LOCUS       sequence-170216-        7359 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminally dsRedXP-tagged human HIF-2a/EPAS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7359)
  AUTHORS   Taylor SE, Bagnall J, Mason D, Levy R, Fernig DG, See V
  TITLE     Differential sub-nuclear distribution of hypoxia-inducible factors 
            (HIF)-1 and -2 alpha impacts on their stability and mobility.
  JOURNAL   Open Biol. 2016 Sep;6(9). pii: 160195.
  PUBMED    27655733
REFERENCE   2  (bases 1 to 7359)
  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: "Open 
            Biol."; date: "2016-09"; volume: "6(9). pii"; pages: " 160195"
COMMENT     Sequence Label: pg-dsRedXP-HIF-2alpha
FEATURES             Location/Qualifiers
     source          1..7359
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    190..311
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(227..246)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     281..300
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(318..773)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(455..476)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     667..686
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        800..904
                     /gene="bla"
                     /label=AmpR promoter
     promoter        906..1263
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        906..1097
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(929..949)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1114..1249
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1298..2092
                     /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             1298..2092
                     /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(1352..1371)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1962..1981
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2280..2299)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2324..2371
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2700..3288
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3189..3208
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3463..3766
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3767..3970
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3921..3941
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4009..4018
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4015..4689
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(4145..4163)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(4195..4215)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     4413..4432
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4607..4630
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    4705..4729
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(join(7349..7359,1..14))
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 ttgtacaaag tggttcgatc tcaagcttcg aattctgcag tcgacggtac cgcgggcccg
       61 ggatccaccg gatctagata actgatcata atcagccata ccacatttgt agaggtttta
      121 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
      181 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
      241 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
      301 aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
      361 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
      421 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
      481 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
      541 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
      601 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
      661 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
      721 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
      781 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
      841 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
      901 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
      961 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     1021 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     1081 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     1141 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     1201 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     1261 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     1321 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     1381 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     1441 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     1501 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     1561 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     1621 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     1681 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     1741 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     1801 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     1861 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     1921 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     1981 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     2041 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     2101 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     2161 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     2221 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
     2281 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     2341 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     2401 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     2461 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     2521 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
     2581 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     2641 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2701 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2761 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2821 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     2881 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2941 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3001 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3061 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3121 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     3181 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     3241 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     3301 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     3361 ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc
     3421 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     3481 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     3541 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     3601 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     3661 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     3721 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     3781 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     3841 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     3901 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     3961 aagcagagct ggtttagtga accgtcagat ccgctagcgc taccggtcgc caccatggcc
     4021 tcctccgagg acgtcatcaa ggagttcatg cgcttcaagg tgcgcatgga gggctccgtg
     4081 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     4141 accgccaagc tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc
     4201 ccccagttcc agtacggctc caaggtgtac gtgaagcacc ccgccgacat ccccgactac
     4261 aagaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     4321 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gctccttcat ctacaaggtg
     4381 aagttcatcg gcgtgaactt cccctccgac ggccccgtaa tgcagaagaa gactatgggc
     4441 tgggaggcct ccaccgagcg cctgtacccc cgcgacggcg tgctgaaggg cgagatccac
     4501 aaggccctga agctgaagga cggcggccac tacctggtgg agttcaagtc catctacatg
     4561 gccaagaagc ccgtgcagct gcccggctac tactacgtgg actccaagct ggacatcacc
     4621 tcccacaacg aggactacac catcgtggag cagtacgagc gcgccgaggg ccgccaccac
     4681 ctgttcctga gatctcgaga tcaaacaagt ttgtacaaaa aagcaggctt caccatgaca
     4741 gctgacaagg agaagaaaag gagtagctcg gagaggagga aggagaagtc ccgggatgct
     4801 gcgcggtgcc ggcggagcaa ggagacggag gtgttctatg agctggccca tgagctgcct
     4861 ctgccccaca gtgtgagctc ccatctggac aaggcctcca tcatgcgact ggcaatcagc
     4921 ttcctgcgaa cacacaagct cctctcctca gtttgctctg aaaacgagtc cgaagccgaa
     4981 gctgaccagc agatggacaa cttgtacctg aaagccttgg agggtttcat tgccgtggtg
     5041 acccaagatg gcgacatgat ctttctgtca gaaaacatca gcaagttcat gggacttaca
     5101 caggtggagc taacaggaca tagtatcttt gacttcactc atccctgcga ccatgaggag
     5161 attcgtgaga acctgagtct caaaaatggc tctggttttg ggaaaaaaag caaagacatg
     5221 tccacagagc gggacttctt catgaggatg aagtgcacgg tcaccaacag aggccgtact
     5281 gtcaacctca agtcagccac ctggaaggtc ttgcactgca cgggccaggt gaaagtctac
     5341 aacaactgcc ctcctcacaa tagtctgtgt ggctacaagg agcccctgct gtcctgcctc
     5401 atcatcatgt gtgaaccaat ccagcaccca tcccacatgg acatccccct ggatagcaag
     5461 accttcctga gccgccacag catggacatg aagttcacct actgtgatga cagaatcaca
     5521 gaactgattg gttaccaccc tgaggagctg cttggccgct cagcctatga attctaccat
     5581 gcgctagact ccgagaacat gaccaagagt caccagaact tgtgcaccaa gggtcaggta
     5641 gtaagtggcc agtaccggat gctcgcaaag catgggggct acgtgtggct ggagacccag
     5701 gggacggtca tctacaaccc tcgcaacctg cagccccagt gcatcatgtg tgtcaactac
     5761 gtcctgagtg agattgagaa gaatgacgtg gtgttctcca tggaccagac tgaatccctg
     5821 ttcaagcccc acctgatggc catgaacagc atctttgata gcagtggcaa gggggctgtg
     5881 tctgagaaga gtaacttcct attcaccaag ctaaaggagg agcccgagga gctggcccag
     5941 ctggctccca ccccaggaga cgccatcatc tctctggatt tcgggaatca gaacttcgag
     6001 gagtcctcag cctatggcaa ggccatcctg cccccgagcc agccatgggc cacggagttg
     6061 aggagccaca gcacccagag cgaggctggg agcctgcctg ccttcaccgt gccccaggca
     6121 gctgccccgg gcagcaccac ccccagtgcc accagcagca gcagcagctg ctccacgccc
     6181 aatagccctg aagactatta cacatctttg gataacgacc tgaagattga agtgattgag
     6241 aagctcttcg ccatggacac agaggccaag gaccaatgca gtacccagac ggatttcaat
     6301 gagctggact tggagacact ggcaccctat atccccatgg acggggaaga cttccagcta
     6361 agccccatct gccccgagga gcggctcttg gcggagaacc cacagtccac cccccagcac
     6421 tgcttcagtg ccatgacaaa catcttccag ccactggccc ctgtagcccc gcacagtccc
     6481 ttcctcctgg acaagtttca gcagcagctg gagagcaaga agacagagcc cgagcaccgg
     6541 cccatgtcct ccatcttctt tgatgccgga agcaaagcat ccctgccacc gtgctgtggc
     6601 caggccagca cccctctctc ttccatgggg ggcagatcca atacccagtg gcccccagat
     6661 ccaccattac attttgggcc cacaaagtgg gccgtcgggg atcagcgcac agagttcttg
     6721 ggagcagcgc cgttggggcc ccctgtctct ccaccccatg tctccacctt caagacaagg
     6781 tctgcaaagg gttttggggc tcgaggccca gacgtgctga gtccggccat ggtagccctc
     6841 tccaacaagc tgaagctgaa gcgacagctg gagtatgaag agcaagcctt ccaggacctg
     6901 agcggggggg acccacctgg tggcagcacc tcacatttga tgtggaaacg gatgaagaac
     6961 ctcaggggtg ggagctgccc tttgatgccg gacaagccac tgagcgcaaa tgtacccaat
     7021 gataagttca cccaaaaccc catgaggggc ctgggccatc ccctgagaca tctgccgctg
     7081 ccacagcctc catctgccat cagtcccggg gagaacagca agagcaggtt ccccccacag
     7141 tgctacgcca cccagtacca ggactacagc ctgtcgtcag cccacaaggt gtcaggcatg
     7201 gcaagccggc tgctcgggcc ctcatttgag tcctacctgc tgcccgaact gaccagatat
     7261 gactgtgagg tgaacgtgcc cgtgctggga agctccacgc tcctgcaagg aggggacctc
     7321 ctcagagccc tggaccaggc cacctgagac ccagctttc
//
