LOCUS       sequence-382022-        7412 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MPP8-W80A_10xHis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7412)
  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 7412)
  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-W80A-10xHis
FEATURES             Location/Qualifiers
     source          1..7412
                     /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             3623..3652
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(3669..3685)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3669..3685)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3696..4318
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3895..3914)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(4467..5055)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4547..4566)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5384..5431
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5456..5475
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5663..6457)
                     /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(5663..6457)
                     /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(5774..5793)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6384..6403
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(6492..6849)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6506..6641
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6658..6849
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6806..6826
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(6851..6955)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6982..7410
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7069..7088)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7279..7300
                     /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 gagcaggttg cggagggagc aagggtgacc gcagtccctg tgtcagctgc
     1081 cgacagcact gaggagttgg ccgaagtcga agaaggagtt ggagtagtgg gcgaagataa
     1141 tgacgcagcc gcgagaggag cggaggcctt tggcgacagt gaggaggacg gagaggatgt
     1201 gttcgaggtg gagaagatcc tggacatgaa gaccgagggg ggtaaagttc tttacaaagt
     1261 tcgcgcgaaa ggctatacat cggatgatga tacctgggag cccgagattc acctggagga
     1321 ctgtaaagaa gtgcttcttg aatttaggaa gaaaattgca gagaacaaag ccaaagcagt
     1381 caggaaggat attcagagac tatccttaaa taacgacata tttgaggcga actctgatag
     1441 cgatcagcaa agtgagacaa aagaagatac ttccccaaag aagaaaaaga aaaaattgag
     1501 gcagagagaa gagaaaagcc cagatgatct gaaaaagaaa aaagcaaagg ccgggaagct
     1561 aaaagacaag tccaaaccag acctggagag ctccttggaa agtttagttt ttgatttaag
     1621 gacaaagaaa agaatttctg aagccaaaga agaactaaag gagtccaaaa agcccaaaaa
     1681 agatgaagta aaagaaacaa aagaattaaa gaaagttaaa aagggtgaaa taagagattt
     1741 aaagacgaaa acaagagaag atcccaaaga aaatagaaaa acaaaaaaag aaaaatttgt
     1801 cgaatcccag gtggaatctg aatcaagtgt acttaatgat tctccctttc cagaggatga
     1861 cagtgaaggg ctacattccg acagcagaga agagaaacaa aacactaaaa gtgcaagaga
     1921 gagagcaggg caggacatgg ggctggagca tggctttgag aagcccctag acagtgccat
     1981 gagtgctgag gaggataccg atgtcagagg caggaggaaa aagaagaccc cgagaaaggc
     2041 tgaggacact agagagaaca ggaagctaga gaacaagaac gctttcttag agaagaaaac
     2101 tgtgcctaaa aagcagagga atcaagacag aagcaaaagt gctgcagagt tagagaagct
     2161 gatgcctgta tctgcccaaa cgccaaaggg ccggaggttg agcggggaag agagaggcct
     2221 ctggtccacg gactcagccg aggaggacaa agaaaccaaa agaaatgaat ccaaagaaaa
     2281 atatcagaaa aggcatgatt ctgacaagga agaaaaaggc agaaaagagc caaaaggatt
     2341 aaagacactt aaggaaatca gaaatgcatt tgatttattt aaattaactc cagaagaaaa
     2401 aaatgatgtt tctgagaata atcggaaaag ggaagaaata ccactggatt ttaaaaccat
     2461 agacgatcac aaaaccaagg aaaacaaaca gtcacttaaa gaaaggagaa acaccagaga
     2521 cgaaacggat acttgggcat acattgctgc agaaggtgat caggaggttt tagacagcgt
     2581 gtgccaagca gatgagaatt cagatggcag gcagcagatt ctgagtttgg gcatggacct
     2641 gcagttggaa tggatgaagt tggaagattt ccaaaagcac cttgatggga aagatgagaa
     2701 ttttgctgca acagatgcaa ttccaagtaa tgtgttaagg gatgctgtga aaaatgggga
     2761 ttatattact gtaaaagttg cacttaattc aaatgaagaa tataacctgg accaagagga
     2821 ttccagtgga atgacactgg tgatgcttgc cgccgccgga gggcaggacg acctcctgcg
     2881 actcctcatc acaaaaggcg cgaaagtgaa cggtcggcag aagaacggga ccaccgccct
     2941 cattcatgct gcagagaaga actttttaac aacagtggct attcttttgg aagcaggagc
     3001 ttttgtaaat gtccagcaaa gcaatggtga gactgcactg atgaaggcct gtaaaagagg
     3061 aaattcagac atcgtacgac tcgtaattga atgtggagct gactgcaata ttttgtcaaa
     3121 gcaccagaat agtgccctgc actttgcgaa gcagtctaac aatgtgcttg tgtacgactt
     3181 gctgaagaac catttagaga cactttcaag agtagcagaa gagacaataa aggattactt
     3241 tgaagctcgc cttgctctgc tagaaccagt ttttccaatc gcatgtcatc gactctgtga
     3301 gggtccagat ttttcaacag atttcaatta caaaccccca cagaacatac cagaaggctc
     3361 tggcatcctg ctgtttatct tccatgcaaa ctttttgggt aaagaagtta ttgctcggct
     3421 ctgtggaccg tgtagtgtac aagctgtagt tctgaatgat aaatttcagc ttcctgtttt
     3481 tctggacagt cattttgttt actcattcag ccctgttgca ggtcccaata aactcttcat
     3541 aaggttgaca gaagcaccct ctgccaaggt taagttgcta ataggtgcat acagagtgca
     3601 gctgcagggg ggtggaggct ctcaccatca ccaccatcat caccaccatc actaaacggc
     3661 cggccgcggt catagctgtt tcctgaacag atcccgggtg gcatccctgt gacccctccc
     3721 cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa
     3781 aattaagttg catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg
     3841 gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt
     3901 gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg
     3961 ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca
     4021 ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg
     4081 tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac
     4141 aggcgtgaac cactgctccc ttccctgtcc ttctgatttt aaaataacta taccagcagg
     4201 aggacgtcca gacacagcat aggctacctg gccatgccca accggtggga catttgagtt
     4261 gcttgcttgg cactgtcctc tcatgcgttg ggtccactca gtagatgcct gttgaattgg
     4321 gtacgcggcc agcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4381 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4441 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4501 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4561 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4621 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4681 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4741 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4801 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4861 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4921 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4981 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5041 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5101 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5161 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta acctgaggct
     5221 atggcagggc ctgccgcccc gacgttggct gcgagccctg ggccttcacc cgaacttggg
     5281 gggtggggtg gggaaaagga agaaacgcgg gcgtattggc cccaatgggg tctcggtggg
     5341 gtatcgacag agtgccagcc ctgggaccga accccgcgtt tatgaacaaa cgacccaaca
     5401 ccgtgcgttt tattctgtct ttttattgcc gtcatagcgc gggttccttc cggtattgtc
     5461 tccttccgtg tttcagttag cctcccccta gggtgggcga agaactccag catgagatcc
     5521 ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca
     5581 tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc
     5641 atttcgaacc ccagagtccc gctcagaaga actcgtcaag aaggcgatag aaggcgatgc
     5701 gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc
     5761 caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcga tccgccacac
     5821 ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca
     5881 agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgctc gccttgagcc
     5941 tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga
     6001 caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga
     6061 atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata
     6121 ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata
     6181 gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg
     6241 tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca
     6301 ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat
     6361 cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg
     6421 ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct catcctgtct
     6481 cttgatcgat ctttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat
     6541 agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag tcagccatgg
     6601 ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg
     6661 actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
     6721 ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct
     6781 gctggggagc ctggggactt tccacaccct aactgacaca cattccacag ctggttcttt
     6841 ccgcctcagg actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6901 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6961 ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     7021 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     7081 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     7141 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     7201 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     7261 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     7321 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     7381 agctgattta acaaaaattt aacgcgaatt tt
//
