LOCUS       sequence-408666-        4993 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hypoxia responsive promoter driving UnaG fluorescent proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4993)
  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 4993)
  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-UnaG
FEATURES             Location/Qualifiers
     source          1..4993
                     /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             785..814
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(850..867)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    856..1080
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1126..1554
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1213..1232)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1423..1444
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1563..1583)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1568..1897
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1748..1883
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1810..1829
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1923..2717
                     /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(1977..1996)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2587..2606
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2891..3012
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2928..2947)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2982..3001
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3045..3062)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3216..3804)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3296..3315)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3975..4766,4767..4835))
                     /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     4598..4617
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4836..4940)
                     /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 cgcgtgcggc
      781 cgcagaacaa aaactcatct cagaagagga tctgaatggg gccgcatagt ctagaagctc
      841 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
      901 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
      961 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     1021 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatggcccg ggctctatgg
     1081 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     1141 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     1201 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     1261 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     1321 tctcccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     1381 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     1441 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     1501 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     1561 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     1621 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     1681 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     1741 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc taggctgact
     1801 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     1861 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccccccc gggaggtcca
     1921 caatggttga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     1981 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     2041 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     2101 tccaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     2161 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     2221 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     2281 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     2341 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     2401 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgtatgcccg
     2461 acggcgagga tctcgtcgtg actcatggcg atgcctgctt gccgaatatc atggtggaaa
     2521 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     2581 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     2641 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     2701 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     2761 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     2821 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     2881 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     2941 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3001 caatgtatct tatcatgtct gtataccgtc gatctttccg cttcctcgct cactgactcg
     3061 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3121 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3181 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3241 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3301 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3361 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3421 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtat gcacgaaccc
     3481 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3541 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3601 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3661 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3721 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3781 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3841 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3901 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3961 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4021 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4081 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4141 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4201 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4261 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4321 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4381 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4441 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4501 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4561 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     4621 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4681 ccgctgttga gatccagttc gatgtaaccc actcgggcac ccaactgatc ttcagcatct
     4741 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4801 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     4861 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4921 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cagatcgacg
     4981 gatcgggaga tcg
//
