LOCUS       sequence-400078-        3720 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 3720)
  AUTHORS   Balasubramanian H, Sankaran J, Pandey S, Goh CJH, Wohland T
  TITLE     The dependence of EGFR oligomerization on environment and structure:
            A camera-based N
  JOURNAL   Biophys J. 2022 Dec 6;121(23):4452-4466. doi: 
            10.1016/j.bpj.2022.11.003. Epub 2022 Nov 5.
  PUBMED    36335429
REFERENCE   2  (bases 1 to 3720)
  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"; doi: 
            "10.1016/j.bpj.2022.11.003"; journalName: "Biophys J"; date: 
            "2022-12-6- 6"; volume: "121"; issue: "23"; pages: "4452-4466"
COMMENT     Sequence Label: PMT-mScarlet
FEATURES             Location/Qualifiers
     source          1..3720
                     /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..1393
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(831..849)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1099..1118
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(1424..1448)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1477..1598
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1488..1507)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1542..1561
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1714..1731)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1885..2473)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1965..1984)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2644..3435,3436..3504))
                     /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     3267..3286
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3505..3609)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3677..3695
                     /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 gcgaggcagt
      721 gatcaaggag ttcatgcggt tcaaggtgca catggagggc tccatgaacg gccacgagtt
      781 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
      841 ggtgaccaag ggtggccccc tgcccttctc ctgggacatc ctgtcccctc agttcatgta
      901 cggctccagg gccttcacca agcaccccgc cgacatcccc gactactata agcagtcctt
      961 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg ccgtgaccgt
     1021 gacccaggac acctccctgg aggacggcac cctgatctac aaggtgaagc tccgcggcac
     1081 caacttccct cctgacggcc ccgtaatgca gaagaagaca atgggctggg aagcgtccac
     1141 cgagcggttg taccccgagg acggcgtgct gaagggcgac attaagatgg ccctgcgcct
     1201 gaaggacggc ggccgctacc tggcggactt caagaccacc tacaaggcca agaagcccgt
     1261 gcagatgccc ggcgcctaca acgtcgaccg caagttggac atcacctccc acaacgagga
     1321 ctacaccgtg gtggaacagt acgaacgctc cgagggccgc cactccaccg gcggcatgga
     1381 cgagctgtac aagggtggcg gagggtccgc tagctaaaag cttacccagc tttcttgtac
     1441 aaagtggtga tggccggccg cttcgagcag acatgataag atacattgat gagtttggac
     1501 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     1561 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     1621 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     1681 aatgtggtag cggccgcggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     1741 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     1801 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     1861 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1921 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     1981 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2041 gtccgccttt ctctcttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2101 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2161 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2221 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2281 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     2341 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     2401 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     2461 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     2521 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     2581 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     2641 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     2701 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     2761 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     2821 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     2881 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     2941 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     3001 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     3061 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     3121 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     3181 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     3241 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     3301 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     3361 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     3421 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     3481 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     3541 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     3601 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     3661 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcggcgcg ccgcggccgc
//
