LOCUS       sequence-415465-        5122 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hypoxia responsive promoter driving destablized UnaG fluorescent 
            protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5122)
  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 5122)
  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-dUnaG
FEATURES             Location/Qualifiers
     source          1..5122
                     /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             914..943
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(979..996)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    985..1209
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1255..1683
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1342..1361)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1552..1573
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1692..1712)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1697..2026
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1877..2012
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1939..1958
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2052..2846
                     /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(2106..2125)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2716..2735
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3020..3141
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3057..3076)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3111..3130
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3174..3191)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3345..3933)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3425..3444)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4104..4895,4896..4964))
                     /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     4727..4746
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4965..5069)
                     /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 gtcgacccgc ggggatccac
      901 gcgtgcggcc gcagaacaaa aactcatctc agaagaggat ctgaatgggg ccgcatagtc
      961 tagaagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     1021 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     1081 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     1141 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatggcccgg
     1201 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     1261 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     1321 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     1381 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     1441 tacggcacct ctcccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     1501 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     1561 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     1621 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     1681 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     1741 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     1801 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     1861 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccct
     1921 aggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     1981 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tccccccccg
     2041 ggaggtccac aatggttgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     2101 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     2161 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     2221 tgaatgaact ccaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     2281 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     2341 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     2401 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     2461 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     2521 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     2581 gtatgcccga cggcgaggat ctcgtcgtga ctcatggcga tgcctgcttg ccgaatatca
     2641 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     2701 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     2761 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     2821 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     2881 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     2941 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     3001 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     3061 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     3121 caaactcatc aatgtatctt atcatgtctg tataccgtcg atctttccgc ttcctcgctc
     3181 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3241 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3301 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3361 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3421 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3481 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3541 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtatg
     3601 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3661 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3721 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3781 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3841 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3901 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3961 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4021 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4081 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4141 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4201 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4261 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4321 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4381 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4441 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4501 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     4561 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4621 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4681 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     4741 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4801 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgggcacc caactgatct
     4861 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4921 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4981 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5041 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     5101 agatcgacgg atcgggagat cg
//
