LOCUS       sequence-325110-        5333 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector for: Peredox NADH/NAD+ sensor (cytosolic).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5333)
  AUTHORS   Werley CA, Boccardo S, Rigamonti A, Hansson EM, Cohen AE
  TITLE     Multiplexed Optical Sensors in Arrayed Islands of Cells for 
            multimodal recordings of cellular physiology.
  JOURNAL   Nat Commun. 2020 Aug 4;11(1):3881. doi: 10.1038/s41467-020-17607-5.
  PUBMED    32753572
REFERENCE   2  (bases 1 to 5333)
  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"; journalName: "Nat 
            Commun."; date: "2020-08-4"; volume: "11(1):3881. doi"; pages: " 
            10.1038/s41467-020-17607-5"
COMMENT     Sequence Label: pMOS023E: Entry vector for: Peredox NADH/NAD+ ...
FEATURES             Location/Qualifiers
     source          1..5333
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(688..1299,1300..1320,1321..2055,2056..2058,2059..2688,
                     2689..2733)
                     /codon_start=1
                     /product="genetically encoded fluorescent biosensor of the 
                     cytosolic NADH-NAD+ redox state (Hung et al., 2011)"
                     /label=Peredox
                     /note="consists of two pH-insensitive mutant copies of the 
                     Thermus aquaticus redox-sensing repressor Rex (T-Rex) 
                     separated by circularly permuted monomeric T-Sapphire 
                     (cpmTS), a pH-resistant GFP variant"
                     /translation="KVPEAAISRLITYLRILEELEAQGVHRTASEQLGELAQVTAFQVD
                     KDLSYFGSYGTDGVGYTVPVLKRELRHILGLNRKWGLCIVGMGRLGSALADWPGFGESF
                     ELRGFFDVDPGMVGRPVRGGVIEHVDLLPQRVPGRIEIALLTVPREAAQKAADLLVAAG
                     IKGILNFAPVVLEVPKEVAVENVDILAGLTRLSFAILNPTWSAAGGHGFTAHNVYIMAD
                     KQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSIQSKLSKDPNEK
                     RDHMVLLEFVTAAGITHGMDELYKGGTGGSMVSKGEELFTGVVPILVELDGDVNGHKFS
                     VSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFSYGVMVFARYPDHMKQHDFFKSA
                     MPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNTK
                     VPEAAISRLITYLRILEELEAQGVHRTASEQLGELAQVTAFQVDEDLSYFGSYGTDGVG
                     YTVPVLKRELRHILGLNRKWGLCIVGMGRLGSALADWPGFGESFELRGFFDVDPGMVGR
                     PVRGGVIEHVDLLPQRVPGRIEIALLTVPREAAQKAADLLVAAGIKGILNFAPVVLEVP
                     KEVAVENVDFLAGLTRLSFAILNPKWREEMMGSGTGGNASDGGGSGG"
     CDS             2734..3444
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2879..2897)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3147..3166
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(3458..3557)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3574..3593)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3575..3593)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3598..3614)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3598..3614)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3716..3735)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3727..4536
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3794..3813)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4683..5271
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5172..5191
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctgc ccaccatggt tgctagcaaa gttcctgaag cagccatttc cagactgatt
      721 acttacctga gaattctgga agagctggaa gcccaaggtg tacaccgcac cgcctccgag
      781 caactgggcg agctggccca ggtcaccgcc ttccaggttg ataaggacct gtcctacttc
      841 ggctcctacg gcactgacgg cgttggctac actgtaccgg tcctcaagag agaactgcgc
      901 catatcctcg gtctgaaccg caaatggggc ctgtgcatcg tggggatggg ccgcctggga
      961 tccgctcttg ctgattggcc tggttttggc gagagcttcg agctgcgcgg tttcttcgat
     1021 gttgacccag gtatggttgg tcggccggtt cgcggtggtg ttatcgaaca cgttgatctg
     1081 ctgccccagc gcgtacctgg tcgcatcgaa atcgctctgc tgacggttcc acgcgaggca
     1141 gcacagaagg ctgccgacct gctggttgcc gctggcatca agggcatcct gaacttcgct
     1201 ccagttgtac tggaggttcc caaggaagtg gctgttgaga acgttgacat cctggccggt
     1261 ctgacgcgtc tgagctttgc cattctgaac cccacgtgga gcgcagcagg tgggcatggt
     1321 ttcactgctc acaacgtgta tatcatggct gacaaacaga agaacggtat caaggccaac
     1381 ttcaagatcc gccacaacat cgaagacggc ggcgttcagc tggctgatca ctaccagcag
     1441 aacaccccaa tcggcgacgg ccctgttctg ctgcctgaca accactactt aagcatccag
     1501 tctaaactgt ccaaagaccc aaacgagaag cgcgaccaca tggttctgct ggaatttgtt
     1561 accgccgctg gtatcaccca cggcatggac gaactgtaca agggcggcac cgggggatct
     1621 atggtttcca aaggcgagga actgttcact ggtgttgttc ctatcctggt tgaactggac
     1681 ggcgacgtta acggccacaa attcagcgtt agcggggaag gcgaaggaga cgctacctac
     1741 ggcaagctga ctctgaagtt catctgcacc actggcaagc tgcctgttcc ttggcctacc
     1801 ctggttacta ccttctccta tggtgttatg gttttcgcca gatatcctga ccacatgaag
     1861 cagcacgact tcttcaagag cgccatgcct gagggctacg ttcaggaacg caccatcttt
     1921 ttcaaggacg acggcaacta taagacacgc gctgaggtta agttcgaggg cgacactctg
     1981 gttaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctgggccat
     2041 aagcttgaat ataacaccaa ggtacctgaa gccgccatca gccgcctcat cacttacctc
     2101 cgcatcctgg aggagctcga agcccaaggc gttcaccgca ctgcaagcga gcagctgggc
     2161 gaactggcac aggttacagc attccaggtt gacgaagatc tgtcctattt cggttcctac
     2221 ggcaccgacg gtgttggcta cacagtccct gtcctgaagc gcgagctgcg ccacatcctg
     2281 ggcctgaacc gcaagtgggg cttatgcatc gttggcatgg gccgcctcgg tagtgcactg
     2341 gcagactggc cgggctttgg tgaatctttc gaactgcgcg gcttcttcga cgttgatcca
     2401 ggcatggttg ggcgccctgt tcgcggtggc gttatcgagc atgttgacct cctgccacag
     2461 cgcgtcccgg gtcgcatcga gatcgccctg ctgaccgttc ctcgcgaagc tgcacagaag
     2521 gcagccgact tactagtcgc cgctggtatc aagggaattc tgaacttcgc ccctgttgtt
     2581 ctggaggttc ctaaggaagt tgctgttgaa aacgttgatt tcctggccgg cctgacccgc
     2641 ctgagcttcg caatcctgaa tcctaagtgg cgcgaagaaa tgatgggttc cggaactggg
     2701 ggcaacgcct ctgacggtgg tgggtctggt ggtatggtga gcaagggcga ggaggataac
     2761 atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc
     2821 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     2881 aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag
     2941 ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag
     3001 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
     3061 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg
     3121 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     3181 gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg
     3241 ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag
     3301 aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac
     3361 aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc
     3421 ggcatggacg aactgtacaa gtgagtcgag aagcttaacc cagctttctt gtacaaagtt
     3481 ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa
     3541 aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta ttacatggtc
     3601 atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa
     3661 gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
     3721 ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat
     3781 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     3841 tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     3901 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
     3961 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     4021 atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     4081 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     4141 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
     4201 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     4261 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     4321 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     4381 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     4441 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     4501 ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa
     4561 cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt
     4621 aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4681 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4741 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4801 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4861 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4921 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4981 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5041 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5101 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5161 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5221 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5281 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gtt
//
