LOCUS       sequence-363396-        7190 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Histone H2B-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 7190)
  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 7190)
  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-H2B-uDpTT
FEATURES             Location/Qualifiers
     source          1..7190
                     /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             907..1284
                     /codon_start=1
                     /gene="HIST1H2BB"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPSKSAPAPKKGSKKAITKAQKKDGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAVTKYTSSK"
     CDS             1291..1962
                     /codon_start=1
                     /product="monomeric photoswitchable Dronpa-Green1 
                     fluorescent protein"
                     /label=Dronpa-Green1
                     /translation="MSVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSMDLKVK
                     EGGPLPFAYDILTTVFCYGNRVFAKYPENIVDYFKQSFPEGYSWERSMNYEDGGICNAT
                     NDITLDGDCYIYEIRFDGVNFPANGPVMQKRTVKWEPSTEKLYVRDGVLKGDVNMALSL
                     EGGGHYRCDFKTTYKAKKVVQLPDYHFVDTHIEIKSHDKDYSNVNLHEHAEAHSELPRQ
                     AK"
     CDS             1987..2682
                     /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(2743..2761)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2744..2761)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2781..2798)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2787..3011
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3057..3485
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3144..3163)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3354..3375
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3494..3514)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3499..3828
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3679..3814
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3741..3760
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3895..4689
                     /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(3949..3968)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4559..4578
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4863..4984
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4900..4919)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4954..4973
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5033..5049)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5033..5049)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5046..5068)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5057..5073
                     /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(5081..5087,5088..5105,5106..5111))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5126..5147
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5264..5281)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5435..6023)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5515..5534)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6194..6985,6986..7054))
                     /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     6817..6836
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7055..7159)
                     /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 gccaccatgc ctgaaccctc taagtctgct ccagccccta aaaagggttc taagaaggct
      961 atcactaagg cgcagaagaa ggatggtaag aagcgtaagc gcagccgcaa ggagagctat
     1021 tctatctatg tgtacaaggt tctgaagcag gtccaccccg acaccggcat ctcatccaag
     1081 gccatgggga tcatgaactc cttcgtcaac gacatcttcg agcgcatcgc gggcgaggct
     1141 tctcgcctgg ctcactacaa taagcgctcg accatcacct ccagggagat tcagacggct
     1201 gtgcgcctgc tgctgcctgg ggagctggct aagcatgctg tgtccgaggg cactaaggca
     1261 gttaccaagt acactagctc taaggatccg atgagtgtga ttaaaccaga catgaagatc
     1321 aagctgcgta tggaaggcgc tgtaaatgga cacccgttcg cgattgaagg agttggcctt
     1381 gggaagcctt tcgagggaaa acagagtatg gaccttaaag tcaaagaagg cggacctctg
     1441 cctttcgcct atgacatctt gacaactgtg ttctgttacg gcaacagggt attcgccaaa
     1501 tacccagaaa atatagtaga ctatttcaag cagtcgtttc ctgagggcta ctcttgggaa
     1561 cgaagcatga attacgaaga cgggggcatt tgtaacgcga caaacgacat aaccctggat
     1621 ggtgactgtt atatctatga aattcgattt gatggtgtga actttcctgc caatggtcca
     1681 gttatgcaga agaggactgt gaaatgggag ccatccactg agaaattgta tgtgcgtgat
     1741 ggagtgctga agggtgatgt taacatggct ctgtcgcttg aaggaggtgg ccattaccga
     1801 tgtgacttca aaactactta taaagctaag aaggttgtcc agttgccaga ctatcacttt
     1861 gtggacacac acattgagat taaaagccac gacaaagatt acagtaatgt taatctgcat
     1921 gagcatgccg aagcgcattc tgagctgccg aggcaggcca agagatctga cgaggtggac
     1981 ggtaccatga gcgagctgat taaggagaac atgcacatga agctgtacat ggagggcacc
     2041 gtgaacaacc accacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc
     2101 cagaccatga gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg
     2161 gctaccagct tcatgtacgg cagcagaacc ttcatcaacc acacccaggg catccccgat
     2221 ttctttaagc agtccttccc tgagggcttc acatgggaga gagtcaccac atacgaagac
     2281 gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct catctacaac
     2341 gtcaagatca gaggggtgaa cttcccatcc aacggccctg tgatgcagaa gaaaacactc
     2401 ggctgggagg ccaacaccga gatgctgtac cccgctgacg gcggcctgga aggcagaacc
     2461 gacatggccc tgaagctcgt gggcgggggc cacctgatct gcaacttcaa gaccacatac
     2521 agatccaaga aacccgctaa gaacctcaag atgcccggcg tctactatgt ggaccacaga
     2581 ctggaaagaa tcaaggaggc cgacaaagag acctacgtcg agcagcacga ggtggctgtg
     2641 gccagatact gcgacctccc tagcaaactg gggcacaagt aagaattctg cagatatcca
     2701 tcacactggc ggccgctcga gcatgcatct agagggccct attctatagt gtcacctaaa
     2761 tgctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     2821 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     2881 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     2941 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     3001 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     3061 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3121 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3181 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3241 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3301 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3361 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3421 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3481 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3541 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3601 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3661 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3721 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     3781 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     3841 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3901 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3961 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4021 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4081 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4141 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4201 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4261 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4321 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4381 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4441 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4501 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4561 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4621 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4681 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4741 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4801 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4861 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4921 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4981 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     5041 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5101 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5161 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5221 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5281 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5341 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5401 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5461 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5521 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5581 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5641 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5701 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5761 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5821 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5881 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5941 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6001 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6061 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6121 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6181 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6241 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6301 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6361 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6421 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6481 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6541 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6601 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6661 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6721 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6781 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6841 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6901 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6961 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7021 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7081 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7141 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
