LOCUS       sequence-173855-        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"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             553..1227
                     /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(683..701)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(733..753)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     951..970
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1145..1168
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    1243..1267
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3887..3911)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    4087..4208
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4124..4143)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4178..4197
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4215..4670)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4352..4373)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4564..4583
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4697..4801
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4803..5160
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4803..4994
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4826..4846)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5011..5146
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5195..5989
                     /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             5195..5989
                     /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(5249..5268)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5859..5878
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6177..6196)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6221..6268
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6597..7185
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7086..7105
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggcctc ctccgaggac gtcatcaagg agttcatgcg cttcaaggtg
      601 cgcatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
      661 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggcggccc cctgcccttc
      721 gcctgggaca tcctgtcccc ccagttccag tacggctcca aggtgtacgt gaagcacccc
      781 gccgacatcc ccgactacaa gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
      841 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
      901 tccttcatct acaaggtgaa gttcatcggc gtgaacttcc cctccgacgg ccccgtaatg
      961 cagaagaaga ctatgggctg ggaggcctcc accgagcgcc tgtacccccg cgacggcgtg
     1021 ctgaagggcg agatccacaa ggccctgaag ctgaaggacg gcggccacta cctggtggag
     1081 ttcaagtcca tctacatggc caagaagccc gtgcagctgc ccggctacta ctacgtggac
     1141 tccaagctgg acatcacctc ccacaacgag gactacacca tcgtggagca gtacgagcgc
     1201 gccgagggcc gccaccacct gttcctgaga tctcgagatc aaacaagttt gtacaaaaaa
     1261 gcaggcttca ccatgacagc tgacaaggag aagaaaagga gtagctcgga gaggaggaag
     1321 gagaagtccc gggatgctgc gcggtgccgg cggagcaagg agacggaggt gttctatgag
     1381 ctggcccatg agctgcctct gccccacagt gtgagctccc atctggacaa ggcctccatc
     1441 atgcgactgg caatcagctt cctgcgaaca cacaagctcc tctcctcagt ttgctctgaa
     1501 aacgagtccg aagccgaagc tgaccagcag atggacaact tgtacctgaa agccttggag
     1561 ggtttcattg ccgtggtgac ccaagatggc gacatgatct ttctgtcaga aaacatcagc
     1621 aagttcatgg gacttacaca ggtggagcta acaggacata gtatctttga cttcactcat
     1681 ccctgcgacc atgaggagat tcgtgagaac ctgagtctca aaaatggctc tggttttggg
     1741 aaaaaaagca aagacatgtc cacagagcgg gacttcttca tgaggatgaa gtgcacggtc
     1801 accaacagag gccgtactgt caacctcaag tcagccacct ggaaggtctt gcactgcacg
     1861 ggccaggtga aagtctacaa caactgccct cctcacaata gtctgtgtgg ctacaaggag
     1921 cccctgctgt cctgcctcat catcatgtgt gaaccaatcc agcacccatc ccacatggac
     1981 atccccctgg atagcaagac cttcctgagc cgccacagca tggacatgaa gttcacctac
     2041 tgtgatgaca gaatcacaga actgattggt taccaccctg aggagctgct tggccgctca
     2101 gcctatgaat tctaccatgc gctagactcc gagaacatga ccaagagtca ccagaacttg
     2161 tgcaccaagg gtcaggtagt aagtggccag taccggatgc tcgcaaagca tgggggctac
     2221 gtgtggctgg agacccaggg gacggtcatc tacaaccctc gcaacctgca gccccagtgc
     2281 atcatgtgtg tcaactacgt cctgagtgag attgagaaga atgacgtggt gttctccatg
     2341 gaccagactg aatccctgtt caagccccac ctgatggcca tgaacagcat ctttgatagc
     2401 agtggcaagg gggctgtgtc tgagaagagt aacttcctat tcaccaagct aaaggaggag
     2461 cccgaggagc tggcccagct ggctcccacc ccaggagacg ccatcatctc tctggatttc
     2521 gggaatcaga acttcgagga gtcctcagcc tatggcaagg ccatcctgcc cccgagccag
     2581 ccatgggcca cggagttgag gagccacagc acccagagcg aggctgggag cctgcctgcc
     2641 ttcaccgtgc cccaggcagc tgccccgggc agcaccaccc ccagtgccac cagcagcagc
     2701 agcagctgct ccacgcccaa tagccctgaa gactattaca catctttgga taacgacctg
     2761 aagattgaag tgattgagaa gctcttcgcc atggacacag aggccaagga ccaatgcagt
     2821 acccagacgg atttcaatga gctggacttg gagacactgg caccctatat ccccatggac
     2881 ggggaagact tccagctaag ccccatctgc cccgaggagc ggctcttggc ggagaaccca
     2941 cagtccaccc cccagcactg cttcagtgcc atgacaaaca tcttccagcc actggcccct
     3001 gtagccccgc acagtccctt cctcctggac aagtttcagc agcagctgga gagcaagaag
     3061 acagagcccg agcaccggcc catgtcctcc atcttctttg atgccggaag caaagcatcc
     3121 ctgccaccgt gctgtggcca ggccagcacc cctctctctt ccatgggggg cagatccaat
     3181 acccagtggc ccccagatcc accattacat tttgggccca caaagtgggc cgtcggggat
     3241 cagcgcacag agttcttggg agcagcgccg ttggggcccc ctgtctctcc accccatgtc
     3301 tccaccttca agacaaggtc tgcaaagggt tttggggctc gaggcccaga cgtgctgagt
     3361 ccggccatgg tagccctctc caacaagctg aagctgaagc gacagctgga gtatgaagag
     3421 caagccttcc aggacctgag cgggggggac ccacctggtg gcagcacctc acatttgatg
     3481 tggaaacgga tgaagaacct caggggtggg agctgccctt tgatgccgga caagccactg
     3541 agcgcaaatg tacccaatga taagttcacc caaaacccca tgaggggcct gggccatccc
     3601 ctgagacatc tgccgctgcc acagcctcca tctgccatca gtcccgggga gaacagcaag
     3661 agcaggttcc ccccacagtg ctacgccacc cagtaccagg actacagcct gtcgtcagcc
     3721 cacaaggtgt caggcatggc aagccggctg ctcgggccct catttgagtc ctacctgctg
     3781 cccgaactga ccagatatga ctgtgaggtg aacgtgcccg tgctgggaag ctccacgctc
     3841 ctgcaaggag gggacctcct cagagccctg gacgaggcca cctgagaccc agctttcttg
     3901 tacaaagtgg ttcgatctca agcttcgaat tctgcagtcg acggtaccgc gggcccggga
     3961 tccaccggat ctagataact gatcataatc agccatacca catttgtaga ggttttactt
     4021 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     4081 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4141 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4201 gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     4261 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     4321 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     4381 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     4441 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     4501 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     4561 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     4621 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     4681 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4741 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4801 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4861 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4921 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4981 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     5041 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     5101 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     5161 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     5221 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     5281 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     5341 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     5401 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     5461 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     5521 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     5581 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5641 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5701 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5761 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5821 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5881 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5941 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     6001 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     6061 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     6121 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     6181 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     6241 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     6301 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     6361 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     6421 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     6481 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6541 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6601 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6661 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6721 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6781 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6841 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6901 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6961 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     7021 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7081 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7141 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     7201 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     7261 tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat
     7321 tacggggtca ttagttcata gcccatatat ggagttccg
//
