LOCUS       sequence-382027-        6155 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MPP8-delta442-860_10xHis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6155)
  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 6155)
  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-delta442-860-10xHis
FEATURES             Location/Qualifiers
     source          1..6155
                     /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             2366..2395
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(2412..2428)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2412..2428)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2439..3061
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2638..2657)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(3210..3798)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3290..3309)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4127..4174
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4199..4218
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4406..5200)
                     /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(4406..5200)
                     /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(4517..4536)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5127..5146
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5235..5592)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5249..5384
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5401..5592
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5549..5569
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(5594..5698)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5725..6153
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5812..5831)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6022..6043
                     /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 tcgctggaaa 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 gggggtggag gctctcacca tcaccaccat catcaccacc atcactaaac
     2401 ggccggccgc ggtcatagct gtttcctgaa cagatcccgg gtggcatccc tgtgacccct
     2461 ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa
     2521 taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga
     2581 ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct
     2641 attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct
     2701 gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga
     2761 ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc
     2821 tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat
     2881 tacaggcgtg aaccactgct cccttccctg tccttctgat tttaaaataa ctataccagc
     2941 aggaggacgt ccagacacag cataggctac ctggccatgc ccaaccggtg ggacatttga
     3001 gttgcttgct tggcactgtc ctctcatgcg ttgggtccac tcagtagatg cctgttgaat
     3061 tgggtacgcg gccagcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3121 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3181 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3241 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3301 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3361 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3421 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3481 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3541 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3601 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3661 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3721 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3781 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3841 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3901 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaacctgag
     3961 gctatggcag ggcctgccgc cccgacgttg gctgcgagcc ctgggccttc acccgaactt
     4021 ggggggtggg gtggggaaaa ggaagaaacg cgggcgtatt ggccccaatg gggtctcggt
     4081 ggggtatcga cagagtgcca gccctgggac cgaaccccgc gtttatgaac aaacgaccca
     4141 acaccgtgcg ttttattctg tctttttatt gccgtcatag cgcgggttcc ttccggtatt
     4201 gtctccttcc gtgtttcagt tagcctcccc ctagggtggg cgaagaactc cagcatgaga
     4261 tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa gcccaacctt
     4321 tcatagaagg cggcggtgga atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg
     4381 gtcatttcga accccagagt cccgctcaga agaactcgtc aagaaggcga tagaaggcga
     4441 tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc
     4501 cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag cgatccgcca
     4561 cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg
     4621 gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg ctcgccttga
     4681 gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat
     4741 cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt
     4801 cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg
     4861 atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca
     4921 atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc
     4981 ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg cagttcattc agggcaccgg
     5041 acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg
     5101 catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag
     5161 cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg
     5221 tctcttgatc gatctttgca aaagcctagg cctccaaaaa agcctcctca ctacttctgg
     5281 aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca
     5341 tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg agttaggggc
     5401 gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag
     5461 cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg catacttctg
     5521 cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca cagctggttc
     5581 tttccgcctc aggactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5641 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5701 catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     5761 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     5821 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     5881 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     5941 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     6001 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     6061 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     6121 atgagctgat ttaacaaaaa tttaacgcga atttt
//
