LOCUS       sequence-415470-        5812 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hypoxia responsive promoter driving UnaG fluorescent protein 
            destablized using ODDD fragment from HIF1a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5812)
  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 5812)
  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-oUnaG
FEATURES             Location/Qualifiers
     source          1..5812
                     /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             1604..1633
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1669..1686)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1675..1899
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1945..2373
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2032..2051)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2242..2263
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2382..2402)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2387..2716
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2567..2702
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2629..2648
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2742..3536
                     /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(2796..2815)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3406..3425
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3710..3831
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3747..3766)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3801..3820
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3864..3881)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4035..4623)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4115..4134)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4794..5585,5586..5654))
                     /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     5417..5436
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5655..5759)
                     /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 tgtcgacgga gtgaattacg ttgtgagtgg
      781 tattattcag cacgacttga ttttctccct tcaacaaaca gaatgtgtcc ttaaaccggt
      841 tgaatcttca gatatgaaaa tgactcagct attcaccaaa gttgaatcag aagatacaag
      901 tagcctcttt gacaaactta agaaggaacc tgatgcttta actttgctgg ccccagccgc
      961 tggagacaca atcatatctt tagattttgg cagcaacgac acagaaactg atgaccagca
     1021 acttgaggaa gtaccattat ataatgatgt aatgctcccc tcacccaacg aaaaattaca
     1081 gaatataaat ttggcaatgt ctccattacc caccgctgaa acgccaaagc cacttcgaag
     1141 tagtgctgac cctgcactca atcaagaagt tgcattaaaa ttagaaccaa atccagagtc
     1201 actggaactt tcttttacca tgccccagat tcaggatcag acacctagtc cttccgatgg
     1261 aagcactaga caaagttcac ctgagcctaa tagtcccagt gaatattgtt tttatgtgga
     1321 tagtgatatg gtcaatgaat tcaagttgga attggtagaa aaactttttg ctgaagacac
     1381 agaagcaaag aacccatttt ctactcagga cacagattta gacttggaga tgttagctcc
     1441 ctatatccca atggatgatg acttccagtt acgttccttc gatcagttgt caccattaga
     1501 aagcagttcc gcaagccctg aaagcgcaag tcctcaaagc acagttacag tattccagca
     1561 gactcaaata caaggaccgc ggggatccac gcgtgcggcc gcagaacaaa aactcatctc
     1621 agaagaggat ctgaatgggg ccgcatagtc tagaagctcg ctgatcagcc tcgactgtgc
     1681 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     1741 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     1801 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     1861 acaatagcag gcatgctggg gatggcccgg gctctatggc ttctgaggcg gaaagaacca
     1921 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     1981 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2041 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2101 ggctcccttt agggttccga tttagtgctt tacggcacct ctcccccaaa aaacttgatt
     2161 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2221 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2281 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2341 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     2401 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2461 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2521 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     2581 tccgcccagt tccgcccatt ctccgcccct aggctgacta atttttttta tttatgcaga
     2641 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     2701 cctaggcttt tgcaaaaagc tccccccccg ggaggtccac aatggttgaa caagatggat
     2761 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     2821 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     2881 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact ccaggacgag gcagcgcggc
     2941 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     3001 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     3061 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     3121 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     3181 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     3241 cagccgaact gttcgccagg ctcaaggcgc gtatgcccga cggcgaggat ctcgtcgtga
     3301 ctcatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     3361 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     3421 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     3481 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     3541 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     3601 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     3661 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     3721 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3781 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     3841 tataccgtcg atctttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3901 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3961 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4021 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4081 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4141 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4201 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4261 taggtcgttc gctccaagct gggctgtatg cacgaacccc ccgttcagcc cgaccgctgc
     4321 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4381 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4441 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4501 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4561 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4621 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4681 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4741 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4801 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4861 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4921 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4981 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5041 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5101 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5161 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     5221 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5281 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5341 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5401 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5461 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5521 atgtaaccca ctcgggcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5581 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5641 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5701 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5761 acatttcccc gaaaagtgcc acctgacgtc agatcgacgg atcgggagat cg
//
