LOCUS       sequence-380426-        7708 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mTurq2-Apollo-NADP+ sensor localized to the nucleus.
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: Nuc-mTurq2-Apollo-NADP+
FEATURES             Location/Qualifiers
     source          1..7708
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(54..73)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        245..624
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        625..828
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     779..799
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     873..892
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        873..891
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             974..994
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     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..3271)
                     /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(3309..3326)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3315..3539
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3585..4013
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3672..3691)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3882..3903
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4022..4042)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4027..4356
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4207..4342
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4269..4288
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4423..5217
                     /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(4477..4496)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5087..5106
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5391..5512
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5428..5447)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5482..5501
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5561..5577)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5561..5577)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5574..5596)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5585..5601
                     /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(5609..5615,5616..5633,5634..5639))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5654..5675
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5792..5809)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5963..6551)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6043..6062)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6722..7513,7514..7582))
                     /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     7345..7364
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7583..7687)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc
       61 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
      121 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
      181 aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg
      241 cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
      301 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      361 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      421 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      481 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      541 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      601 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      661 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      721 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      781 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact
      841 agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa
      901 gctggctagc gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt
      961 ggaattcatg gctccaaaga agaagcgtaa ggtagcagag 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 tgtacaagta actcgagtct agagggcccg tttaaacccg
     3301 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3361 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3421 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3481 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     3541 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3601 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3661 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3721 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3781 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3841 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3901 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3961 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     4021 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4081 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4141 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4201 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4261 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4321 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4381 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4441 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4501 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4561 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4621 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4681 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4741 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4801 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4861 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4921 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4981 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5041 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5101 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5161 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5221 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5281 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5341 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5401 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5461 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5521 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5581 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5641 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5701 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5761 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5821 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5881 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5941 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6001 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6061 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6121 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6181 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6241 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6301 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6361 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6421 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6481 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6541 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6601 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6661 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6721 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6781 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6841 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6901 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6961 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7021 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7081 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7141 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7201 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7261 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7321 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7381 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7441 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7501 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7561 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7621 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7681 ttccgcgcac atttccccga aaagtgcc
//
