LOCUS       sequence-401132-        4437 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 4437)
  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 4437)
  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_2
FEATURES             Location/Qualifiers
     source          1..4437
                     /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"
     CDS             1415..2110
                     /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"
     protein_bind    complement(2141..2165)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2194..2315
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2205..2224)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2259..2278
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2431..2448)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2602..3190)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2682..2701)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3361..4152,4153..4221))
                     /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     3984..4003
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4222..4326)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4394..4412
                     /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 tagtatggtg agcaagggcg aggcagtgat
     1441 caaggagttc atgcggttca aggtgcacat ggagggctcc atgaacggcc acgagttcga
     1501 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     1561 gaccaagggt ggccccctgc ccttctcctg ggacatcctg tcccctcagt tcatgtacgg
     1621 ctccagggcc ttcaccaagc accccgccga catccccgac tactataagc agtccttccc
     1681 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgccg tgaccgtgac
     1741 ccaggacacc tccctggagg acggcaccct gatctacaag gtgaagctcc gcggcaccaa
     1801 cttccctcct gacggccccg taatgcagaa gaagacaatg ggctgggaag cgtccaccga
     1861 gcggttgtac cccgaggacg gcgtgctgaa gggcgacatt aagatggccc tgcgcctgaa
     1921 ggacggcggc cgctacctgg cggacttcaa gaccacctac aaggccaaga agcccgtgca
     1981 gatgcccggc gcctacaacg tcgaccgcaa gttggacatc acctcccaca acgaggacta
     2041 caccgtggtg gaacagtacg aacgctccga gggccgccac tccaccggcg gcatggacga
     2101 gctgtacaag ggtggcggag ggtccgctag ctaaaagctt acccagcttt cttgtacaaa
     2161 gtggtgatgg ccggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa
     2221 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     2281 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     2341 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     2401 gtggtagcgg ccgcggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2461 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2521 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2581 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2641 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2701 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2761 cgcctttctc tcttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     2821 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2881 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     2941 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3001 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3061 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3121 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3181 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3241 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3301 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3361 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3421 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3481 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3541 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3601 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3661 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3721 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     3781 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     3841 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     3901 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     3961 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4021 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4081 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4141 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4201 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4261 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4321 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     4381 attaacctat aaaaataggc gtatcacgag gccctttcgt cggcgcgccg cggccgc
//
