LOCUS       sequence-381987-        6092 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MPP8-delta2-441_10xHis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6092)
  AUTHORS   Leisegang MS, Bains JK, Seredinski S, Oo JA, Krause NM, Kuo CC, 
            Gunther S, Senturk Cetin N, Warwick T, Cao C, Boos F, Izquierdo 
            Ponce J, Haydar S, Bednarz R, Valasarajan C, Fuhrmann DC, Preussner 
            J, Looso M, Pullamsetti SS, Schulz MH, Jonker HRA, Richter C, 
            Rezende F, Gilsbach R, Pfluger-Muller B, Wittig I, Grummt I, 
            Ribarska T, Costa IG, Schwalbe H, Brandes RP
  TITLE     HIF1alpha-AS1 is a DNA:DNA:RNA triplex-forming lncRNA interacting 
            with the HUSH complex.
  JOURNAL   Nat Commun. 2022 Nov 2;13(1):6563. doi: 10.1038/s41467-022-34252-2.
  PUBMED    36323673
REFERENCE   2  (bases 1 to 6092)
  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: "Nat 
            Commun."; date: "2022-11-2"; volume: "13(1):6563. doi"; pages: " 
            10.1038/s41467-022-34252-2"
COMMENT     Sequence Label: pCMV6-MPP8-delta2-441-10xHis
FEATURES             Location/Qualifiers
     source          1..6092
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..173
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     165..182
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     166..182
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        343..722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        723..926
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     877..897
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     952..971
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        952..970
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2303..2332
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(2349..2365)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2349..2365)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2376..2998
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2575..2594)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(3147..3735)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3227..3246)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4064..4111
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4136..4155
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4343..5137)
                     /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             complement(4343..5137)
                     /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(4454..4473)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5064..5083
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5172..5529)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5186..5321
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5338..5529
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5486..5506
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5531..5635)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5662..6090
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5749..5768)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5959..5980
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccggga attcgtcgac tggatccagt accgaggaga tctgcgccgc
     1021 gatcgccatg aaggaaatca gaaatgcatt tgatttattt aaattaactc cagaagaaaa
     1081 aaatgatgtt tctgagaata atcggaaaag ggaagaaata ccactggatt ttaaaaccat
     1141 agacgatcac aaaaccaagg aaaacaaaca gtcacttaaa gaaaggagaa acaccagaga
     1201 cgaaacggat acttgggcat acattgctgc agaaggtgat caggaggttt tagacagcgt
     1261 gtgccaagca gatgagaatt cagatggcag gcagcagatt ctgagtttgg gcatggacct
     1321 gcagttggaa tggatgaagt tggaagattt ccaaaagcac cttgatggga aagatgagaa
     1381 ttttgctgca acagatgcaa ttccaagtaa tgtgttaagg gatgctgtga aaaatgggga
     1441 ttatattact gtaaaagttg cacttaattc aaatgaagaa tataacctgg accaagagga
     1501 ttccagtgga atgacactgg tgatgcttgc cgccgccgga gggcaggacg acctcctgcg
     1561 actcctcatc acaaaaggcg cgaaagtgaa cggtcggcag aagaacggga ccaccgccct
     1621 cattcatgct gcagagaaga actttttaac aacagtggct attcttttgg aagcaggagc
     1681 ttttgtaaat gtccagcaaa gcaatggtga gactgcactg atgaaggcct gtaaaagagg
     1741 aaattcagac atcgtacgac tcgtaattga atgtggagct gactgcaata ttttgtcaaa
     1801 gcaccagaat agtgccctgc actttgcgaa gcagtctaac aatgtgcttg tgtacgactt
     1861 gctgaagaac catttagaga cactttcaag agtagcagaa gagacaataa aggattactt
     1921 tgaagctcgc cttgctctgc tagaaccagt ttttccaatc gcatgtcatc gactctgtga
     1981 gggtccagat ttttcaacag atttcaatta caaaccccca cagaacatac cagaaggctc
     2041 tggcatcctg ctgtttatct tccatgcaaa ctttttgggt aaagaagtta ttgctcggct
     2101 ctgtggaccg tgtagtgtac aagctgtagt tctgaatgat aaatttcagc ttcctgtttt
     2161 tctggacagt cattttgttt actcattcag ccctgttgca ggtcccaata aactcttcat
     2221 aaggttgaca gaagcaccct ctgccaaggt taagttgcta ataggtgcat acagagtgca
     2281 gctgcagggg ggtggaggct ctcaccatca ccaccatcat caccaccatc actaaacggc
     2341 cggccgcggt catagctgtt tcctgaacag atcccgggtg gcatccctgt gacccctccc
     2401 cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa
     2461 aattaagttg catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg
     2521 gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt
     2581 gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg
     2641 ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca
     2701 ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg
     2761 tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac
     2821 aggcgtgaac cactgctccc ttccctgtcc ttctgatttt aaaataacta taccagcagg
     2881 aggacgtcca gacacagcat aggctacctg gccatgccca accggtggga catttgagtt
     2941 gcttgcttgg cactgtcctc tcatgcgttg ggtccactca gtagatgcct gttgaattgg
     3001 gtacgcggcc agcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3061 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3121 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3181 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3241 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3301 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3361 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3421 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3481 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3541 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3601 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3661 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3721 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3781 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3841 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta acctgaggct
     3901 atggcagggc ctgccgcccc gacgttggct gcgagccctg ggccttcacc cgaacttggg
     3961 gggtggggtg gggaaaagga agaaacgcgg gcgtattggc cccaatgggg tctcggtggg
     4021 gtatcgacag agtgccagcc ctgggaccga accccgcgtt tatgaacaaa cgacccaaca
     4081 ccgtgcgttt tattctgtct ttttattgcc gtcatagcgc gggttccttc cggtattgtc
     4141 tccttccgtg tttcagttag cctcccccta gggtgggcga agaactccag catgagatcc
     4201 ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca
     4261 tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc
     4321 atttcgaacc ccagagtccc gctcagaaga actcgtcaag aaggcgatag aaggcgatgc
     4381 gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc
     4441 caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcga tccgccacac
     4501 ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca
     4561 agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgctc gccttgagcc
     4621 tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga
     4681 caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga
     4741 atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata
     4801 ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata
     4861 gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg
     4921 tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca
     4981 ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat
     5041 cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg
     5101 ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct catcctgtct
     5161 cttgatcgat ctttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat
     5221 agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag tcagccatgg
     5281 ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg
     5341 actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
     5401 ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct
     5461 gctggggagc ctggggactt tccacaccct aactgacaca cattccacag ctggttcttt
     5521 ccgcctcagg actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5581 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5641 ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     5701 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     5761 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     5821 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     5881 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     5941 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     6001 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     6061 agctgattta acaaaaattt aacgcgaatt tt
//
