LOCUS       sequence-380425-        7687 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mTurq2-Apollo-NADP+ sensor localized to the cytoplasm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7687)
  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 7687)
  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: Cyto-mTurq2-Apollo-NADP+
FEATURES             Location/Qualifiers
     source          1..7687
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(72..91)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        263..642
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        643..846
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     797..817
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     891..910
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        891..909
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      986..995
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2546..2555
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2552..2554,2555..2557,2558..3268)
                     /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(2597..2618)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2858..2877)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3205..3226
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3306..3323)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3312..3536
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3582..4010
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3669..3688)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3879..3900
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4019..4039)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4024..4353
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4204..4339
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4266..4285
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4420..5214
                     /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(4474..4493)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5084..5103
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5388..5509
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5425..5444)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5479..5498
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5558..5574)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5558..5574)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5571..5593)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5582..5598
                     /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(5606..5612,5613..5630,5631..5636))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5651..5672
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5789..5806)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5960..6548)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6040..6059)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6719..7510,7511..7579))
                     /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     7342..7361
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7580..7684)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
       61 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
      121 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
      181 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
      241 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
      301 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      361 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      421 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      481 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      541 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      601 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      661 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      721 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      781 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      841 agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact
      901 cactataggg agacccaagc tggctagcgt ttaaacttaa gcttggtacc gagctcggat
      961 ccactagtcc agtgtggtgg aattcgccac catggcagag caggtggccc tgagccggac
     1021 ccaggtgtgc gggatcctgc gggaagagct tttccagggc gatgccttcc atcagtcgga
     1081 tacacacata ttcatcatca tgggtgcagc gggtgacctg gccaagaaga agatctaccc
     1141 caccatctgg tggctgttcc gggatggcct tctgcccgaa aacaccttca tcgtgggcta
     1201 tgctcagtcc cgcctcacag tggctgacat ccgcaaacag agtgagccct tcttcaaggc
     1261 caccccagag gagaagctca agctggagga cttctttgcc cgcaactcct atgtggctgg
     1321 ccagtacgat gatgcagcct cctaccagcg cctcaacagc cacatgaatg ccctccacct
     1381 ggggtcacag gccaaccgcc tcttctacct ggccttgccc ccgaccgtct acgaggccgt
     1441 caccaagaac attcacgagt cctgcatgag ccagataggc tggaaccgca tcatcgtgga
     1501 gaagcccttc gggagggacc tgcagagctc tgaccggctg tccaaccaca tctcctccct
     1561 gttccgtgag gaccagatct accgcatcga caactacctg ggcacggaga tggtgcagaa
     1621 cctcatggtg ctgagatttg ccaacaggat cttcggcccc atctggaacc gggacaacat
     1681 cgcctgcgtt atcctcacct tcaaggagcc ctttggcact gagggtcgcg ggggctattt
     1741 cgatgaattt gggatcatcc gggacgtgat gcagaaccac ctactgcaga tgctgtgtct
     1801 ggtggccatg gagaagcccg cctccaccaa ctcagatgac gtccgtgatg agaaggtcaa
     1861 ggtgttgaaa tgcatctcag aggtgcaggc caacaatgtg gtcctgggcc agtacgtggg
     1921 gaaccccgat ggagagggcg aggccaccaa agggtacctg gacgacccca cggtgccccg
     1981 cgggtccacc accgccactt ttgcagccgt cgtcctctat gtggagaatg agaggtggga
     2041 tggggtgccc ttcatcctgc gctgcggcaa ggccctgaac gagcgcaagg ccgaggtgag
     2101 gctgcagttc catgatgtgg ccggcgacat cttccaccag cagtgcaagc gcaacgagct
     2161 ggtgatccgc gtgcagccca acgaggccgt gtacaccaag atgatgacca agaagccggg
     2221 catgttcttc aaccccgagg agtcggagct ggacctgacc tacggcaaca gatacaagaa
     2281 cgtgaagctc cctgacgcct atgagcgcct catcctggac gtcttctgcg ggagccagat
     2341 gcacttcgtg cgcagcgacg agctccgtga ggcctggcgt attttcaccc cactgctgca
     2401 ccagattgag ctggagaagc ccaagcccat cccctatatt tatggcagcc gaggccccac
     2461 ggaggcagac gagctgatga agagagtggg tttccagtat gagggcacct acaagtgggt
     2521 gaacccccac aagctcagac cggtcgccac catggtgagc aagggcgagg agctgttcac
     2581 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     2641 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     2701 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgtcct ggggcgtgca
     2761 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     2821 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     2881 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     2941 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactact ttagcgacaa
     3001 cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca
     3061 caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     3121 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtcca agctgagcaa
     3181 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     3241 cactctcggc atggacgagc tgtacaagct cgagtctaga gggcccgttt aaacccgctg
     3301 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3361 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3421 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     3481 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     3541 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     3601 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3661 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3721 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3781 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3841 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3901 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     3961 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     4021 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4081 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4141 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4201 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4261 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     4321 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     4381 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     4441 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4501 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4561 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     4621 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     4681 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4741 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4801 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4861 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     4921 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     4981 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5041 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5101 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5161 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5221 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     5281 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     5341 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     5401 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5461 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     5521 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5581 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5641 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5701 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5761 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5821 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5881 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5941 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6001 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6061 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6121 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6181 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6241 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6301 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6361 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     6421 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6481 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6541 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6601 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6661 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6721 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6781 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6841 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6901 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6961 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7021 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7081 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7141 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7201 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7261 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7321 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7381 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7441 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7501 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7561 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7621 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7681 cgcgcac
//
