LOCUS       sequence-400080-        3732 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Brightness calibration .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3732)
  AUTHORS   Sankaran J, Balasubramanian H, Tang WH, Ng XW, Rollin A, Wohland T
  TITLE     Simultaneous spatiotemporal super-resolution and multi-parametric 
            fluorescence microscopy.
  JOURNAL   Nat Commun. 2021 Mar 19;12(1):1748. doi: 10.1038/s41467-021-22002-9.
  PUBMED    33741958
REFERENCE   2  (bases 1 to 3732)
  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 
            Commun."; date: "2021-03-19"; volume: "12(1):1748. doi"; pages: " 
            10.1038/s41467-021-22002-9"
COMMENT     Sequence Label: PMT-mApple
FEATURES             Location/Qualifiers
     source          1..3732
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..21
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     enhancer        82..385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..589
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     540..560
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    616..640
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      641..650
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             698..1405
                     /codon_start=1
                     /product="photostable monomeric derivative of DsRed (Shaner
                     et al., 2008)"
                     /label=mApple
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEA
                     FQTAKLKVTKGGPLPFAWDILSPQFMYGSKVYIKHPADIPDYFKLSFPEGFRWERVMNF
                     EDGGIIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGWEASEERMYPEDGALK
                     SEIKKRLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     1111..1130
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(1436..1460)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1489..1610
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1500..1519)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1554..1573
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1726..1743)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1897..2485)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1977..1996)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2656..3447,3448..3516))
                     /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     3279..3298
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3517..3621)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3689..3707
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 caactttgta tagaaaagtt gtagttatta atagtaatca attacggggt cattagttca
       61 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      121 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      181 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      361 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      421 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      481 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      541 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
      601 ccgtcagatc accggtcaag tttgtacaaa aaagcaggct gccaccatgg gctgcttctt
      661 cagcaagcgg cggaaggccg acaaggagag cactagtatg gtgagcaagg gcgaggagaa
      721 taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa
      781 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg cctttcagac
      841 cgctaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc
      901 tcagttcatg tacggctcca aggtctacat taagcaccca gccgacatcc ccgactactt
      961 caagctgtcc ttccccgagg gcttcaggtg ggagcgcgtg atgaacttcg aggacggcgg
     1021 cattattcac gttaaccagg actcctccct gcaggacggc gtgttcatct acaaggtgaa
     1081 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg
     1141 ggaggcctcc gaggagcgga tgtaccccga ggacggcgcc ctgaagagcg agatcaagaa
     1201 gaggctgaag ctgaaggacg gcggccacta cgccgccgag gtcaagacca cctacaaggc
     1261 caagaagccc gtgcagctgc ccggcgccta catcgtcgac atcaagttgg acatcgtgtc
     1321 ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac
     1381 cggcggcatg gacgagctgt acaagggtgg cggagggtcc gctagctaaa agcttaccca
     1441 gctttcttgt acaaagtggt gatggccggc cgcttcgagc agacatgata agatacattg
     1501 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     1561 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     1621 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     1681 aaaacctcta caaatgtggt agcggccgcg gcgctcttcc gcttcctcgc tcactgactc
     1741 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     1801 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     1861 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1921 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1981 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2041 taccggatac ctgtccgcct ttctctcttc gggaagcgtg gcgctttctc atagctcacg
     2101 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2161 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2221 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2281 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2341 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2401 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2461 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2521 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2581 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2641 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     2701 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     2761 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     2821 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     2881 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     2941 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3001 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3061 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3121 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3181 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3241 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     3301 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3361 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3421 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     3481 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     3541 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     3601 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     3661 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcggcg
     3721 cgccgcggcc gc
//
