LOCUS       sequence-311948-        6415 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Shield-1 inducible transient mammalian expression of Dam alone for 
            DamID controls, containing the V133A mutant of Dam..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6415)
  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 6415)
  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-DamV133A-IRES2-mCherry
FEATURES             Location/Qualifiers
     source          1..6415
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(78..652,653..655,656..664)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(245..262)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     472..491
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             665..1375
                     /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(810..828)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1078..1097
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1393..1514
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1430..1449)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1484..1503
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1521..1976)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1658..1679)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1870..1889
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2003..2107
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2109..2466
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2109..2300
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2132..2152)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2317..2452
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2501..3295
                     /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             2501..3295
                     /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(2555..2574)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3165..3184
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3483..3502)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3527..3574
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3903..4491
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4392..4411
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4666..4969
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4970..5173
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5124..5144
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(5202..5204,5205..5525)
                     /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             join(6411..6415,1..37)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
ORIGIN
        1 gcctatccct aaccctctcc tcggtctcga ttctacgatc gggtaatgag tcgacggtac
       61 cgcgggcccg ggatccgccc ctctccctcc ccccccccta acgttactgg ccgaagccgc
      121 ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt
      181 ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt
      241 tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg
      301 gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca
      361 cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg
      421 gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc
      481 tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct
      541 gatctggggc ctcggtacac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta
      601 ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac
      661 aaccatggtg agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt
      721 caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga
      781 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct
      841 gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa
      901 gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga
      961 gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca
     1021 ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc
     1081 cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga
     1141 cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga
     1201 cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa
     1261 cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta
     1321 cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca agtaatgaca
     1381 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     1441 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     1501 atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     1561 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     1621 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     1681 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     1741 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     1801 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     1861 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     1921 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     1981 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     2041 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     2101 gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     2161 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2221 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2281 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     2341 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     2401 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     2461 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     2521 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     2581 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     2641 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
     2701 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     2761 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     2821 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     2881 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     2941 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     3001 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
     3061 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     3121 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     3181 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     3241 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     3301 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     3361 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     3421 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
     3481 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     3541 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     3601 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     3661 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     3721 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
     3781 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     3841 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3901 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3961 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4021 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4081 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4141 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4201 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4261 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4321 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4381 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4441 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4501 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     4561 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta
     4621 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     4681 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     4741 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     4801 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     4861 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     4921 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     4981 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     5041 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     5101 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     5161 tataagcaga gctggtttag tgaaccgtca gatccgctag catgggagtg caggtggaaa
     5221 ccatctcccc aggagacggg cgcaccttcc ccaagcgcgg ccagacctgt gtggtgcact
     5281 acaccgggat gcttgaagat ggaaagaaag tcgattcctc ccgggacaga aacaagccct
     5341 ttaagtttat gctaggcaag caggaggtga tccgaggctg ggaagaaggg gttgcccaga
     5401 tgagtgtggg tcagagagcc aaactgacta tatctccaga ttatgcctat ggtgccactg
     5461 ggcacccagg catcatccca ccacatgcca ctctcgtctt cgatgtggag cttctaaaac
     5521 cggaagctag cgctaccgga ctcagatctt cacacacagc tagcgccacc atgaagaaaa
     5581 atcgcgcttt tttgaagtgg gcagggggca agtatcccct gcttgatgat attaaacggc
     5641 atttgcccaa gggcgaatgt ctggttgagc cttttgtagg tgccgggtcg gtgtttctca
     5701 acaccgactt ttctcgttat atccttgccg atatcaatag cgacctgatc agtctctata
     5761 acattgtgaa gatgcgtact gatgagtacg tacaggccgc acgcgagctg tttgttcccg
     5821 aaacaaattg cgccgaggtt tactatcagt tccgcgaaga gttcaacaaa agccaggatc
     5881 cgttccgtcg ggcggtactg tttttatatt tgaaccgcta cggttacaac ggcctgtgtc
     5941 gttacaatct gcgcggtgag tttaacgcgc cgttcggccg ctacaaaaaa ccctatttcc
     6001 cggaagcaga gttgtatcac ttcgctgaaa aagcgcagaa tgcctttttc tattgtgagt
     6061 cttacgccga tagcatggcg cgcgcagatg atgcatccgt cgtctattgc gatccgcctt
     6121 atgcaccgct gtctgcgacc gccaacttta cggcgtatca cacaaacagt tttacgcttg
     6181 aacaacaagc gcatctggcg gagatcgccg aaggtctggt tgagcgccat attccagtgc
     6241 tgatctccaa tcacgatacg atgttaacgc gtgagtggta tcagcgcgca aaattgcatg
     6301 tcgtcaaagt tcgacgcagt ataagcagca acggcggcac acgtaaaaag gtggacgaac
     6361 tgctggcttt gtacaaacca ggagtcgttt cacccgcgaa aaaaaccggt ggtaa
//
