LOCUS       sequence-415471-        5941 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hypoxia responsive promoter driving UnaG fluorescent protein 
            destablized using ODDD fragment from HIF1a and a PEST sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5941)
  AUTHORS   Erapaneedi R, Belousov VV, Schafers M, Kiefer F
  TITLE     A novel family of fluorescent hypoxia sensors reveal strong 
            heterogeneity in tumor hypoxia at the cellular level.
  JOURNAL   EMBO J. 2016 Jan 4;35(1):102-13. doi: 10.15252/embj.201592775. Epub 
            2015 Nov 23.
  PUBMED    26598532
REFERENCE   2  (bases 1 to 5941)
  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: 
            "10.15252/embj.201592775"; journalName: "EMBO J"; date: "2016-01-4- 
            4"; volume: "35"; issue: "1"; pages: "102-13"
COMMENT     Sequence Label: HRE-doUnaG
FEATURES             Location/Qualifiers
     source          1..5941
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        229..267
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     264..288
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      323..332
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             329..745
                     /codon_start=1
                     /product="bilirubin-binding green fluorescent protein from 
                     Anguilla japonica (Kumagai et al., 2013)"
                     /label=UnaG
                     /translation="MVEKFVGTWKIADSHNFGEYLKAIGAPKELSDGGDATTPTLYISQ
                     KDGDKMTVKIENGPPTFLDTQVKFKLGEEFDEFPSDRRKGVKSVVNLVGEKLVYVQKWD
                     GKETTYVREIKDGKLVVTLTMGDVVAVRSYRRATE"
     CDS             1733..1762
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1798..1815)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1804..2028
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2074..2502
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2161..2180)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2371..2392
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2511..2531)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2516..2845
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2696..2831
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2758..2777
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2871..3665
                     /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="MVEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2925..2944)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3535..3554
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3839..3960
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3876..3895)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3930..3949
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3993..4010)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4164..4752)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4244..4263)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4923..5714,5715..5783))
                     /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     5546..5565
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5784..5888)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtaccgagct tttctcgagc cacagtgcat acgtgggctc caacaggtcc tcttgtcgag
       61 ccacagtgca tacgtgggct ccaacaggtc ctcttgtcga gccacagtgc atacgtgggc
      121 tccaacaggt cctcttgtcg agccacagtg catacgtggg ctccaacagg tcctcttgtc
      181 gagccacagt gcatacgtgg gctccaacag gtcctcttgt cgagatctgg taggcgtgta
      241 cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcac tagaagcttg
      301 gcattccggt actgttggta aagccaccat ggtcgagaaa tttgttggca cctggaagat
      361 cgcagacagc cataattttg gtgaatacct gaaagctatc ggagccccaa aggaattaag
      421 cgatggtggg gatgccacga cgccgacatt gtacatctcc cagaaggacg gagacaaaat
      481 gacagtgaaa atagagaatg gacctcctac gttccttgac actcaagtaa agttcaaatt
      541 aggggaggag ttcgacgaat ttccttctga tcgaagaaaa ggcgtaaaat ctgtcgtgaa
      601 cttggtggga gagaagctgg tgtacgtaca aaagtgggac ggcaaggaga cgacgtatgt
      661 ccgagagata aaggacggta aactggtcgt gacacttacg atgggagacg tcgtggctgt
      721 gcgcagctac cggagggcga cggaaaccgg taagaagctt agccacggct tcccgccgga
      781 ggtggaggag caggatgatg gcacgctgcc catgtcttgt gcccaggaga gcgggatgga
      841 ccgtcaccct gcagcctgtg cttctgctag gatcaatgtg gtcgacggag tgaattacgt
      901 tgtgagtggt attattcagc acgacttgat tttctccctt caacaaacag aatgtgtcct
      961 taaaccggtt gaatcttcag atatgaaaat gactcagcta ttcaccaaag ttgaatcaga
     1021 agatacaagt agcctctttg acaaacttaa gaaggaacct gatgctttaa ctttgctggc
     1081 cccagccgct ggagacacaa tcatatcttt agattttggc agcaacgaca cagaaactga
     1141 tgaccagcaa cttgaggaag taccattata taatgatgta atgctcccct cacccaacga
     1201 aaaattacag aatataaatt tggcaatgtc tccattaccc accgctgaaa cgccaaagcc
     1261 acttcgaagt agtgctgacc ctgcactcaa tcaagaagtt gcattaaaat tagaaccaaa
     1321 tccagagtca ctggaacttt cttttaccat gccccagatt caggatcaga cacctagtcc
     1381 ttccgatgga agcactagac aaagttcacc tgagcctaat agtcccagtg aatattgttt
     1441 ttatgtggat agtgatatgg tcaatgaatt caagttggaa ttggtagaaa aactttttgc
     1501 tgaagacaca gaagcaaaga acccattttc tactcaggac acagatttag acttggagat
     1561 gttagctccc tatatcccaa tggatgatga cttccagtta cgttccttcg atcagttgtc
     1621 accattagaa agcagttccg caagccctga aagcgcaagt cctcaaagca cagttacagt
     1681 attccagcag actcaaatac aaggaccgcg gggatccacg cgtgcggccg cagaacaaaa
     1741 actcatctca gaagaggatc tgaatggggc cgcatagtct agaagctcgc tgatcagcct
     1801 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     1861 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     1921 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     1981 attgggaaga caatagcagg catgctgggg atggcccggg ctctatggct tctgaggcgg
     2041 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     2101 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     2161 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     2221 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc tcccccaaaa
     2281 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     2341 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     2401 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     2461 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     2521 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2581 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2641 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     2701 gcccctaact ccgcccagtt ccgcccattc tccgccccta ggctgactaa ttttttttat
     2761 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     2821 ttttggaggc ctaggctttt gcaaaaagct ccccccccgg gaggtccaca atggttgaac
     2881 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     2941 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3001 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactc caggacgagg
     3061 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3121 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3181 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3241 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3301 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3361 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg tatgcccgac ggcgaggatc
     3421 tcgtcgtgac tcatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     3481 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     3541 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     3601 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     3661 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     3721 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     3781 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     3841 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3901 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3961 tcatgtctgt ataccgtcga tctttccgct tcctcgctca ctgactcgct gcgctcggtc
     4021 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4081 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4141 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4201 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4261 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4321 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4381 agttcggtgt aggtcgttcg ctccaagctg ggctgtatgc acgaaccccc cgttcagccc
     4441 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4501 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4561 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4621 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4681 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4741 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4801 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4861 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4921 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4981 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5041 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5101 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5161 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5221 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5281 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5341 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5401 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5461 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5521 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5581 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5641 tccagttcga tgtaacccac tcgggcaccc aactgatctt cagcatcttt tactttcacc
     5701 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5761 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5821 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5881 gttccgcgca catttccccg aaaagtgcca cctgacgtca gatcgacgga tcgggagatc
     5941 g
//
