LOCUS       sequence-382025-        6992 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MPP8-delta600-728_10xHis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6992)
  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 6992)
  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-delta600-728-10xHis
FEATURES             Location/Qualifiers
     source          1..6992
                     /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             3236..3265
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(3282..3298)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3282..3298)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3309..3931
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3508..3527)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     rep_origin      complement(4080..4668)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4160..4179)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4997..5044
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5069..5088
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5276..6070)
                     /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(5276..6070)
                     /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(5387..5406)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5997..6016
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     enhancer        6238..6429
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6386..6406
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(6431..6535)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6562..6990
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6649..6668)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6859..6880
                     /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 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 ttccgtagca gaagagacaa taaaggatta ctttgaagct cgccttgctc tgctagaacc
     2881 agtttttcca atcgcatgtc atcgactctg tgagggtcca gatttttcaa cagatttcaa
     2941 ttacaaaccc ccacagaaca taccagaagg ctctggcatc ctgctgttta tcttccatgc
     3001 aaactttttg ggtaaagaag ttattgctcg gctctgtgga ccgtgtagtg tacaagctgt
     3061 agttctgaat gataaatttc agcttcctgt ttttctggac agtcattttg tttactcatt
     3121 cagccctgtt gcaggtccca ataaactctt cataaggttg acagaagcac cctctgccaa
     3181 ggttaagttg ctaataggtg catacagagt gcagctgcag gggggtggag gctctcacca
     3241 tcaccaccat catcaccacc atcactaaac ggccggccgc ggtcatagct gtttcctgaa
     3301 cagatcccgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg
     3361 ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact
     3421 aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg
     3481 ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac
     3541 aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc
     3601 ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt
     3661 agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct
     3721 acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg
     3781 tccttctgat tttaaaataa ctataccagc aggaggacgt ccagacacag cataggctac
     3841 ctggccatgc ccaaccggtg ggacatttga gttgcttgct tggcactgtc ctctcatgcg
     3901 ttgggtccac tcagtagatg cctgttgaat tgggtacgcg gccagcggcg agcggtatca
     3961 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4021 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4081 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4141 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4201 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4261 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4321 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4381 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4441 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4501 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4561 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4621 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4681 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4741 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4801 tcaatctaaa gtatatatga gtaacctgag gctatggcag ggcctgccgc cccgacgttg
     4861 gctgcgagcc ctgggccttc acccgaactt ggggggtggg gtggggaaaa ggaagaaacg
     4921 cgggcgtatt ggccccaatg gggtctcggt ggggtatcga cagagtgcca gccctgggac
     4981 cgaaccccgc gtttatgaac aaacgaccca acaccgtgcg ttttattctg tctttttatt
     5041 gccgtcatag cgcgggttcc ttccggtatt gtctccttcc gtgtttcagt tagcctcccc
     5101 ctagggtggg cgaagaactc cagcatgaga tccccgcgct ggaggatcat ccagccggcg
     5161 tcccggaaaa cgattccgaa gcccaacctt tcatagaagg cggcggtgga atcgaaatct
     5221 cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga accccagagt cccgctcaga
     5281 agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt
     5341 aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag
     5401 ccaacgctat gtcctgatag cgatccgcca cacccagccg gccacagtcg atgaatccag
     5461 aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga
     5521 gatcctcgcc gtcgggcatg ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc
     5581 cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg
     5641 ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat
     5701 gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg
     5761 acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga
     5821 caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg
     5881 cctcgtcttg cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc
     5941 gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc
     6001 agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt
     6061 gttcaatcat gcgaaacgat cctcatcctg tctcttgatc gatctttgca aaagcctagg
     6121 cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc
     6181 tctgcataaa taaaaaaaat tagtcagcca tggggcggga tgggcggagt taggggcggg
     6241 actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
     6301 ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct
     6361 gctggggagc ctggggactt tccacaccct aactgacaca cattccacag ctggttcttt
     6421 ccgcctcagg actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6481 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6541 ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     6601 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     6661 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     6721 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     6781 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     6841 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     6901 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     6961 agctgattta acaaaaattt aacgcgaatt tt
//
