LOCUS       sequence-384561-        6359 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PHD2-EGFP expressed under the control of the PHD2 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6359)
  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 6359)
  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-proPHD2-PHD2-EGFP
FEATURES             Location/Qualifiers
     source          1..6359
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..147
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        164..355
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     312..332
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(357..461)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      488..943
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(575..594)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     785..806
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    950..1071
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(961..980)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1015..1034
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(1194..1907,1908..1910,1911..1913))
                     /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             complement(join(1197..1907,1908..1910,1911..1913))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1239..1260)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1588..1607
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1847..1868
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1910..1919
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3224..3233
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     rep_origin      complement(4332..4920)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4412..4431)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5249..5296
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5321..5340
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5528..6322)
                     /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(5528..6322)
                     /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(5639..5658)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6249..6268
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(6357..6359,1..355))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga
       61 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
      121 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
      181 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
      241 cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
      301 ggactttcca caccctaact gacacacatt ccacagctgg ttctttccgc ctcaggactc
      361 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
      421 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
      481 ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
      541 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
      601 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
      661 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
      721 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
      781 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
      841 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
      901 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttacgcctt aagatacatt
      961 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     1021 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     1081 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
     1141 taaaacctct acaaatgtgg tatggctgat tatgatctag agtcgcggcc gctttacttg
     1201 tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag caggaccatg
     1261 tgatcgcgct tctcgttggg gtctttgctc agggcggact gggtgctcag gtagtggttg
     1321 tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc
     1381 tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt tcaccttgat gccgttcttc
     1441 tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc
     1501 aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt caccagggtg
     1561 tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg
     1621 gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg
     1681 tggtcggggt agcggctgaa gcactgcacg ccgtaggtca gggtggtcac gagggtgggc
     1741 cagggcacgg gcagcttgcc ggtggtgcag atgaacttca gggtcagctt gccgtaggtg
     1801 gcatcgccct cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc gccgtccagc
     1861 tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac catggtggcg
     1921 accggtggat cccgggcccg cggtaccgcg aagacgtctt taccgaccga atctgaaggt
     1981 ttattgagtt caaccctcac acctttttca cctgttagat attttacttt agctcgtgct
     2041 ctctcatctg catcaaaata ccaaacagtt attgcgtacc ttgtagcata tgctggttgt
     2101 acttcatgag ggttgcgacg gtcagaccag aaaaacagca gtctatcaaa tttgggttca
     2161 atgtcagcaa actgggcttt gccttctgga aaaattcgaa gtatacctcc acttaccttg
     2221 gcatcccagt ctttattaag ataatatata catgtcacac atcttccatc tccatttgga
     2281 ttatcaacat tacgtacata acccgttcca ttgcccggat aacaagcgac catggctttc
     2341 gtccggccat tgattttgta gctgcccagc ttcccgttac agtggcgtat caggtcgtcc
     2401 atgctgctca tgagcagccc aatggtttcg cagccgggct ccttgccctc gatccaggtg
     2461 atcttatcgc ctcggatgtc cttggacgag tcactcttct ggctgaccag ctgcccgtcc
     2521 gtgaacttcc cggtgtcgtg cagggcgcgc acctcgtcgc cgatctgctg tccggtctcc
     2581 ttgccgagga agtcgtccac cacacagatg ccgtgcttgt tcatgcacgg cacgatgtac
     2641 tcgagcgcca gcttcagcgc cggcaggggc ttcgtctgcc cgttgggccg caggccgccg
     2701 ccggggctca gcgcatcccc gggcgtgttg cttggggggt acaggttcgc cttctcctgg
     2761 aacagcgatg agcgggccgg cggctcctcc ttgccgggct cggcttcggc agccaccgcc
     2821 gagccctggc cgccggcggc cgcacgacac ggcgacgcgg ccgccgctgg gtcggccggg
     2881 ggcttggcct ttacttttcc cttggccgcg tccccggagg cgttgtcccg gcgcgccgct
     2941 gccttcctgg gctcccgggc cccggccctg ggcggcggca ctgcagccgg cggcgcgggg
     3001 ccggaatgct ggtgtgggcc cactccgtgg ccgagggcgc cctcgctgcc ctggcacacg
     3061 agcttgtgct tcttccagtc ctgacgctgg tgctccttgc agcagtagaa ggagctgcgg
     3121 cagcggctgc agcgcagcag gttctccatc ttcccgcaca gctcgcagta ctgccggtct
     3181 cgctcgctcg ggctcggccc gccgggcccg ccgctgtcat tggccatggc ggcaagcttg
     3241 agctcgagat ctgagtccgg tagcgctagc ggctgagaga atagggcctg tgcggcgaat
     3301 ggcaacctgg gctcaggcgc gcgggcctgg ggagcgcaag accggccccc tcggccgccg
     3361 ccgccgcctc agcgtcccgg gcggcccggc ccaggctgtg gaggagcgca gggcatacgg
     3421 gcgccactcg ctctgcgcgc tctgcacgta caccacggcc ccgcggcgac ccgggcgggc
     3481 ggcgcgcgag ggcggagggg gcggagggag ggccgggagg gccgggggga gggccgaggg
     3541 gcgggggctg cctgcgctcc tccgcactcg gcggggccgg gcctcggctt ctgcctcgga
     3601 ggtgggtcgc ccgctttccc tccgcccggg accggactgg gacgccgaga ggccggtgcg
     3661 ctacctcagc ccgcagcggc ctcctcgggg acgccgaggc gcaggcccgc gagcggcgga
     3721 aaggcgtggg gcagcgcggc agcgcctaca cctgctggag gagggcgcca gccgcaagcg
     3781 ggtagcggag cgaaggcggc gtaatcatgg ctgctcttcc tccaggccgg cgcggggtgt
     3841 gcgaggctcg ctccagtcaa ggttgcaagg tggctattgc gatcctcatt ttgcagagga
     3901 ggaaacggaa attcagagaa gtaaagtaac ttgcccaagg tcacgcaaaa ctgggactga
     3961 atccaggaat caggctccca gattccaagg ctgttttttc cattacacca tacagcctct
     4021 atttcaagaa gccctgaaat ggttaagaac aaaataagta ttactatggt gttaatccaa
     4081 tctcttgcgc cccgtttcga cctagtttaa aggtaatgaa atttgagggt tttaaaaaaa
     4141 atcaaaaact catttttaaa acctttgaaa tcccgccgag ttcaaaaccc catgaagaaa
     4201 ttttggatta ataactaatg catggcggta atacggttat ccacagaatc aggggataac
     4261 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4321 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4381 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4441 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4501 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4561 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4621 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4681 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4741 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4801 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4861 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4921 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4981 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5041 tgaagtttta aatcaatcta aagtatatat gagtaacctg aggctatggc agggcctgcc
     5101 gccccgacgt tggctgcgag ccctgggcct tcacccgaac ttggggggtg gggtggggaa
     5161 aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg gtggggtatc gacagagtgc
     5221 cagccctggg accgaacccc gcgtttatga acaaacgacc caacaccgtg cgttttattc
     5281 tgtcttttta ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca
     5341 gttagcctcc ccctagggtg ggcgaagaac tccagcatga gatccccgcg ctggaggatc
     5401 atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa ggcggcggtg
     5461 gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc gaaccccaga
     5521 gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag
     5581 cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa
     5641 tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt
     5701 cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat
     5761 gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg
     5821 ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca
     5881 tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg
     5941 gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag
     6001 caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc
     6061 ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg
     6121 atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa
     6181 aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg
     6241 tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt
     6301 gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcgatcttt
//
