LOCUS       sequence-223054-        5393 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of tgRFP labelled LOV2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5393)
  AUTHORS   Reis JM, Xu X, McDonald S, Woloschuk RM, Jaikaran ASI, Vizeacoumar 
            FS, Woolley GA, Uppalapati M
  TITLE     Discovering Selective Binders for Photoswitchable Proteins Using 
            Phage Display.
  JOURNAL   ACS Synth Biol. 2018 Oct 19;7(10):2355-2364. doi: 
            10.1021/acssynbio.8b00123. Epub 2018 Sep 27.
  PUBMED    30203962
REFERENCE   2  (bases 1 to 5393)
  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: 
            "10.1021/acssynbio.8b00123"; journalName: "ACS Synth Biol"; date: 
            "2018-10-19- 19"; volume: "7"; issue: "10"; pages: "2355-2364"
COMMENT     Sequence Label: PLL7.0 tgRFP LOV
FEATURES             Location/Qualifiers
     source          1..5393
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        618..821
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      861..870
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             867..1598
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     protein_bind    complement(2195..2228)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2284..2870
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2337..2357)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2755..2766)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2873..2889)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2874..2890)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             3399..3579
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(3641..4229)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3721..3740)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4400..5191,5192..5260))
                     /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     5023..5042
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5261..5365)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gagcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataattacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
      841 tccgctagaa ttgaccggtc gccaccatgg tgtctaaggg cgaagagctg attaaggaga
      901 acatgcacat gaagctgtac atggagggca ccgtgaacaa ccaccacttc aagtgcacat
      961 ccgagggcga aggcaagccc tacgagggca cccagaccat gagaatcaag gtggtcgagg
     1021 gcggccctct ccccttcgcc ttcgacatcc tggctaccag cttcatgtac ggcagcagaa
     1081 ccttcatcaa ccacacccag ggcatccccg acttctttaa gcagtccttc cctgagggct
     1141 tcacatggga gagagtcacc acatacgaag acgggggcgt gctgaccgct acccaggaca
     1201 ccagcctcca ggacggctgc ctcatctaca acgtcaagat cagaggggtg aacttcccat
     1261 ccaacggccc tgtgatgcag aagaaaacac tcggctggga ggccaacacc gagatgctgt
     1321 accccgctga cggcggcctg gaaggcagaa ccgacatggc cctgaagctc gtgggcgggg
     1381 gccacctgat ctgcaacttc aagaccacat acagatccaa gaaacccgct aagaacctca
     1441 agatgcccgg cgtctactat gtggaccaca gactggaaag aatcaaggag gccgacaaag
     1501 agacctacgt cgagcagcac gaggtggctg tggccagata ctgcgacctc cctagcaaac
     1561 tggggcacaa acttaatggc atggacgagc tgtacaagtc cggactcaga tctcgagctc
     1621 aagcttcgaa cgaattcggc attgatctga gcggcctgac cctgcaggcc gccctggcca
     1681 caacactgga gaggatcgag aagaacttca ttattacaga tcctagactg cccgataacc
     1741 ctattatttt tgccagcgat agttttctgc agctgacaga gtacagcagg gaggagattt
     1801 tgggaaggaa ctgcagattt ctgcaggggc cagagacaga tagagccaca gtgaggaaga
     1861 ttagagatgc cattgataac cagactgagg tgacagtgca gctgattaac tacacaaaga
     1921 gcgggaaaaa gttttggaac ctgtttcatc tgcagcctat gagagatcag aagggagatg
     1981 tgcagtactt tattggcgtg cagttggatg gaacagagca tgtgagagat gccgctgaga
     2041 gggaggccgt gatgctgatt aagaagacag ccgaggagat tgatgaggcc gctaaggagc
     2101 tgcccgatgc taacctgaga cctgaggatc tgtgggccaa ccactaaggt ccaccggatc
     2161 aagataacct gcaggcaatt cgtcgaggga cctaataact tcgtatagca tacattatac
     2221 gaagttatac atgtttaagg gttccggttc cactaggtac aattcgatat caagcttatc
     2281 gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     2341 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     2401 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     2461 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     2521 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     2581 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     2641 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     2701 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     2761 ctcaatccag cggaccttcc ttcccggcct gctgccggct ctgcggcctc ttccgcgtct
     2821 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     2881 tcgacctcga tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta
     2941 ccaatgctga ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca
     3001 cacctcaggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     3061 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg
     3121 atctgtggat ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc
     3181 cagggatcag atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag
     3241 agaaggtaga agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc
     3301 atgggatgga tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat
     3361 ttcatcacat ggcccgagag ctgcatccgg actgtactgg gtctctctgg ttagaccaga
     3421 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     3481 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     3541 ccctcagacc cttttagtca gtgtggaaaa tctctagcag catgtgagca aaaggccagc
     3601 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3661 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3721 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3781 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3841 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3901 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3961 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4021 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4081 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4141 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4201 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4261 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4321 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4381 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4441 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4501 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4561 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4621 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4681 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4741 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4801 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4861 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4921 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4981 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5041 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5101 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5161 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5221 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     5281 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5341 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gac
//
