LOCUS       sequence-363422-        8024 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PKA RII-alpha subunit-targeted Dronpa chromophore-removed FLINC 
            (DrFLINC) pair. Fluctuating red-fluorescent label for live-cell 
            super-resolution imaging..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8024)
  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 8024)
  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-RII-alpha-uDpTT
FEATURES             Location/Qualifiers
     source          1..8024
                     /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"
     CDS             2125..2796
                     /codon_start=1
                     /product="monomeric photoswitchable Dronpa-Green1 
                     fluorescent protein"
                     /label=Dronpa-Green1
                     /translation="MSVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSMDLKVK
                     EGGPLPFAYDILTTVFCYGNRVFAKYPENIVDYFKQSFPEGYSWERSMNYEDGGICNAT
                     NDITLDGDCYIYEIRFDGVNFPANGPVMQKRTVKWEPSTEKLYVRDGVLKGDVNMALSL
                     EGGGHYRCDFKTTYKAKKVVQLPDYHFVDTHIEIKSHDKDYSNVNLHEHAEAHSELPRQ
                     AK"
     CDS             2821..3516
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTDMALK
                     LVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHK"
     promoter        complement(3577..3595)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3578..3595)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3615..3632)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3621..3845
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3891..4319
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3978..3997)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4188..4209
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4328..4348)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4333..4662
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4513..4648
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4575..4594
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4729..5523
                     /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(4783..4802)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5393..5412
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5697..5818
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5734..5753)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5788..5807
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5867..5883)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5867..5883)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5880..5902)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5891..5907
                     /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(5915..5921,5922..5939,5940..5945))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5960..5981
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6098..6115)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6269..6857)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6349..6368)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7028..7819,7820..7888))
                     /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     7651..7670
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7889..7993)
                     /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 gcttgccgcc
      901 accatgagcc acatccagat cccgccgggg ctcacggagc tgctgcaggg ctacacggtg
      961 gaggtgctgc gacagcagcc gcctgacctc gtcgaattcg cagtggagta cttcacccgc
     1021 ctgcgcgagg cccgcgcccc agcctcagtc ctgcccgccg ccaccccacg ccagagcctg
     1081 ggccaccccc cgccagaacc cggcccggac cgtgtcgccg acgccaaagg ggacagcgag
     1141 tcggaggagg acgaggactt ggaagttcca gttcctagca gatttaatag acgagtatca
     1201 gtctgtgctg agacctataa ccctgatgag gaagaggaag atacagatcc aagggtgatt
     1261 catcctaaaa ctgatgaaca gagatgcaga cttcaggaag cttgcaaaga tattctcctt
     1321 ttcaaaaatc ttgatcagga acagctttct caagttctcg atgccatgtt tgaaaggata
     1381 gtcaaagctg atgagcatgt cattgaccaa ggagatgatg gagacaactt ttatgtcata
     1441 gaacggggaa cttatgacat tttagtaaca aaagataatc aaacccgctc tgttggtcaa
     1501 tatgacaacc gtggcagttt tggagaacta gctctgatgt acaacacccc gagagctgct
     1561 accattgttg ctacctcaga aggctccctt tggggactgg accgggtgac ttttagaaga
     1621 atcatagtga aaaataatgc aaagaagagg aagatgtttg aatcatttat tgagtctgtg
     1681 cccctcctta aatcactaga ggtgtcagaa cgaatgaaga ttgtggatgt aataggagag
     1741 aagatctata aggatggaga acgcataatc actcagggtg aaaaggctga tagcttttac
     1801 atcatagagt ctggcgaagt gagcatcttg attagaagca ggactaaatc aaacaaggat
     1861 ggtgggaacc aggaggtcga gattgcccgc tgccataagg ggcagtactt tggagagctt
     1921 gccctggtca ccaacaaacc cagagctgcc tcagcttatg cagttggaga tgtcaaatgc
     1981 ttagttatgg atgtacaagc attcgagagg cttctggggc cctgcatgga catcatgaag
     2041 aggaacatct cacactatga ggaacagctg gtgaagatgt ttggctccag cgtggatctg
     2101 ggcaacctcg ggcaggggga tcccatgagt gtgattaaac cagacatgaa gatcaagctg
     2161 cgtatggaag gcgctgtaaa tggacacccg ttcgcgattg aaggagttgg ccttgggaag
     2221 cctttcgagg gaaaacagag tatggacctt aaagtcaaag aaggcggacc tctgcctttc
     2281 gcctatgaca tcttgacaac tgtgttctgt tacggcaaca gggtattcgc caaataccca
     2341 gaaaatatag tagactattt caagcagtcg tttcctgagg gctactcttg ggaacgaagc
     2401 atgaattacg aagacggggg catttgtaac gcgacaaacg acataaccct ggatggtgac
     2461 tgttatatct atgaaattcg atttgatggt gtgaactttc ctgccaatgg tccagttatg
     2521 cagaagagga ctgtgaaatg ggagccatcc actgagaaat tgtatgtgcg tgatggagtg
     2581 ctgaagggtg atgttaacat ggctctgtcg cttgaaggag gtggccatta ccgatgtgac
     2641 ttcaaaacta cttataaagc taagaaggtt gtccagttgc cagactatca ctttgtggac
     2701 acacacattg agattaaaag ccacgacaaa gattacagta atgttaatct gcatgagcat
     2761 gccgaagcgc attctgagct gccgaggcag gccaagagat ctgacgaggt ggacggtacc
     2821 atgagcgagc tgattaagga gaacatgcac atgaagctgt acatggaggg caccgtgaac
     2881 aaccaccact tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc
     2941 atgagaatca aggtggtcga gggcggccct ctccccttcg ccttcgacat cctggctacc
     3001 agcttcatgt acggcagcag aaccttcatc aaccacaccc agggcatccc cgatttcttt
     3061 aagcagtcct tccctgaggg cttcacatgg gagagagtca ccacatacga agacgggggc
     3121 gtgctgaccg ctacccagga caccagcctc caggacggct gcctcatcta caacgtcaag
     3181 atcagagggg tgaacttccc atccaacggc cctgtgatgc agaagaaaac actcggctgg
     3241 gaggccaaca ccgagatgct gtaccccgct gacggcggcc tggaaggcag aaccgacatg
     3301 gccctgaagc tcgtgggcgg gggccacctg atctgcaact tcaagaccac atacagatcc
     3361 aagaaacccg ctaagaacct caagatgccc ggcgtctact atgtggacca cagactggaa
     3421 agaatcaagg aggccgacaa agagacctac gtcgagcagc acgaggtggc tgtggccaga
     3481 tactgcgacc tccctagcaa actggggcac aagtaagaat tctgcagata tccatcacac
     3541 tggcggccgc tcgagcatgc atctagaggg ccctattcta tagtgtcacc taaatgctag
     3601 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3661 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3721 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     3781 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     3841 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     3901 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3961 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4021 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4081 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     4141 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4201 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4261 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     4321 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4381 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4441 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4501 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4561 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     4621 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     4681 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4741 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4801 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4861 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     4921 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4981 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     5041 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     5101 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     5161 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     5221 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     5281 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5341 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5401 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5461 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5521 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     5581 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5641 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     5701 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5761 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     5821 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     5881 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5941 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6001 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     6061 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6121 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6181 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6241 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6301 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6361 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6421 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6481 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6541 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6601 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6661 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6721 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6781 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6841 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6901 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6961 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7021 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7081 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7141 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7201 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7261 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7321 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7381 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7441 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7501 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7561 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7621 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7681 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7741 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7801 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7861 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7921 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7981 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
