LOCUS       sequence-311536-        6013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses fusion of ECFP-LacI and HOXD13+7A IDR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6013)
  AUTHORS   Basu S, Mackowiak SD, Niskanen H, Knezevic D, Asimi V, Grosswendt S,
            Geertsema H, Ali S, Jerkovic I, Ewers H, Mundlos S, Meissner A, 
            Ibrahim DM, Hnisz D
  TITLE     Unblending of Transcriptional Condensates in Human Repeat Expansion 
            Disease.
  JOURNAL   Cell. 2020 May 28;181(5):1062-1079.e30. doi: 
            10.1016/j.cell.2020.04.018. Epub 2020 May 7.
  PUBMED    32386547
REFERENCE   2  (bases 1 to 6013)
  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.cell.2020.04.018"; journalName: "Cell"; date: 
            "2020-05-28- 28"; volume: "181"; issue: "5"; pages: "1062-1079.e30"
COMMENT     Sequence Label: pCFP-LacI-HOXD13(+7A)-IDR
FEATURES             Location/Qualifiers
     source          1..6013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..330
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      181..316
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      349..358
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(355..357,358..360,361..1071)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(400..421)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(661..680)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1008..1029
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1109..1128)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2170..2190
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2783..2904
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2820..2839)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2874..2893
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2911..3366)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3048..3069)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3260..3279
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3393..3497
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3499..3856
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3499..3690
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     rep_origin      3707..3842
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3891..4685
                     /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             3891..4685
                     /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(3945..3964)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4555..4574
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4873..4892)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4917..4964
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5293..5881
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5782..5801
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
       61 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
      121 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
      181 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
      241 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
      301 ggcttttttg gaggcctagg cttttgcaaa aagctagcgc taccggtcgc caccatggtg
      361 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
      421 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
      481 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
      541 accaccctga cctggggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
      601 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
      661 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
      721 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
      781 gagtacaact acatcagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc
      841 aaggccaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
      901 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
      961 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1021 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtccggactc
     1081 agatccatgg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat
     1141 cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa
     1201 gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg
     1261 ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg
     1321 caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg
     1381 atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa
     1441 cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa
     1501 gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac
     1561 agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg
     1621 ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt
     1681 ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa
     1741 ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc
     1801 gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt
     1861 accgagtccg ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa
     1921 gacagctcat gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg
     1981 caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag
     2041 ctgttgcccg tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc
     2101 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgaagcagc
     2161 ctgaggcctc ctaagaagaa gaggaaggtt ggtgcaccgg gttctgcggg ttctgccgca
     2221 ggtggatccg cgatcgcttc ccgagccggt agctgggaca tggacggcct gcgagcggac
     2281 ggaggcggag ctggaggagc tcccgcatca agtagtagct caagtgtggc cgctgccgcg
     2341 gctagcggac aatgtagggg gtttttgtca gcgcctgtct ttgcgggcac acattccggg
     2401 agggccgctg ctgcggctgc tgctgctgct gcagccgcgg cggcagctgc agctgcggcc
     2461 gctgccgcta gtggatttgc ctatcctggg acgagtgagc gcactggttc atcatcctca
     2521 tcttcatcca gtgcggtagt cgccgctcgg ccagaagcac cacctgcaaa agagtgtccg
     2581 gccccgacgc cagctgcagc agcagcggca ccgccttcag ctcctgcgtg acgtacgtct
     2641 agagggcccg aattcgagcg ccgtcgacaa gcttgcggcc gcaatcagcc ataccacatt
     2701 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     2761 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     2821 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     2881 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
     2941 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     3001 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     3061 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     3121 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     3181 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     3241 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     3301 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     3361 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3421 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3481 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     3541 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3601 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3661 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3721 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3781 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3841 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3901 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3961 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4021 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     4081 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4141 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4201 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4261 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4321 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4381 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     4441 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4501 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4561 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4621 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4681 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4741 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4801 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     4861 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     4921 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     4981 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     5041 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     5101 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     5161 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5221 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5281 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5341 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5401 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     5461 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5521 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5581 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5641 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5701 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5761 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5821 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5881 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5941 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt
     6001 attactgtgg aat
//
