LOCUS       sequence-204230-        7458 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human ASXL1 (amino acids 1-115) fused to the N-terminal of
            firefly luciferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7458)
  AUTHORS   Kweon SM, Chen Y, Moon E, Kvederaviciute K, Klimasauskas S, Feldman 
            DE
  TITLE     An Adversarial DNA N(6)-Methyladenine-Sensor Network Preserves 
            Polycomb Silencing.
  JOURNAL   Mol Cell. 2019 Apr 5. pii: S1097-2765(19)30222-9. doi: 
            10.1016/j.molcel.2019.03.018.
  PUBMED    30982744
REFERENCE   2  (bases 1 to 7458)
  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"; journalName: "Mol Cell. 
            2019 Apr 5. pii: S1097-2765(19)30222-9. doi: 
            10.1016/j.molcel.2019.03.018."
COMMENT     Sequence Label: hASXL(1-115)-luciferase
FEATURES             Location/Qualifiers
     source          1..7458
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1042..2694
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(1108..1126)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     promoter        complement(2777..2795)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2778..2795)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2815..2832)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2821..3045
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3091..3519
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3178..3197)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3388..3409
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3528..3548)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3533..3862
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3713..3848
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3775..3794
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3929..4723
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3983..4002)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4593..4612
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4897..5018
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4934..4953)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4988..5007
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5067..5083)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5067..5083)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5080..5102)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5091..5107
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5115..5121,5122..5139,5140..5145))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5160..5181
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5298..5315)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5469..6057)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5549..5568)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6228..7019,7020..7088))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6851..6870
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7089..7193)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7268..7287)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 gccatggcaa aggacaaaca gaagaagaag aaggagcgca cgtgggccga ggccgcgcgc
      721 ctggtactag aaaactactc ggatgctcca atgacaccaa aacagattct gcaggtcata
      781 gaggcagaag gactaaagga aatgagaagt gggacttccc ctctcgcatg cctcaatgct
      841 atgctacatt ccaattcaag aggaggagag gggttgtttt ataaactgcc tggccgaatc
      901 agccttttca cgctcaagaa ggatgccctg cagtggtctc gccatccagc tacagtggag
      961 ggagaggagc cagaggacac ggctgatgtg gagagctgtg ggtctaatga agccgaagct
     1021 tggtaccgag ctcggatcca aatggaagac gccaaaaaca taaagaaagg cccggcgcca
     1081 ttctatcctc tagaggatgg aaccgctgga gagcaactgc ataaggctat gaagagatac
     1141 gccctggttc ctggaacaat tgcttttaca gatgcacata tcgaggtgaa catcacgtac
     1201 gcggaatact tcgaaatgtc cgttcggttg gcagaagcta tgaaacgata tgggctgaat
     1261 acaaatcaca gaatcgtcgt atgcagtgaa aactctcttc aattctttat gccggtgttg
     1321 ggcgcgttat ttatcggagt tgcagttgcg cccgcgaacg acatttataa tgaacgtgaa
     1381 ttgctcaaca gtatgaacat ttcgcagcct accgtagtgt ttgtttccaa aaaggggttg
     1441 caaaaaattt tgaacgtgca aaaaaaatta ccaataatcc agaaaattat tatcatggat
     1501 tctaaaacgg attaccaggg atttcagtcg atgtacacgt tcgtcacatc tcatctacct
     1561 cccggtttta atgaatacga ttttgtacca gagtcctttg atcgtgacaa aacaattgca
     1621 ctgataatga attcctctgg atctactggg ttacctaagg gtgtggccct tccgcataga
     1681 actgcctgcg tcagattctc gcatgccaga gatcctattt ttggcaatca aatcattccg
     1741 gatactgcga ttttaagtgt tgttccattc catcacggtt ttggaatgtt tactacactc
     1801 ggatatttga tatgtggatt tcgagtcgtc ttaatgtata gatttgaaga agagctgttt
     1861 ttacgatccc ttcaggatta caaaattcaa agtgcgttgc tagtaccaac cctattttca
     1921 ttcttcgcca aaagcactct gattgacaaa tacgatttat ctaatttaca cgaaattgct
     1981 tctgggggcg cacctctttc gaaagaagtc ggggaagcgg ttgcaaaacg cttccatctt
     2041 ccagggatac gacaaggata tgggctcact gagactacat cagctattct gattacaccc
     2101 gagggggatg ataaaccggg cgcggtcggt aaagttgttc cattttttga agcgaaggtt
     2161 gtggatctgg ataccgggaa aacgctgggc gttaatcaga gaggcgaatt atgtgtcaga
     2221 ggacctatga ttatgtccgg ttatgtaaac aatccggaag cgaccaacgc cttgattgac
     2281 aaggatggat ggctacattc tggagacata gcttactggg acgaagacga acacttcttc
     2341 atagttgacc gcttgaagtc tttaattaaa tacaaaggat atcaggtggc ccccgctgaa
     2401 ttggaatcga tattgttaca acaccccaac atcttcgacg cgggcgtggc aggtcttccc
     2461 gacgatgacg ccggtgaact tcccgccgcc gttgttgttt tggagcacgg aaagacgatg
     2521 acggaaaaag agatcgtgga ttacgtcgcc agtcaagtaa caaccgcgaa aaagttgcgc
     2581 ggaggagttg tgtttgtgga cgaagtaccg aaaggtctta ccggaaaact cgacgcaaga
     2641 aaaatcagag agatcctcat aaaggccaag aagggcggaa agtccaaatt gtaaaatgta
     2701 actgtattca gcgatgacga aattcttagc tattgtaatc ctccgaggcc tcgagcatgc
     2761 atctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga
     2821 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     2881 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     2941 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     3001 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     3061 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     3121 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3181 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3241 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3301 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3361 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3421 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3481 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     3541 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3601 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3661 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3721 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3781 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     3841 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     3901 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     3961 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4021 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4081 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     4141 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4201 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4261 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4321 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4381 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4441 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4501 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4561 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4621 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4681 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4741 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     4801 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4861 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     4921 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4981 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     5041 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     5101 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     5161 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5221 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5281 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5341 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5401 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5461 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5521 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5581 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5641 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5701 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5761 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5821 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5881 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5941 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6001 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6061 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6121 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6181 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6241 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6301 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6361 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6421 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6481 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6541 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6601 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6661 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6721 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6781 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6841 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6901 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6961 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7021 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7081 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7141 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7201 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
     7261 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     7321 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     7381 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
     7441 gggccagata tacgcgtt
//
