LOCUS       sequence-382026-        7220 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MPP8-delta55-118_10xHis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7220)
  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 7220)
  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-delta55-118-10xHis
FEATURES             Location/Qualifiers
     source          1..7220
                     /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             3431..3460
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(3477..3493)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3477..3493)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3504..4126
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3703..3722)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(4275..4863)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4355..4374)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5192..5239
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5264..5283
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5471..6265)
                     /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(5471..6265)
                     /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(5582..5601)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6192..6211
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(6300..6657)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6314..6449
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6466..6657
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6614..6634
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(6659..6763)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6790..7218
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6877..6896)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7087..7108
                     /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 gaggaggaca ggaaggatat
     1201 tcagagacta tccttaaata acgacatatt tgaggcgaac tctgatagcg atcagcaaag
     1261 tgagacaaaa gaagatactt ccccaaagaa gaaaaagaaa aaattgaggc agagagaaga
     1321 gaaaagccca gatgatctga aaaagaaaaa agcaaaggcc gggaagctaa aagacaagtc
     1381 caaaccagac ctggagagct ccttggaaag tttagttttt gatttaagga caaagaaaag
     1441 aatttctgaa gccaaagaag aactaaagga gtccaaaaag cccaaaaaag atgaagtaaa
     1501 agaaacaaaa gaattaaaga aagttaaaaa gggtgaaata agagatttaa agacgaaaac
     1561 aagagaagat cccaaagaaa atagaaaaac aaaaaaagaa aaatttgtcg aatcccaggt
     1621 ggaatctgaa tcaagtgtac ttaatgattc tccctttcca gaggatgaca gtgaagggct
     1681 acattccgac agcagagaag agaaacaaaa cactaaaagt gcaagagaga gagcagggca
     1741 ggacatgggg ctggagcatg gctttgagaa gcccctagac agtgccatga gtgctgagga
     1801 ggataccgat gtcagaggca ggaggaaaaa gaagaccccg agaaaggctg aggacactag
     1861 agagaacagg aagctagaga acaagaacgc tttcttagag aagaaaactg tgcctaaaaa
     1921 gcagaggaat caagacagaa gcaaaagtgc tgcagagtta gagaagctga tgcctgtatc
     1981 tgcccaaacg ccaaagggcc ggaggttgag cggggaagag agaggcctct ggtccacgga
     2041 ctcagccgag gaggacaaag aaaccaaaag aaatgaatcc aaagaaaaat atcagaaaag
     2101 gcatgattct gacaaggaag aaaaaggcag aaaagagcca aaaggattaa agacacttaa
     2161 ggaaatcaga aatgcatttg atttatttaa attaactcca gaagaaaaaa atgatgtttc
     2221 tgagaataat cggaaaaggg aagaaatacc actggatttt aaaaccatag acgatcacaa
     2281 aaccaaggaa aacaaacagt cacttaaaga aaggagaaac accagagacg aaacggatac
     2341 ttgggcatac attgctgcag aaggtgatca ggaggtttta gacagcgtgt gccaagcaga
     2401 tgagaattca gatggcaggc agcagattct gagtttgggc atggacctgc agttggaatg
     2461 gatgaagttg gaagatttcc aaaagcacct tgatgggaaa gatgagaatt ttgctgcaac
     2521 agatgcaatt ccaagtaatg tgttaaggga tgctgtgaaa aatggggatt atattactgt
     2581 aaaagttgca cttaattcaa atgaagaata taacctggac caagaggatt ccagtggaat
     2641 gacactggtg atgcttgccg ccgccggagg gcaggacgac ctcctgcgac tcctcatcac
     2701 aaaaggcgcg aaagtgaacg gtcggcagaa gaacgggacc accgccctca ttcatgctgc
     2761 agagaagaac tttttaacaa cagtggctat tcttttggaa gcaggagctt ttgtaaatgt
     2821 ccagcaaagc aatggtgaga ctgcactgat gaaggcctgt aaaagaggaa attcagacat
     2881 cgtacgactc gtaattgaat gtggagctga ctgcaatatt ttgtcaaagc accagaatag
     2941 tgccctgcac tttgcgaagc agtctaacaa tgtgcttgtg tacgacttgc tgaagaacca
     3001 tttagagaca ctttcaagag tagcagaaga gacaataaag gattactttg aagctcgcct
     3061 tgctctgcta gaaccagttt ttccaatcgc atgtcatcga ctctgtgagg gtccagattt
     3121 ttcaacagat ttcaattaca aacccccaca gaacatacca gaaggctctg gcatcctgct
     3181 gtttatcttc catgcaaact ttttgggtaa agaagttatt gctcggctct gtggaccgtg
     3241 tagtgtacaa gctgtagttc tgaatgataa atttcagctt cctgtttttc tggacagtca
     3301 ttttgtttac tcattcagcc ctgttgcagg tcccaataaa ctcttcataa ggttgacaga
     3361 agcaccctct gccaaggtta agttgctaat aggtgcatac agagtgcagc tgcagggggg
     3421 tggaggctct caccatcacc accatcatca ccaccatcac taaacggccg gccgcggtca
     3481 tagctgtttc ctgaacagat cccgggtggc atccctgtga cccctcccca gtgcctctcc
     3541 tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca
     3601 tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga
     3661 gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct
     3721 ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc
     3781 tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat
     3841 ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct
     3901 aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca
     3961 ctgctccctt ccctgtcctt ctgattttaa aataactata ccagcaggag gacgtccaga
     4021 cacagcatag gctacctggc catgcccaac cggtgggaca tttgagttgc ttgcttggca
     4081 ctgtcctctc atgcgttggg tccactcagt agatgcctgt tgaattgggt acgcggccag
     4141 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4201 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4261 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4321 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4381 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4441 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4501 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4561 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4621 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4681 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4741 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4801 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4861 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4921 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4981 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaac ctgaggctat ggcagggcct
     5041 gccgccccga cgttggctgc gagccctggg ccttcacccg aacttggggg gtggggtggg
     5101 gaaaaggaag aaactcgggc gtattggccc caatggggtc tcggtggggt atcgacagag
     5161 tgccagccct gggaccgaac cccgcgttta tgaacaaacg acccaacacc gtgcgtttta
     5221 ttctgtcttt ttattgccgt catagcgcgg gttccttccg gtattgtctc cttccgtgtt
     5281 tcagttagcc tccccctagg gtgggcgaag aactccagca tgagatcccc gcgctggagg
     5341 atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg
     5401 gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc
     5461 agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
     5521 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag
     5581 caatatcacg ggtagccaac gctatgtcct gatagcgatc cgccacaccc agccggccac
     5641 agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc
     5701 catgggtcac gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg gcgaacagtt
     5761 cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt
     5821 ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag
     5881 ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag
     5941 gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc
     6001 ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc
     6061 acgatagccg cgctgcctcg tcttgcagtt cattcagggc accggacagg tcggtcttga
     6121 caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga
     6181 ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg
     6241 cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcgatct
     6301 ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc
     6361 gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg
     6421 gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct
     6481 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
     6541 cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
     6601 ggggactttc cacaccctaa ctgacacaca ttccacagct ggttctttcc gcctcaggac
     6661 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6721 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6781 tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     6841 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     6901 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     6961 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     7021 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     7081 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     7141 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     7201 aaaaatttaa cgcgaatttt
//
