LOCUS       sequence-382111-        6576 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses first 1200 nt of lncRNA HIF1a-AS1 (unspliced) .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6576)
  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 6576)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pcDNA3.1+HIF1a-AS1_1200_unspliced
FEATURES             Location/Qualifiers
     source          1..6576
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2167..2184)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2173..2397
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2443..2871
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2530..2549)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2740..2761
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2880..2900)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2885..3214
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3065..3200
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3281..4075
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3335..3354)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3945..3964
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4249..4370
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4286..4305)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4340..4359
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4419..4435)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4419..4435)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4432..4454)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4443..4459
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4467..4473,4474..4491,4492..4497))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4512..4533
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4650..4667)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4821..5409)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4901..4920)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5580..6371,6372..6440))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6203..6222
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6441..6545)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta cccgccgccg gcgccctcca tggtgaatcg gtccccgcga
      961 tgtcttcacg gcgggcggcc cccaggctcg ctccggccta agcgctggct ccctccacac
     1021 gcggagaaga gaaggaaagg caagtccaga ggtgggggtg cgaggcggga aacccctcgt
     1081 gagactagag agaagcgggc ggcaatcgcc ggggtccggg cccggctccg agcctctcct
     1141 caggtggctt gtcagggccc agcccgacac actggccgaa gcgacgaaga gggtgatcct
     1201 cctgtcccct cagacgaggc agcactgtgc actgaggagc tgaggcagcg tcagggggcg
     1261 ggcaagggcg gaggcgcgct cgggcgcgcg gggaggggag agggggcggg agcgcgcccg
     1321 ctcgcctgcc caggccggcg cgcgctcccg caagcccgcc tcacctgagg tggaggcggg
     1381 ttcctcgaga tccaatggcg agcgacgcag gcggcaagtc cccaccccag cccacgccgc
     1441 tctcagccaa tcaggaggcg gtcagctgag gcacagctgg gacgggctgc gacgctcacg
     1501 tgctcgtctg tgtttagcgg cggaggaaag agaaaggaga tgggggtgcg gctcagcccg
     1561 agtctgggga ctctgcgcct tctccgaagg aaggcggtgc ccggctttgg gaggcgctgc
     1621 tgagaaggga tttcggttgc ctggtcccaa acatgcatca tattttccta ctcttggtgc
     1681 agtaatgtaa tgctcctgct ctctttctcc tcggcctcag tgctgggcac gattgccgcc
     1741 caactctgct gggctctctg ttcaaaaaag cagacttcgc ctcgtggcag caagcccttc
     1801 ctttggtctc gttgctcaga tgtgttacaa cggggtcttt ccttaccaga ttacctaacc
     1861 atgggtcaat gtcacccacc gcactctttg ccacggagca caggggaact caccttgtct
     1921 actaagtttt tctgtggcgg agtttactta ctgaataaat gcaattaaaa ataaaaatga
     1981 actcctgttg ctgtaaactt caagggaaat aaagttgcag ggaaatctta aaatctgaaa
     2041 ctatgttaag catttgcaaa taggtataaa tgactttaat aagcatataa accaatttca
     2101 gggaaagagt agaaaaaatg gcgcggccgc tcgagtctag agggcccgtt taaacccgct
     2161 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     2221 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2281 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     2341 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     2401 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     2461 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     2521 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2581 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     2641 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     2701 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     2761 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     2821 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     2881 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2941 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3001 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3061 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3121 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     3181 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     3241 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3301 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3361 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3421 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     3481 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3541 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3601 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3661 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     3721 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     3781 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     3841 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     3901 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     3961 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4021 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4081 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4141 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4201 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     4261 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4321 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     4381 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     4441 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4501 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4561 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4621 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4681 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4741 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4801 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4861 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4921 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4981 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5041 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5101 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5161 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5221 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5281 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5341 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5401 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5461 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5521 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5581 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5641 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5701 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5761 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5821 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5881 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5941 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6001 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6061 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6121 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6181 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6241 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6301 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6361 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6421 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6481 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6541 ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
