LOCUS       sequence-133683-        6233 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Venus-tagged Apollo-NADP+ in mammalian cells as a reporter
            for cytoplasmic NADP+.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6233)
  AUTHORS   Cameron WD, Bui CV, Hutchinson A, Loppnau P, Graslund S, Rocheleau 
            JV
  TITLE     Apollo-NADP: a spectrally tunable family of genetically encoded 
            sensors for NADP
  JOURNAL   Nat Methods. 2016 Feb 15. doi: 10.1038/nmeth.3764.
  PUBMED    26878383
REFERENCE   2  (bases 1 to 6233)
  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 
            Methods. 2016 Feb 15. doi: 10.1038/nmeth.3764."
COMMENT     Sequence Label: Venus-tagged Apollo-NADP+
FEATURES             Location/Qualifiers
     source          1..6233
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2173..2182
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2179..2181,2182..2184,2185..2898)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2224..2245)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2485..2504)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2832..2853
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3021..3142
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3058..3077)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3112..3131
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3149..3604)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3286..3307)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3498..3517
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3631..3735
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3737..4094
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3737..3928
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3760..3780)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3945..4080
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4129..4923
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4129..4923
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4183..4202)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4793..4812
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5111..5130)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5155..5202
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5531..6119
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6020..6039
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagat ggcagagcag gtggccctga gccggaccca ggtgtgcggg
      661 atcctgcggg aagagctttt ccagggcgat gccttccatc agtcggatac acacatattc
      721 atcatcatgg gtgcagcggg tgacctggcc aagaagaaga tctaccccac catctggtgg
      781 ctgttccggg atggccttct gcccgaaaac accttcatcg tgggctatgc tcagtcccgc
      841 ctcacagtgg ctgacatccg caaacagagt gagcccttct tcaaggccac cccagaggag
      901 aagctcaagc tggaggactt ctttgcccgc aactcctatg tggctggcca gtacgatgat
      961 gcagcctcct accagcgcct caacagccac atgaatgccc tccacctggg gtcacaggcc
     1021 aaccgcctct tctacctggc cttgcccccg accgtctacg aggccgtcac caagaacatt
     1081 cacgagtcct gcatgagcca gataggctgg aaccgcatca tcgtggagaa gcccttcggg
     1141 agggacctgc agagctctga ccggctgtcc aaccacatct cctccctgtt ccgtgaggac
     1201 cagatctacc gcatcgacaa ctacctgggc acggagatgg tgcagaacct catggtgctg
     1261 agatttgcca acaggatctt cggccccatc tggaaccggg acaacatcgc ctgcgttatc
     1321 ctcaccttca aggagccctt tggcactgag ggtcgcgggg gctatttcga tgaatttggg
     1381 atcatccggg acgtgatgca gaaccaccta ctgcagatgc tgtgtctggt ggccatggag
     1441 aagcccgcct ccaccaactc agatgacgtc cgtgatgaga aggtcaaggt gttgaaatgc
     1501 atctcagagg tgcaggccaa caatgtggtc ctgggccagt acgtggggaa ccccgatgga
     1561 gagggcgagg ccaccaaagg gtacctggac gaccccacgg tgccccgcgg gtccaccacc
     1621 gccacttttg cagccgtcgt cctctatgtg gagaatgaga ggtgggatgg ggtgcccttc
     1681 atcctgcgct gcggcaaggc cctgaacgag cgcaaggccg aggtgaggct gcagttccat
     1741 gatgtggccg gcgacatctt ccaccagcag tgcaagcgca acgagctggt gatccgcgtg
     1801 cagcccaacg aggccgtgta caccaagatg atgaccaaga agccgggcat gttcttcaac
     1861 cccgaggagt cggagctgga cctgacctac ggcaacagat acaagaacgt gaagctccct
     1921 gacgcctatg agcgcctcat cctggacgtc ttctgcggga gccagatgca cttcgtgcgc
     1981 agcgacgagc tccgtgaggc ctggcgtatt ttcaccccac tgctgcacca gattgagctg
     2041 gagaagccca agcccatccc ctatatttat ggcagccgag gccccacgga ggcagacgag
     2101 ctgatgaaga gagtgggttt ccagtatgag ggcacctaca agtgggtgaa cccccacaag
     2161 ctcagaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     2221 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     2281 gagggcgatg ccacctacgg caagctgacc ctgaagctga tctgcaccac cggcaagctg
     2341 cccgtgccct ggcccaccct cgtgaccacc ctgggctacg gcctgcagtg cttcgcccgc
     2401 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     2461 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     2521 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     2581 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcacc
     2641 gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac
     2701 ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     2761 ctgctgcccg acaaccacta cctgagctac cagtccaaac tgagcaaaga ccccaacgag
     2821 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     2881 gacgagctgt acaagtaaag cggccgcgac tctagatcat aatcagccat accacatttg
     2941 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     3001 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     3061 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     3121 ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc
     3181 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     3241 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     3301 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     3361 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     3421 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     3481 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     3541 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     3601 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     3661 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     3721 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     3781 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3841 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3901 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3961 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4021 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4081 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4141 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4201 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4261 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     4321 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4381 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4441 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4501 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4561 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4621 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     4681 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4741 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4801 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4861 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4921 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     4981 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5041 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     5101 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     5161 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     5221 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     5281 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     5341 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     5401 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5461 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5521 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5581 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5641 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     5701 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5761 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5821 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5881 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5941 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6001 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6061 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6121 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6181 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
