta atctagaggg ccctattcta tagtgtcacc taaatgctag
     3001 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3061 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3121 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     3181 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     3241 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     3301 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3361 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3421 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3481 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3541 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3601 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3661 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3721 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3781 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3841 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3901 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3961 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     4021 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     4081 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4141 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4201 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4261 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     4321 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4381 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4441 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4501 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4561 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4621 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4681 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4741 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4801 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4861 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4921 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     4981 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5041 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     5101 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5161 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     5221 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     5281 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5341 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5401 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5461 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5521 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5581 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5641 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5701 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5761 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5821 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5881 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5941 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6001 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6061 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6121 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6181 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6241 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6301 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6361 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6421 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6481 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6541 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6601 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6661 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6721 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6781 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6841 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6901 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6961 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7021 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7081 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7141 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7201 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7261 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7321 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7381 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
