LOCUS       sequence-366176-        8051 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Beta-2 adrenergic receptor-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 8051)
  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 8051)
  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-beta-2AR-uDpTT
FEATURES             Location/Qualifiers
     source          1..8051
                     /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"
     CDS             2152..2823
                     /codon_start=1
                     /product="monomeric photoswitchable Dronpa-Green1 
                     fluorescent protein"
                     /label=Dronpa-Green1
                     /translation="MSVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSMDLKVK
                     EGGPLPFAYDILTTVFCYGNRVFAKYPENIVDYFKQSFPEGYSWERSMNYEDGGICNAT
                     NDITLDGDCYIYEIRFDGVNFPANGPVMQKRTVKWEPSTEKLYVRDGVLKGDVNMALSL
                     EGGGHYRCDFKTTYKAKKVVQLPDYHFVDTHIEIKSHDKDYSNVNLHEHAEAHSELPRQ
                     AK"
     CDS             2848..3543
                     /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(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..4689
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4540..4675
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4602..4621
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4756..5550
                     /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(4810..4829)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5420..5439
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5724..5845
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5761..5780)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5815..5834
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5894..5910)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5894..5910)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5907..5929)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5918..5934
                     /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(5942..5948,5949..5966,5967..5972))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5987..6008
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6125..6142)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6296..6884)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6376..6395)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7055..7846,7847..7915))
                     /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     7678..7697
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7916..8020)
                     /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 accatggggc aacccgggaa cggcagcgcc ttcttgctgg cacccaatag aagccatgcg
      961 ccggaccacg acgtcacgca gcagagggac gaggtgtggg tggtgggcat gggcatcgtc
     1021 atgtctctca tcgtcctggc catcgtgttt ggcaatgtgc tggtcatcac agccattgcc
     1081 aagttcgagc gtctgcagac ggtcaccaac tacttcatca cttcactggc ctgtgctgat
     1141 ctggtcatgg gcctggcagt ggtgcccttt ggggccgccc atattcttat gaaaatgtgg
     1201 acttttggca acttctggtg cgagttttgg acttccattg atgtgctgtg cgtcacggcc
     1261 agcattgaga ccctgtgcgt gatcgcagtg gatcgctact ttgccattac ttcacctttc
     1321 aagtaccaga gcctgctgac caagaataag gcccgggtga tcattctgat ggtgtggatt
     1381 gtgtcaggcc ttacctcctt cttgcccatt cagatgcact ggtaccgggc cacccaccag
     1441 gaagccatca actgctatgc caatgagacc tgctgtgact tcttcacgaa ccaagcctat
     1501 gccattgcct cttccatcgt gtccttctac gttcccctgg tgatcatggt cttcgtctac
     1561 tccagggtct ttcaggaggc caaaaggcag ctccagaaga ttgacaaatc tgagggccgc
     1621 ttccatgtcc agaaccttag ccaggtggag caggatgggc ggacggggca tggactccgc
     1681 agatcttcca agttctgctt gaaggagcac aaagccctca agacgttagg catcatcatg
     1741 ggcactttca ccctctgctg gctgcccttc ttcatcgtta acattgtgca tgtgatccag
     1801 gataacctca tccgtaagga agtttacatc ctcctaaatt ggataggcta tgtcaattct
     1861 ggtttcaatc cccttatcta ctgccggagc ccagatttca ggattgcctt ccaggagctt
     1921 ctgtgcctgc gcaggtcttc tttgaaggcc tatgggaatg gctactccag caacggcaac
     1981 acaggggagc agagtggata tcacgtggaa caggagaaag aaaataaact gctgtgtgaa
     2041 gacctcccag gcacggaaga ctttgtgggc catcaaggta ctgtgcctag cgataacatt
     2101 gattcacaag ggaggaattg tagtacaaat gactcactgc tgggggatcc aatgagtgtg
     2161 attaaaccag acatgaagat caagctgcgt atggaaggcg ctgtaaatgg acacccgttc
     2221 gcgattgaag gagttggcct tgggaagcct ttcgagggaa aacagagtat ggaccttaaa
     2281 gtcaaagaag gcggacctct gcctttcgcc tatgacatct tgacaactgt gttctgttac
     2341 ggcaacaggg tattcgccaa atacccagaa aatatagtag actatttcaa gcagtcgttt
     2401 cctgagggct actcttggga acgaagcatg aattacgaag acgggggcat ttgtaacgcg
     2461 acaaacgaca taaccctgga tggtgactgt tatatctatg aaattcgatt tgatggtgtg
     2521 aactttcctg ccaatggtcc agttatgcag aagaggactg tgaaatggga gccatccact
     2581 gagaaattgt atgtgcgtga tggagtgctg aagggtgatg ttaacatggc tctgtcgctt
     2641 gaaggaggtg gccattaccg atgtgacttc aaaactactt ataaagctaa gaaggttgtc
     2701 cagttgccag actatcactt tgtggacaca cacattgaga ttaaaagcca cgacaaagat
     2761 tacagtaatg ttaatctgca tgagcatgcc gaagcgcatt ctgagctgcc gaggcaggcc
     2821 aagagatctg acgaggtgga cggtaccatg agcgagctga ttaaggagaa catgcacatg
     2881 aagctgtaca tggagggcac cgtgaacaac caccacttca agtgcacatc cgagggcgaa
     2941 ggcaagccct acgagggcac ccagaccatg agaatcaagg tggtcgaggg cggccctctc
     3001 cccttcgcct tcgacatcct ggctaccagc ttcatgtacg gcagcagaac cttcatcaac
     3061 cacacccagg gcatccccga tttctttaag cagtccttcc ctgagggctt cacatgggag
     3121 agagtcacca catacgaaga cgggggcgtg ctgaccgcta cccaggacac cagcctccag
     3181 gacggctgcc tcatctacaa cgtcaagatc agaggggtga acttcccatc caacggccct
     3241 gtgatgcaga agaaaacact cggctgggag gccaacaccg agatgctgta ccccgctgac
     3301 ggcggcctgg aaggcagaac cgacatggcc ctgaagctcg tgggcggggg ccacctgatc
     3361 tgcaacttca agaccacata cagatccaag aaacccgcta agaacctcaa gatgcccggc
     3421 gtctactatg tggaccacag actggaaaga atcaaggagg ccgacaaaga gacctacgtc
     3481 gagcagcacg aggtggctgt ggccagatac tgcgacctcc ctagcaaact ggggcacaag
     3541 taagaattct gcagatatcc atcacactgg cggccgctcg agcatgcatc tagagggccc
     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 gggggctccc
     4081 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4141 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4201 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4261 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4321 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     4381 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4441 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4501 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4561 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4621 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4681 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     4741 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4801 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4861 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4921 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4981 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     5041 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     5101 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     5161 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     5221 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     5281 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     5341 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     5401 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     5461 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5521 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     5581 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     5641 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     5701 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     5761 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5821 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     5881 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5941 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     6001 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     6061 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     6121 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     6181 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     6241 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6301 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6361 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6421 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6481 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6541 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6601 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6661 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6721 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6781 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6841 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6901 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6961 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     7021 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     7081 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     7141 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     7201 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     7261 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     7321 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     7381 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     7441 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     7501 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7561 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     7621 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     7681 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7741 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7801 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7861 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7921 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7981 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     8041 cacctgacgt c
//
