LOCUS       sequence-311534-        5992 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses fusion of ECFP-LacI and HOXD13wt IDR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5992)
  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 5992)
  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-HOXD13wt-IDR
FEATURES             Location/Qualifiers
     source          1..5992
                     /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    2762..2883
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2799..2818)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2853..2872
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2890..3345)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3027..3048)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3239..3258
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3372..3476
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3478..3835
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3478..3669
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     rep_origin      3686..3821
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3870..4664
                     /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             3870..4664
                     /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(3924..3943)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4534..4553
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4852..4871)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4896..4943
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5272..5860
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5761..5780
                     /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 cggccgctag tggatttgcc
     2461 tatcctggga cgagtgagcg cactggttca tcatcctcat cttcatccag tgcggtagtc
     2521 gccgctcggc cagaagcacc acctgcaaaa gagtgtccgg ccccgacgcc agctgcagca
     2581 gcagcggcac cgccttcagc tcctgcgtga cgtacgtcta gagggcccga attcgagcgc
     2641 cgtcgacaag cttgcggccg caatcagcca taccacattt gtagaggttt tacttgcttt
     2701 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     2761 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     2821 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     2881 ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     2941 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     3001 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     3061 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     3121 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     3181 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     3241 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     3301 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     3361 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3421 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     3481 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3541 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3601 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3661 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3721 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3781 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     3841 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     3901 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3961 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4021 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     4081 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4141 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4201 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4261 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4321 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4381 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4441 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4501 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4561 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4621 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4681 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4741 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4801 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     4861 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     4921 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     4981 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     5041 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     5101 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     5161 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5221 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5281 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5341 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5401 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5461 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5521 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5581 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5641 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5701 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5761 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5821 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5881 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     5941 tgattctgtg gataaccgta ttaccgccat gcattagtta ttactgtgga at
//
