LOCUS       sequence-408663-        5836 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hypoxia responsive promoter driving UnaG and mOrange.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5836)
  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 5836)
  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-dUnOHR
FEATURES             Location/Qualifiers
     source          1..5836
                     /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             770..1477
                     /codon_start=1
                     /product="photostable monomeric orange derivative of DsRed 
                     fluorescent protein (Shaner et al., 2008)"
                     /label=mOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     CDS             1628..1657
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1693..1710)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1699..1923
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1969..2397
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2056..2075)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2266..2287
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2406..2426)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2411..2740
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2591..2726
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2653..2672
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2766..3560
                     /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(2820..2839)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3430..3449
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3734..3855
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3771..3790)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3825..3844
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3888..3905)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4059..4647)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4139..4158)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4818..5609,5610..5678))
                     /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     5441..5460
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5679..5783)
                     /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 tgtcgacccg cggggatcca tggtgagcaa
      781 gggcgaggag aataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcgcatgga
      841 gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga
      901 gggctttcag accgctaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga
      961 catcctgtcc cctcatttca cctacggctc caaggcctac gtgaagcacc ccgccgacat
     1021 ccccgactac ttcaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaacta
     1081 cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat
     1141 ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtga tgcagaagaa
     1201 gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggtg ccctgaaggg
     1261 caagatcaag atgaggctga agctgaagga cggcggccac tacacctccg aggtcaagac
     1321 cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacatcgtcg acatcaagtt
     1381 ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg
     1441 ccgccactcc accggcggca tggacgagct gtacaagacg cgtaagaagc ttagccacgg
     1501 cttcccgccg gaggtggagg agcaggatga tggcacgctg cccatgtctt gtgcccagga
     1561 gagcgggatg gaccgtcacc ctgcagcctg tgcttctgct aggatcaatg tgatttaggc
     1621 ggccgcagaa caaaaactca tctcagaaga ggatctgaat ggggccgcat agtctagaag
     1681 ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     1741 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     1801 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     1861 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatggc ccgggctcta
     1921 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     1981 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2041 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2101 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2161 acctctcccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     2221 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2281 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     2341 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     2401 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     2461 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     2521 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     2581 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc ccctaggctg
     2641 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     2701 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctccccc cccgggaggt
     2761 ccacaatggt tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     2821 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     2881 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     2941 aactccagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     3001 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     3061 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     3121 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     3181 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     3241 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgtatgc
     3301 ccgacggcga ggatctcgtc gtgactcatg gcgatgcctg cttgccgaat atcatggtgg
     3361 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     3421 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     3481 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     3541 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     3601 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     3661 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     3721 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     3781 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     3841 catcaatgta tcttatcatg tctgtatacc gtcgatcttt ccgcttcctc gctcactgac
     3901 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3961 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4021 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4081 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4141 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4201 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4261 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tatgcacgaa
     4321 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4381 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4441 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4501 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4561 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4621 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4681 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4741 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4801 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4861 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4921 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4981 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5041 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5101 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5161 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5221 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5281 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5341 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5401 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5461 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5521 ttaccgctgt tgagatccag ttcgatgtaa cccactcggg cacccaactg atcttcagca
     5581 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5641 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5701 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5761 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcagatcg
     5821 acggatcggg agatcg
//
