LOCUS       sequence-311951-        8268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Shield-1 inducible transient mammalian expression of Dam-LMNB1 for 
            DamID, containing the wild-type allele of Dam..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8268)
  AUTHORS   Altemose N, Maslan A, Rios-Martinez C, Lai A, White JA, Streets A
  TITLE     muDamID: A Microfluidic Approach for Joint Imaging and Sequencing of
            Protein-DNA Interactions in Single Cells.
  JOURNAL   Cell Syst. 2020 Oct 21;11(4):354-366.e9. doi: 
            10.1016/j.cels.2020.08.015. Epub 2020 Sep 23.
  PUBMED    33099405
REFERENCE   2  (bases 1 to 8268)
  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: 
            "10.1016/j.cels.2020.08.015"; journalName: "Cell Syst"; date: 
            "2020-10-21- 21"; volume: "11"; issue: "4"; pages: "354-366.e9"
COMMENT     Sequence Label: DD-DamWT-LMNB1-IRES2-mCherry
FEATURES             Location/Qualifiers
     source          1..8268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(537..539,540..860)
                     /codon_start=1
                     /product="destabilization domain that can be stabilized by 
                     Shield1 in the ProteoTuner(TM) system"
                     /label=DD
                     /note="L106P mutant of FKBP12"
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKPE"
     CDS             1746..1787
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    1803..1827
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3625..3649)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    join(3681..4255,4256..4258,4259..4267)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3848..3865)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4075..4094
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4268..4978
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4413..4431)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4681..4700
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4996..5117
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5033..5052)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5087..5106
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5124..5579)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5261..5282)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5473..5492
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5606..5710
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5712..6069
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5712..5903
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5735..5755)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5920..6055
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6104..6898
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6104..6898
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6158..6177)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6768..6787
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7086..7105)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7130..7177
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7506..8094
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7995..8014
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcatgg
      541 gagtgcaggt ggaaaccatc tccccaggag acgggcgcac cttccccaag cgcggccaga
      601 cctgtgtggt gcactacacc gggatgcttg aagatggaaa gaaagtcgat tcctcccggg
      661 acagaaacaa gccctttaag tttatgctag gcaagcagga ggtgatccga ggctgggaag
      721 aaggggttgc ccagatgagt gtgggtcaga gagccaaact gactatatct ccagattatg
      781 cctatggtgc cactgggcac ccaggcatca tcccaccaca tgccactctc gtcttcgatg
      841 tggagcttct aaaaccggaa gctagcgcta ccggactcag atcttcacac acagctagcg
      901 ccaccatgaa gaaaaatcgc gcttttttga agtgggcagg gggcaagtat cccctgcttg
      961 atgatattaa acggcatttg cccaagggcg aatgtctggt tgagcctttt gtaggtgccg
     1021 ggtcggtgtt tctcaacacc gacttttctc gttatatcct tgccgatatc aatagcgacc
     1081 tgatcagtct ctataacatt gtgaagatgc gtactgatga gtacgtacag gccgcacgcg
     1141 agctgtttgt tcccgaaaca aattgcgccg aggtttacta tcagttccgc gaagagttca
     1201 acaaaagcca ggatccgttc cgtcgggcgg tactgttttt atatttgaac cgctacggtt
     1261 acaacggcct gtgtcgttac aatctgcgcg gtgagtttaa cgtgccgttc ggccgctaca
     1321 aaaaacccta tttcccggaa gcagagttgt atcacttcgc tgaaaaagcg cagaatgcct
     1381 ttttctattg tgagtcttac gccgatagca tggcgcgcgc agatgatgca tccgtcgtct
     1441 attgcgatcc gccttatgca ccgctgtctg cgaccgccaa ctttacggcg tatcacacaa
     1501 acagttttac gcttgaacaa caagcgcatc tggcggagat cgccgaaggt ctggttgagc
     1561 gccatattcc agtgctgatc tccaatcacg atacgatgtt aacgcgtgag tggtatcagc
     1621 gcgcaaaatt gcatgtcgtc aaagttcgac gcagtataag cagcaacggc ggcacacgta
     1681 aaaaggtgga cgaactgctg gctttgtaca aaccaggagt cgtttcaccc gcgaaaaaaa
     1741 ccggtggtaa gcctatccct aaccctctcc tcggtctcga ttctacgatc gatttcgatc
     1801 aaacaagttt gtacaaaaaa gcaggctccg cggccgcccc cttcaccatg gcgactgcga
     1861 cccccgtgcc gccgcggatg ggcagccgcg ctggcggccc caccacgccg ctgagcccca
     1921 cgcgcctgtc gcggctccag gagaaggagg agctgcgcga gctcaatgac cggctggcgg
     1981 tgtacatcga caaggtgcgc agcctggaga cggagaacag cgcgctgcag ctgcaggtga
     2041 cggagcgcga ggaggtgcgc ggccgtgagc tcaccggcct caaggcgctc tacgagaccg
     2101 agctggccga cgcgcgacgc gcgctcgacg acacggcccg cgagcgcgcc aagctgcaga
     2161 tcgagctggg caagtgcaag gcggaacacg accagctgct cctcaactat gctaagaagg
     2221 aatctgatct taatggcgcc cagatcaagc ttcgagaata tgaagcagca ctgaattcga
     2281 aagatgcagc tcttgctact gcacttggtg acaaaaaaag tttagaggga gatttggagg
     2341 atctgaagga tcagattgcc cagttggaag cctccttagc tgcagccaaa aaacagttag
     2401 cagatgaaac tttacttaaa gtagatttgg agaatcgttg tcagagcctt actgaggact
     2461 tggagtttcg caaaagcatg tatgaagagg agattaacga gaccagaagg aagcatgaaa
     2521 cgcgcttggt agaggtggat tctgggcgtc aaattgagta tgagtacaag ctggcgcaag
     2581 cccttcatga gatgagagag caacatgatg cccaagtgag gctgtataag gaggagctgg
     2641 agcagactta ccatgccaaa cttgagaatg ccagactgtc atcagagatg aatacttcta
     2701 ctgtcaacag tgccagggaa gaactgatgg aaagccgcat gagaattgag agcctttcat
     2761 cccagctttc taatctacag aaagagtcta gagcatgttt ggaaaggatt caagaattag
     2821 aggacttgct tgctaaagaa aaagacaact ctcgtcgcat gctgacagac aaagagagag
     2881 agatggcgga aataagggat caaatgcagc aacagctgaa tgactatgaa cagcttcttg
     2941 atgtaaagtt agccctggac atggaaatca gtgcttacag gaaactctta caaggcgaag
     3001 aagagaggtt gaagctgtct ccaagccctt cttcccgtgt gacagtatcc cgagcatcct
     3061 caagtcgtag tgtacgtaca actagaggaa agcggaagag ggttgatgtg gaagaatcag
     3121 aggcgagtag tagtgttagc atctctcatt ccgcctcagc cactggaaat gtttgcatcg
     3181 aagaaattga tgttgatggg aaatttatcc gcttgaagaa cacttctgaa caggatcaac
     3241 caatgggagg ctgggagatg atcagaaaaa ttggagacac atcagtcagt tataaatata
     3301 cctcaagata tgtgctgaag gcaggccaga ctgttacaat ttgggctgca aacgctggtg
     3361 tcacagccag ccccccaact gacctcatct ggaagaacca gaactcgtgg ggcactggcg
     3421 aagatgtgaa ggttatattg aaaaattctc agggagagga ggttgctcaa agaagtacag
     3481 tctttaaaac aaccatacct gaagaagagg aggaggagga agaagcagct ggagtggttg
     3541 ttgaggaaga acttttccac cagcagggaa ccccaagagc atccaataga agctgtgcaa
     3601 ttatgtaaaa gggtgggcgc gccgacccag ctttcttgta caaagtggtt cggtcgacgg
     3661 taccgcgggc ccgggatccg cccctctccc tccccccccc ctaacgttac tggccgaagc
     3721 cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct
     3781 tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt
     3841 ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct
     3901 ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc
     3961 ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag
     4021 gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc
     4081 tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga
     4141 tctgatctgg ggcctcggta cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt
     4201 ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc
     4261 cacaaccatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg
     4321 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
     4381 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc
     4441 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt
     4501 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg
     4561 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
     4621 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
     4681 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga
     4741 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta
     4801 cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
     4861 caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     4921 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaatg
     4981 acaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5041 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5101 ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     5161 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     5221 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     5281 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     5341 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     5401 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     5461 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     5521 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     5581 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     5641 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     5701 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     5761 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     5821 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     5881 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     5941 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     6001 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     6061 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     6121 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     6181 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     6241 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     6301 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     6361 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     6421 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     6481 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     6541 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     6601 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     6661 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     6721 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     6781 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     6841 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     6901 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     6961 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     7021 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     7081 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     7141 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     7201 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     7261 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     7321 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     7381 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     7441 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     7501 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     7561 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     7621 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     7681 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     7741 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     7801 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     7861 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     7921 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     7981 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     8041 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     8101 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     8161 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata
     8221 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccg
//
