LOCUS       sequence-173858-        5169 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HIF-1a's ODD domain fused to EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5169)
  AUTHORS   Bagnall J, Leedale J, Taylor SE, Spiller DG, White MR, Sharkey KJ, 
            Bearon RN, See V
  TITLE     Tight control of hypoxia-inducible factor-alpha transient dynamics 
            is essential for cell survival in hypoxia.
  JOURNAL   J Biol Chem. 2014 Feb 28;289(9):5549-64. doi: 
            10.1074/jbc.M113.500405. Epub 2014 Jan 6.
  PUBMED    24394419
REFERENCE   2  (bases 1 to 5169)
  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"; doi: 
            "10.1074/jbc.M113.500405"; journalName: "J Biol Chem"; date: 
            "2014-02-28- 28"; volume: "289"; issue: "9"; pages: "5549-64"
COMMENT     Sequence Label: pg-ODD-EGFP
FEATURES             Location/Qualifiers
     source          1..5169
                     /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"
     protein_bind    563..587
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(966..990)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1049..1058
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1055..1057,1058..1060,1061..1774)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1055..1057,1058..1060,1061..1771)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1100..1121)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1361..1380)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1708..1729
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1897..2018
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1934..1953)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1988..2007
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2025..2480)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2162..2183)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2374..2393
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2507..2611
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2613..2970
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2613..2804
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2636..2656)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2821..2956
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3005..3799
                     /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             3005..3799
                     /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(3059..3078)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3669..3688
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3987..4006)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4031..4078
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4407..4995
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4896..4915
                     /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 ccggactcag atctcgagat caacaagttt gtacaaaaaa gcaggcttca ccatggaatt
      601 caagttggaa ttggtagaaa aactttttgc tgaagacaca gaagcaaaga acccattttc
      661 tactcaggac acagatttag acttggagat gttagctccc tatatcccaa tggatgatga
      721 cttccagtta cgttccttcg atcagttgtc accattagaa agcagttccg caagccctga
      781 aagcgcaagt cctcaaagca cagttacagt attccagcag actcaaatac aagaacctac
      841 tgctaatgcc accactacca ctgccaccac tgatgaatta aaaacagtga caaaagaccg
      901 tatggaagac attaaaatat tgattgcatc tccatctcct acccacatac ataaagaaac
      961 tactgaccca gctttcttgt acaaagtggt tgatctcaag cttcgaattc tgcagtcgac
     1021 ggtaccgcgg gcccgggatc caccggtcgc caccatggtg agcaagggcg aggagctgtt
     1081 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     1141 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     1201 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     1261 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     1321 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     1381 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1441 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     1501 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     1561 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     1621 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     1681 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     1741 gatcactctc ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc
     1801 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     1861 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     1921 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     1981 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
     2041 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     2101 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     2161 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     2221 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     2281 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     2341 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     2401 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     2461 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     2521 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     2581 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     2641 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2701 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     2761 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     2821 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     2881 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     2941 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     3001 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3061 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3121 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3181 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3241 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     3301 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3361 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     3421 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     3481 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     3541 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     3601 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     3661 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     3721 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     3781 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     3841 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     3901 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     3961 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     4021 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     4081 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     4141 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     4201 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     4261 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4321 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4381 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4441 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4501 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     4561 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4621 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4681 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4741 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4801 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4861 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     4921 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4981 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5041 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     5101 gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     5161 ggagttccg
//
