LOCUS       sequence-362816-        6924 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Endoplasmic reticulum-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 6924)
  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 6924)
  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-CYP450-uDpTT
FEATURES             Location/Qualifiers
     source          1..6924
                     /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             1025..1696
                     /codon_start=1
                     /product="monomeric photoswitchable Dronpa-Green1 
                     fluorescent protein"
                     /label=Dronpa-Green1
                     /translation="MSVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSMDLKVK
                     EGGPLPFAYDILTTVFCYGNRVFAKYPENIVDYFKQSFPEGYSWERSMNYEDGGICNAT
                     NDITLDGDCYIYEIRFDGVNFPANGPVMQKRTVKWEPSTEKLYVRDGVLKGDVNMALSL
                     EGGGHYRCDFKTTYKAKKVVQLPDYHFVDTHIEIKSHDKDYSNVNLHEHAEAHSELPRQ
                     AK"
     CDS             1721..2416
                     /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(2477..2495)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2478..2495)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2515..2532)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2521..2745
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2791..3219
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2878..2897)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3088..3109
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3228..3248)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3233..3562
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3413..3548
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3475..3494
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3629..4423
                     /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(3683..3702)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4293..4312
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4597..4718
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4634..4653)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4688..4707
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4767..4783)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4767..4783)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4780..4802)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4791..4807
                     /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(4815..4821,4822..4839,4840..4845))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4860..4881
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4998..5015)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5169..5757)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5249..5268)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5928..6719,6720..6788))
                     /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     6551..6570
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6789..6893)
                     /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 gcttggtacc
      901 ccttgcagct tccagttgtc aggaagagaa ggctcccatg gaccctgtgg tggtgctggg
      961 gctctgtctc tcctgtttgc ttctcctttc actctggaaa cagagctatg ggggagggga
     1021 tccgatgagt gtgattaaac cagacatgaa gatcaagctg cgtatggaag gcgctgtaaa
     1081 tggacacccg ttcgcgattg aaggagttgg ccttgggaag cctttcgagg gaaaacagag
     1141 tatggacctt aaagtcaaag aaggcggacc tctgcctttc gcctatgaca tcttgacaac
     1201 tgtgttctgt tacggcaaca gggtattcgc caaataccca gaaaatatag tagactattt
     1261 caagcagtcg tttcctgagg gctactcttg ggaacgaagc atgaattacg aagacggggg
     1321 catttgtaac gcgacaaacg acataaccct ggatggtgac tgttatatct atgaaattcg
     1381 atttgatggt gtgaactttc ctgccaatgg tccagttatg cagaagagga ctgtgaaatg
     1441 ggagccatcc actgagaaat tgtatgtgcg tgatggagtg ctgaagggtg atgttaacat
     1501 ggctctgtcg cttgaaggag gtggccatta ccgatgtgac ttcaaaacta cttataaagc
     1561 taagaaggtt gtccagttgc cagactatca ctttgtggac acacacattg agattaaaag
     1621 ccacgacaaa gattacagta atgttaatct gcatgagcat gccgaagcgc attctgagct
     1681 gccgaggcag gccaagagat ctgacgaggt ggacggtacc atgagcgagc tgattaagga
     1741 gaacatgcac atgaagctgt acatggaggg caccgtgaac aaccaccact tcaagtgcac
     1801 atccgagggc gaaggcaagc cctacgaggg cacccagacc atgagaatca aggtggtcga
     1861 gggcggccct ctccccttcg ccttcgacat cctggctacc agcttcatgt acggcagcag
     1921 aaccttcatc aaccacaccc agggcatccc cgatttcttt aagcagtcct tccctgaggg
     1981 cttcacatgg gagagagtca ccacatacga agacgggggc gtgctgaccg ctacccagga
     2041 caccagcctc caggacggct gcctcatcta caacgtcaag atcagagggg tgaacttccc
     2101 atccaacggc cctgtgatgc agaagaaaac actcggctgg gaggccaaca ccgagatgct
     2161 gtaccccgct gacggcggcc tggaaggcag aaccgacatg gccctgaagc tcgtgggcgg
     2221 gggccacctg atctgcaact tcaagaccac atacagatcc aagaaacccg ctaagaacct
     2281 caagatgccc ggcgtctact atgtggacca cagactggaa agaatcaagg aggccgacaa
     2341 agagacctac gtcgagcagc acgaggtggc tgtggccaga tactgcgacc tccctagcaa
     2401 actggggcac aagtaagaat tctgcagata tccatcacac tggcggccgc tcgagcatgc
     2461 atctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga
     2521 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     2581 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     2641 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     2701 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     2761 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     2821 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     2881 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     2941 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3001 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3061 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3121 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3181 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     3241 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3301 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3361 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3421 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3481 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     3541 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     3601 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     3661 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3721 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3781 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     3841 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3901 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     3961 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4021 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4081 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4141 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4201 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4261 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4321 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4381 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4441 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     4501 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4561 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     4621 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4681 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     4741 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4801 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4861 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4921 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4981 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5041 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5101 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5161 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5221 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5281 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5341 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5401 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5461 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5521 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5581 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5641 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5701 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5761 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5821 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5881 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5941 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6001 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6061 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6121 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6181 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6241 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6301 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6361 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6421 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6481 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6541 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6601 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6661 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6721 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6781 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6841 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6901 ccccgaaaag tgccacctga cgtc
//
