LOCUS       sequence-380466-        7708 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mTurq2-Apollo-NADP+ sensor localized to the peroxisome.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7708)
  AUTHORS   Chang HH, Bennett AM, Cameron WD, Floro E, Au A, McFaul CM, Yip CM, 
            Rocheleau JV
  TITLE     Targeting Apollo-NADP(+) to Image NADPH Generation in Pancreatic 
            Beta-Cell Organelles.
  JOURNAL   ACS Sens. 2022 Oct 21. doi: 10.1021/acssensors.2c01174.
  PUBMED    36269889
REFERENCE   2  (bases 1 to 7708)
  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: "ACS Sens. 
            2022 Oct 21. doi: 10.1021/acssensors.2c01174."
COMMENT     Sequence Label: Perox-mTurq2-Apollo-NADP+
FEATURES             Location/Qualifiers
     source          1..7708
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(55..74)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        246..625
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        626..829
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     780..800
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     874..893
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        874..892
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      969..978
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2529..2538
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2535..2537,2538..2540,2541..3251)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2580..2601)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2841..2860)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3188..3209
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3310..3327)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3316..3540
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3586..4014
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3673..3692)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3883..3904
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4023..4043)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4028..4357
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4208..4343
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4270..4289
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4424..5218
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4478..4497)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5088..5107
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5392..5513
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5429..5448)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5483..5502
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5562..5578)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5562..5578)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5575..5597)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5586..5602
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5610..5616,5617..5634,5635..5640))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5655..5676
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5793..5810)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5964..6552)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6044..6063)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6723..7514,7515..7583))
                     /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     7346..7365
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7584..7688)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
       61 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
      121 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
      181 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
      241 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
      301 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      361 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      421 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      481 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      541 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      601 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      661 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      721 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      781 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac
      841 tagagaaccc actgcttact ggcttatcga aattaatacg actcactata gggagaccca
      901 agctggctag cgtttaaact taagcttggt accgagctcg gatccactag tccagtgtgg
      961 tggaattcgc caccatggca gagcaggtgg ccctgagccg gacccaggtg tgcgggatcc
     1021 tgcgggaaga gcttttccag ggcgatgcct tccatcagtc ggatacacac atattcatca
     1081 tcatgggtgc agcgggtgac ctggccaaga agaagatcta ccccaccatc tggtggctgt
     1141 tccgggatgg ccttctgccc gaaaacacct tcatcgtggg ctatgctcag tcccgcctca
     1201 cagtggctga catccgcaaa cagagtgagc ccttcttcaa ggccacccca gaggagaagc
     1261 tcaagctgga ggacttcttt gcccgcaact cctatgtggc tggccagtac gatgatgcag
     1321 cctcctacca gcgcctcaac agccacatga atgccctcca cctggggtca caggccaacc
     1381 gcctcttcta cctggccttg cccccgaccg tctacgaggc cgtcaccaag aacattcacg
     1441 agtcctgcat gagccagata ggctggaacc gcatcatcgt ggagaagccc ttcgggaggg
     1501 acctgcagag ctctgaccgg ctgtccaacc acatctcctc cctgttccgt gaggaccaga
     1561 tctaccgcat cgacaactac ctgggcacgg agatggtgca gaacctcatg gtgctgagat
     1621 ttgccaacag gatcttcggc cccatctgga accgggacaa catcgcctgc gttatcctca
     1681 ccttcaagga gccctttggc actgagggtc gcgggggcta tttcgatgaa tttgggatca
     1741 tccgggacgt gatgcagaac cacctactgc agatgctgtg tctggtggcc atggagaagc
     1801 ccgcctccac caactcagat gacgtccgtg atgagaaggt caaggtgttg aaatgcatct
     1861 cagaggtgca ggccaacaat gtggtcctgg gccagtacgt ggggaacccc gatggagagg
     1921 gcgaggccac caaagggtac ctggacgacc ccacggtgcc ccgcgggtcc accaccgcca
     1981 cttttgcagc cgtcgtcctc tatgtggaga atgagaggtg ggatggggtg cccttcatcc
     2041 tgcgctgcgg caaggccctg aacgagcgca aggccgaggt gaggctgcag ttccatgatg
     2101 tggccggcga catcttccac cagcagtgca agcgcaacga gctggtgatc cgcgtgcagc
     2161 ccaacgaggc cgtgtacacc aagatgatga ccaagaagcc gggcatgttc ttcaaccccg
     2221 aggagtcgga gctggacctg acctacggca acagatacaa gaacgtgaag ctccctgacg
     2281 cctatgagcg cctcatcctg gacgtcttct gcgggagcca gatgcacttc gtgcgcagcg
     2341 acgagctccg tgaggcctgg cgtattttca ccccactgct gcaccagatt gagctggaga
     2401 agcccaagcc catcccctat atttatggca gccgaggccc cacggaggca gacgagctga
     2461 tgaagagagt gggtttccag tatgagggca cctacaagtg ggtgaacccc cacaagctca
     2521 gaccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     2581 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     2641 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     2701 tgccctggcc caccctcgtg accaccctgt cctggggcgt gcagtgcttc gcccgctacc
     2761 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     2821 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     2881 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     2941 acatcctggg gcacaagctg gagtacaact actttagcga caacgtctat atcaccgccg
     3001 acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggcg
     3061 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     3121 tgcccgacaa ccactacctg agcacccagt ccaagctgag caaagacccc aacgagaagc
     3181 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     3241 agctgtacaa gcgccgccgc agcaagttat aactcgagtc tagagggccc gtttaaaccc
     3301 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     3361 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     3421 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     3481 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     3541 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     3601 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3661 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3721 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     3781 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     3841 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     3901 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     3961 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     4021 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4081 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4141 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4201 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4261 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     4321 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     4381 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     4441 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4501 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4561 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     4621 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4681 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4741 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4801 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4861 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     4921 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     4981 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     5041 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     5101 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     5161 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     5221 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     5281 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     5341 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     5401 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     5461 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     5521 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     5581 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     5641 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     5701 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     5761 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5821 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5881 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5941 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6001 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6061 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6121 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6181 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6241 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6301 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6361 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     6421 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6481 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6541 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6601 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6661 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6721 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6781 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6841 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6901 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6961 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7021 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7081 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7141 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7201 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7261 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7321 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7381 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7441 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7501 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7561 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7621 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7681 gttccgcgca catttccccg aaaagtgc
//
