LOCUS       sequence-308617-        6415 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Shield-1 inducible transient mammalian expression of Dam alone for 
            DamID controls, containing the wild-type allele 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-DamWT-IRES2-mCherry
FEATURES             Location/Qualifiers
     source          1..6415
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(597..599,600..920)
                     /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             1806..1847
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    join(1888..2462,2463..2465,2466..2474)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2055..2072)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2282..2301
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2475..3185
                     /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(2620..2638)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2888..2907
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3203..3324
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3240..3259)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3294..3313
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3331..3786)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3468..3489)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3680..3699
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3813..3917
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3919..4276
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3919..4110
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3942..3962)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4127..4262
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4311..5105
                     /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             4311..5105
                     /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(4365..4384)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4975..4994
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5293..5312)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5337..5384
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5713..6301
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6202..6221
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcatgg
      601 gagtgcaggt ggaaaccatc tccccaggag acgggcgcac cttccccaag cgcggccaga
      661 cctgtgtggt gcactacacc gggatgcttg aagatggaaa gaaagtcgat tcctcccggg
      721 acagaaacaa gccctttaag tttatgctag gcaagcagga ggtgatccga ggctgggaag
      781 aaggggttgc ccagatgagt gtgggtcaga gagccaaact gactatatct ccagattatg
      841 cctatggtgc cactgggcac ccaggcatca tcccaccaca tgccactctc gtcttcgatg
      901 tggagcttct aaaaccggaa gctagcgcta ccggactcag atcttcacac acagctagcg
      961 ccaccatgaa gaaaaatcgc gcttttttga agtgggcagg gggcaagtat cccctgcttg
     1021 atgatattaa acggcatttg cccaagggcg aatgtctggt tgagcctttt gtaggtgccg
     1081 ggtcggtgtt tctcaacacc gacttttctc gttatatcct tgccgatatc aatagcgacc
     1141 tgatcagtct ctataacatt gtgaagatgc gtactgatga gtacgtacag gccgcacgcg
     1201 agctgtttgt tcccgaaaca aattgcgccg aggtttacta tcagttccgc gaagagttca
     1261 acaaaagcca ggatccgttc cgtcgggcgg tactgttttt atatttgaac cgctacggtt
     1321 acaacggcct gtgtcgttac aatctgcgcg gtgagtttaa cgtgccgttc ggccgctaca
     1381 aaaaacccta tttcccggaa gcagagttgt atcacttcgc tgaaaaagcg cagaatgcct
     1441 ttttctattg tgagtcttac gccgatagca tggcgcgcgc agatgatgca tccgtcgtct
     1501 attgcgatcc gccttatgca ccgctgtctg cgaccgccaa ctttacggcg tatcacacaa
     1561 acagttttac gcttgaacaa caagcgcatc tggcggagat cgccgaaggt ctggttgagc
     1621 gccatattcc agtgctgatc tccaatcacg atacgatgtt aacgcgtgag tggtatcagc
     1681 gcgcaaaatt gcatgtcgtc aaagttcgac gcagtataag cagcaacggc ggcacacgta
     1741 aaaaggtgga cgaactgctg gctttgtaca aaccaggagt cgtttcaccc gcgaaaaaaa
     1801 ccggtggtaa gcctatccct aaccctctcc tcggtctcga ttctacgatc gggtaatgag
     1861 tcgacggtac cgcgggcccg ggatccgccc ctctccctcc ccccccccta acgttactgg
     1921 ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt
     1981 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc
     2041 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc
     2101 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg
     2161 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc
     2221 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa
     2281 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg
     2341 tatgggatct gatctggggc ctcggtacac atgctttaca tgtgtttagt cgaggttaaa
     2401 aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata
     2461 atatggccac aaccatggtg agcaagggcg aggaggataa catggccatc atcaaggagt
     2521 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
     2581 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     2641 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg
     2701 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct
     2761 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
     2821 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct
     2881 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt
     2941 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
     3001 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
     3061 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg
     3121 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     3181 agtaatgaca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     3241 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3301 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
     3361 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     3421 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     3481 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     3541 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     3601 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     3661 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     3721 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     3781 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3841 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3901 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     3961 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4021 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4081 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4141 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4201 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4261 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     4321 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4381 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4441 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     4501 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4561 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4621 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4681 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4741 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4801 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     4861 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4921 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4981 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     5041 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     5101 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     5161 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     5221 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     5281 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     5341 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     5401 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     5461 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     5521 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     5581 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5641 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5701 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5761 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5821 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     5881 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5941 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6001 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6061 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6121 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6181 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6241 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6301 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6361 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
