LOCUS       sequence-491315-        7768 bp    DNA     circular SYN 31-OCT-2025
DEFINITION  Polyubiquitin promoter driving expression of glutathione biosensor 
            GRX1-roGFP2. Flanked by ITRs compatible with minos-mediated 
            transgenesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7768)
  TITLE     Redox biosensor expression plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7768)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 31, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: LvPolyubiquitin-GRX1-roGFP2-minos-lox-ITR
FEATURES             Location/Qualifiers
     source          1..7768
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     742..764
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     761..777
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     761..777
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    complement(1174..1207)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             join(5102..5104,5105..5818)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FISTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNCHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTCSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5144..5165)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5405..5424)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5752..5773
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5847..5981
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5871..5890)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5925..5944
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(6005..6038)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     primer_bind     complement(6433..6450)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6433..6449)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6442..6464)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        6563..6667
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6668..6736,6737..7528)
                     /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     complement(6886..6905)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    7628..7652
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttccagc
      661 gacatatgcc atacgccgaa ctgaactcca cctaatgtct tggagattgt cggatagcgc
      721 ggaataggat tgtgccctct gagcggataa caatttcaca caggaaacag ctatgaccat
      781 gacacatgca tccaccatcg cagacttatc atcactgagc ctccacctag cctcattctc
      841 tatgctagct taagatattt ttcgttttct acgagcccca accactatta attcgaacag
      901 catgtttttt ttgcagtgcg caatgtttaa cacactatat tatcaatact actaaagata
      961 acacatacca atgcatttcg tctcaaagag aattttattc tcttcacgac gaaaaaaaaa
     1021 gttttgctct atttccaaca acaacaaaaa tatgagtaat ttattcaaac ggtttgctta
     1081 agagataaga aaaaagtgac cactattaat tcgaacgcgg cgtaagctac cttaatctca
     1141 agaagagcaa aacaaaagca actaatgtaa cggataactt cgtatagcat acattatacg
     1201 aagttataac catatgtgac gtctacggat ccgcatgcac tagtcctcac aacatttctt
     1261 cccttttttg cgggagttgg ggcaaataat cctgacaatg tccaatgatt attttaaaaa
     1321 gtgctatgta caatcgcttt gtatttgtat aattgttata aaaaatgcat tgtatgtatt
     1381 tatatttaca tacataatta ttctacaaag tgtattgtac aatcacttta tatcaacttt
     1441 agatgtatcg tgttattagt gtcaaatctg actgcgtgca tactgattcg atgccaaaag
     1501 tggattaatg cctccttgtg ccccgcgtgt ctcttggcaa ctagagcatt attgtagaat
     1561 attggggcac ctaaaataca ctgacaagca atcatccaaa aatatctaac aaaatactaa
     1621 acaataacag gacggtcatt tttctgatat ctaaaacaag tttttgttga agggacattg
     1681 tgcaagactt tcaaaaactt attaagaatg ttgaaccgtt tacttttttt attttaacac
     1741 gtgatactat agtatacata acgattgggc gaaaaaatac cgtgacagtc aatcaaacaa
     1801 tctaaattcc cactttgcag aatcactttt gtgattccct tgtatcttca acaagatata
     1861 gatagatgga tagcatggaa gtttttgttt tttacttaga aatggtaaaa cgatgttttt
     1921 tccccagggt cttttccccc tatgtaaatt atgtttcatt ctttcaataa aaaaaaacac
     1981 aaactgaaaa aaaacttata agcaaaaaaa aaggcgtaaa tattttttta tgaactctgt
     2041 taaacggaaa caacaacaac acagtcgcca atatatacct cccgcgacca gtgatcattc
     2101 cagaaaatgt tttgggccga aattatattg cataattccg aaattgcgag tcatcatcgt
     2161 ctgcaatatc tggcgcgggt taatatgacg tcatgtgcca tagatacaca atctatcttt
     2221 attacccaat agaaaaggat gaaatagtcc atgtgacaca gactgaccaa tcgcaagtca
     2281 acacaaagca ccatataaaa cacacgcaag cgtcgcagtg tcaataaatt gcatgagact
     2341 tccgcagagg ttgcactcga gaagaactcg tccatttttt tccagaaaaa ctctaatttc
     2401 aaagtaagta gacgaggttt tcacattatt ttaaggcagt tatttcaatg aagatgcagt
     2461 ttgtaaatgt ttgatttaaa aaaagcactt ttgaagaaga atgatgatga agaacataaa
     2521 ttgtcaacga ccacatcgtc gtcattattc gtgtgctttt tttttgttgt aatttatgtt
     2581 tggctttcca aatgaaggca atataggaac attaacagca tgaacaaagt ctcatattta
     2641 gtttgattga aagttcaaat tatagggcgt agaaatcatt gaaattgtag ggtggatacc
     2701 gtggattatt gcgtcgtgtg tgtgcatgat aagatcgtct gactcagacg tcgcgtcttt
     2761 tgttgtgtgg tgtgtgtggt atggatgatg atcaattgtg atgtttggtg gggattttca
     2821 tgaacgctga attaaaagac aatgagtgga aatggccatg ccaatttctg gtccatggat
     2881 tgtgtgaatg caatgacaat ttcattcaat ttttgtctgg gctgccatga tgaacatgat
     2941 gaagtctgaa ctgaagtgtt gtgattgaaa tgacaaagaa tttttgtcat gagaaagaat
     3001 gaggcatgta cggtacggta acttaaaacg ttgatagcta ggtgtagatt ttctagatac
     3061 gtttggcagt aggcctagat ctagaggcac ttggccaatc atgaatagtc tatgggagac
     3121 aatttcccat attgaagact tgctttgcaa aagaaacgac accaacaaag cgtaattttt
     3181 tccaccccaa attttttcgc gaaggtcaga acagaagccc atggcaaagg cctgttttgt
     3241 atgtgcatgg ccatggatga aaaaaatata aatacaaaag ttggtgaatt tttaaagatg
     3301 aaaaggggta atttttcatg aggtatctgc ttaggcccta tcacattttt atttgctgcc
     3361 aaggtaattc ttttgctgca tgtgtaaaaa aaggaaattg gtctcccaaa ctgggagtgg
     3421 agactgtctc tgaaattgga tacccaagtt ctttaatctt tacaaaattc tgatattgaa
     3481 aataatctct ctcatgggag atgttctccc atattggctg tgcaccagtt ggtcggtgtg
     3541 caaagtctag acaatacgta tccaggcact atcgcctatg aataatgcaa tagcagtgca
     3601 gttcacaacg gcctctctcc cgatcagcgc agatgttggg gaactaattt caacttttgt
     3661 cgtaatttat gtcaaataac tgacagtcgt aacttaagac cctaatcagc taatgttaat
     3721 ccttgacctg aattgcaggc atctgaaacg gctgtaatcc agcttcgatc ttaatgtcat
     3781 agttcaaatt ctcagtagat ttaactgagc gtggtgcgtg tttggccatt gccatagttt
     3841 ttatatggac tgaagtcaac gaccaaaaga tctgctcaag gcagcccaac tgcattagca
     3901 agattttggt tgctattcag tcaatatcag aactacggca atggacgagt gcggcgatgt
     3961 caacctacat gtaactgaga attcaaacta tgacattgag aacgaagcca aaatacagca
     4021 aatattgatg tggttgaaac ttgggtcaag gatcaaatgg acaatatttg gcaaaaaaga
     4081 taaattttga ggtgaaattg tttcctgaca ttgccatgtt ggttggtgtg aaactgcagc
     4141 gcaagacaat gaaggaagag actttataga gtaagctgcc ccgaatttga catgttttga
     4201 accatttttt gttcttaagt cttttcacat aatgaaaatt gcattaaaat gatcaatgtt
     4261 ggttgaatat atttagaatt gcaatgttga atttctccat tttgaatatt tcataaaaaa
     4321 tctgcactgt accaccatgt tttgctcccg ccacagattt cttaatttta gagcatttta
     4381 tggctacctc atttcttaaa tctgagcaaa gctaccgcat gttattgaaa tatggcacag
     4441 attcgcattt ttaaagggga ttttggtgtt caagtcgtga cttgcatgtg gtgggtgata
     4501 atgcctgtgt atgtcaatgc atgtggttcg tgttgtgtgg attgactgag ctgacttgtg
     4561 gaataagttt gccatgttga attccaggca tgctactctg aataaaggtc tttagaaaaa
     4621 cagattccaa taatgttttc atttttttgt caattttcta caggtcagtg cgatcgcatc
     4681 gatggctcaa gagtttgtga actgcaaaat ccagcctggg aaggtggttg tgttcatcaa
     4741 gcccacctgc ccgtactgca ggagggccca agagatcctc agtcaattgc ccatcaaaca
     4801 agggcttctg gaatttgtcg atatcacagc caccaaccac actaacgaga ttcaagatta
     4861 tttgcaacag ctcacgggag caagaacggt gcctcgagtc tttattggta aagattgtat
     4921 aggcggatgc agtgatctag tctctttgca acagagtggg gaactgctga cgcggctaaa
     4981 gcagattgga gctctgcaga ctagtggtgg ttcaggtggt ggtggttcag gtggtggtgg
     5041 ttcaggtgga ggaggatcag gaggaggagg atcaggagga ggaggatcag gaggagaatt
     5101 cgtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     5161 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     5221 caagctgacc ctgaagttca tctccaccac cggcaagctg cccgtgccct ggcccaccct
     5281 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     5341 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     5401 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     5461 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     5521 gctggagtac aactacaact gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     5581 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     5641 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     5701 cctgagcacc tgctccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     5761 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaact
     5821 agaactatag tgagtcgtat tacgtagatc cagacatgat aagatacatt gatgagtttg
     5881 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     5941 ttgctttatt tgtaaccatt ataagctgca ataaacaagt tggccgcctt cacgcgtggt
     6001 taccataact tcgtatagga tactttatac gaagttatcc gcgggggaaa ttaattaaat
     6061 tattgtttta agtatgatag taaatcacat tacgccgcgt tcgaattaat agtggtcact
     6121 tttttcttat ctcttaagca aaccgtttga ataaattact catatttttg ttgttgttgg
     6181 aaatagagca aaactttttt tttcgtcgtg aagagaataa aattctcttt gagacgaaat
     6241 gcattggtat gtgttatctt tagtagtatt gataatatag tgtgttaaac attgcgcact
     6301 gcaaaaaaac atgctgttcg aattaatagt ggttggggct cgtagaagct gctaacgtgc
     6361 ttggcataat ttatagaagc aggtcaggcg gaatggcact tccttattcc agatgcgtgc
     6421 ggattatgca tgactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgcttg
     6481 tggtggatca cagttattgg agacttcctt ggccgtacga tgggttttgt ttcaggtatc
     6541 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     6601 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     6661 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     6721 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     6781 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     6841 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     6901 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     6961 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     7021 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     7081 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     7141 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     7201 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     7261 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     7321 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     7381 tggctctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     7441 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     7501 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     7561 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc ttgaattgcc
     7621 attatcaaca agtttgtaca aaaaagcagg ctccgaattg gtatcacgag gacgatttga
     7681 agccccctac ccatttacgc tgacttgacg ggacggcggc tttgttgaat agagcgtcag
     7741 accccgtaga aaagatcaaa ggatcttc
//
