LOCUS       sequence-490399-        6263 bp    DNA     circular SYN 23-OCT-2025
DEFINITION  Fluorescent reporter for lactate (with lifetime and intensity 
            response); expresses in mammalian cells and can be packaged as AAV.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6263)
  AUTHORS   Koveal D, Rosen PC, Meyer DJ, Diaz-Garcia CM, Wang Y, Cai LH, Chou 
            PJ, Weitz DA, Yellen G
  TITLE     A high-throughput multiparameter screen for accelerated development 
            and optimization of soluble genetically encoded fluorescent 
            biosensors.
  JOURNAL   Nat Commun. 2022 May 25;13(1):2919. doi: 10.1038/s41467-022-30685-x.
  PUBMED    35614105
REFERENCE   2  (bases 1 to 6263)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 23, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2022-05-25"; volume: "13(1):2919. doi"; pages: " 
            10.1038/s41467-022-30685-x"
COMMENT     Sequence Label: pAAV-CAG-LiLac
FEATURES             Location/Qualifiers
     source          1..6263
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   19..148
                     /label=AAV2 ITR
     promoter        459..736
                     /label=chicken beta-actin promoter
     primer_bind     1015..1037
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1099..1118
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1160..1180
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     primer_bind     complement(1226..1247)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1487..1506)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2456..2704
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
                     YLSTQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELY"
     misc_feature    2725..3313
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2778..2798)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3196..3207)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3356..3477
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3367..3386)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3421..3440
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3639..3768
                     /label=AAV2 ITR
     rep_origin      complement(4005..4593)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4085..4104)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4764..5555,5556..5624))
                     /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     5387..5406
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5625..5729)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5755..6210)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5892..5913)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6104..6123
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tcttcgctat tacgccagct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
       61 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
      121 gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta
      181 tctacgtagc catgctctag gaagagtacc attgacgtca ataatgacgt atgttcccat
      241 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      301 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      361 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      421 gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg
      481 cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta
      541 attattttgt gcagcgatgg gggcgggggg gggggggggg cgcgcgccag gcggggcggg
      601 gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg
      661 cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg
      721 aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc
      781 cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg
      841 ggacggccct tctcctccgg gctgtaatta gcgcttggtt taatgacggc ttgtttcttt
      901 tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg gggggagcgg
      961 ctcggggctg tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg
     1021 gcttctggcg tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct
     1081 ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag
     1141 aattggatcc accatgaaac atcaccatca ccatcaccat ggagtgagca agggcgagga
     1201 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     1261 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     1321 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgtcctg
     1381 gggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     1441 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     1501 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     1561 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactcctc
     1621 cggccgcacc ggcattgacc cctttactga aagcgtgttg caaagccaag ccacagaatt
     1681 gctgcaaaaa aaagctcagt tagtcagttt taaaattcaa ggcattatga agcgcatttt
     1741 tatgggcgct aacacccttg aaaaattttt aagcgatgaa aatagcgcta tcaacgacac
     1801 cctcaaacgc cgcatgctct ctgagttttt gttagcaaac cctcatgtgt tattggttag
     1861 cgcgatttat acgaataata atgaacgtgt catcactgcc atgagcatgg attcaaaaat
     1921 cgcctaccct aataccacgc tcaatgaaaa catgaccaat caaatccgtt cgctcaaaag
     1981 tattacccat tcagatccct attataaaga ggttaatggc gataaaatct atggcatgga
     2041 tattaccctc ccccttatgg gtaagaatca aaatgctatt ggcgcgctga atttcttttt
     2101 aaacattgac gctttttata ccgatgtggt aggcaagaaa aagagcaaca cctttttaat
     2161 ggggaaagac ggccgtcttt taatcaaccc taatcgtgag atccaagata agattttaag
     2221 cgctatcaat ccagatcgcc gtgtcgctaa agctgtggag tattacaatc aaaacgaagc
     2281 gggcactttg agctaccatt cattgagcgg gaatacagaa acctttttag ccattcagcc
     2341 ctttgatttt tttgaagaaa aagggaataa cggcaatcat tggcgttggg caattgggaa
     2401 atatgtcaat aaatctttag tcttttccag ccacagcaac gtctatatca ccgccgacaa
     2461 gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcggcgt
     2521 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     2581 cgacaaccac tacctgagca cccagtccaa gctgagcaaa gaccccaacg agaagcgcga
     2641 tcacatggtc ctactggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     2701 gtaccagtga tttccggaat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     2761 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     2821 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     2881 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     2941 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     3001 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     3061 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     3121 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     3181 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     3241 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     3301 gccgcctccc cgcatcgata ccgtcgaccc gggcggccgc ttcgagcaga catgataaga
     3361 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     3421 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     3481 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa
     3541 agcaagtaaa acctctacaa atgtggtaaa atcgataagg atcttcctag agcatggcta
     3601 cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt
     3661 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     3721 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcattaatga
     3781 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     3841 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3901 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3961 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4021 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4081 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4141 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4201 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4261 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4321 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4381 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4441 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4501 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4561 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4621 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4681 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4741 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4801 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4861 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4921 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4981 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5041 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     5101 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     5161 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     5221 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     5281 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     5341 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     5401 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     5461 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     5521 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     5581 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     5641 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5701 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg
     5761 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     5821 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     5881 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     5941 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     6001 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     6061 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     6121 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     6181 ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt
     6241 gggaagggcg atcggtgcgg gcc
//
