LOCUS       sequence-223056-        5322 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of tgRFP labelled cPYP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5322)
  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 5322)
  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 cPYP
FEATURES             Location/Qualifiers
     source          1..5322
                     /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(2124..2157)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2213..2799
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2266..2286)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2684..2695)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2802..2818)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2803..2819)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             3328..3508
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(3570..4158)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3650..3669)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4329..5120,5121..5189))
                     /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     4952..4971
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5190..5294)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa 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 ataatgacgt 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 cctgcagatg aagggggata
     1681 gctactgggt gtttgtgaag agagtgggag gaagcggagg gagtggagga atggagcatg
     1741 tggcctttgg aagtgaggat attgagaaca cattggccaa gatggatgat gggcagctgg
     1801 atggactggc ctttggagcc attcagctgg atggagatgg gaacatcctg cagtacaacg
     1861 ccgccgaggg ggacatcaca ggccgggacc ccaagcaggt catcggcaag aacttcttca
     1921 aggacgtggc cccctgcaca gatagccccg agttctacgg caagtttaag gagggggtgg
     1981 ccagcggcaa cctgaacaca atgttcgagt acaccttcga ctaccaggag acacccacca
     2041 aggtgaaggt gcacatgaag aaggccctga gctaaggatc caccggatca agataacctg
     2101 caggcaattc gtcgagggac ctaataactt cgtatagcat acattatacg aagttataca
     2161 tgtttaaggg ttccggttcc actaggtaca attcgatatc aagcttatcg ataatcaacc
     2221 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     2281 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     2341 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     2401 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     2461 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     2521 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     2581 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     2641 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     2701 ggaccttcct tcccggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     2761 ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacctcgat
     2821 cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat
     2881 tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta
     2941 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     3001 gggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc
     3061 taccacacac aaggctactt ccctgattgg cagaactaca caccagggcc agggatcaga
     3121 tatccactga cctttggatg gtgctacaag ctagtaccag ttgagcaaga gaaggtagaa
     3181 gaagccaatg aaggagagaa cacccgcttg ttacaccctg tgagcctgca tgggatggat
     3241 gacccggaga gagaagtatt agagtggagg tttgacagcc gcctagcatt tcatcacatg
     3301 gcccgagagc tgcatccgga ctgtactggg tctctctggt tagaccagat ctgagcctgg
     3361 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     3421 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     3481 ttttagtcag tgtggaaaat ctctagcagc atgtgagcaa aaggccagca aaaggccagg
     3541 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3601 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3661 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3721 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3781 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3841 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3901 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3961 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4021 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4081 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4141 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4201 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4261 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4321 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4381 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4441 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4501 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4561 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4621 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4681 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4741 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4801 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4861 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4921 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4981 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5041 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5101 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5161 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5221 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5281 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg ac
//
