LOCUS       sequence-161607-        3618 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of CyRFP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3618)
  AUTHORS   Laviv T, Kim BB, Chu J, Lam AJ, Lin MZ, Yasuda R
  TITLE     Simultaneous dual-color fluorescence lifetime imaging with novel 
            red-shifted fluorescent proteins.
  JOURNAL   Nat Methods. 2016 Oct 31. doi: 10.1038/nmeth.4046.
  PUBMED    27798609
REFERENCE   2  (bases 1 to 3618)
  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 
            Methods. 2016 Oct 31. doi: 10.1038/nmeth.4046."
COMMENT     Sequence Label: pNCS-CyRFP1
FEATURES             Location/Qualifiers
     source          1..3618
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             164..181
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             185..217
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(221..229,230..244)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             251..952
                     /codon_start=1
                     /product="cyan-excitable red fluorescent protein (Laviv et 
                     al., 2016)"
                     /label=CyRFP1
                     /translation="MVSKGEELIKENMRSKLYLEGSVNGHQFKCTHEGEGKPYEGKQTN
                     RIKVVEGGPLPFAFDILATMFMYGSKVFIKYPADLPDYFKQSFPEGFTWERVMVFEDGG
                     VLTATQDTSLQDGELIYNVKLRGVNFPANGPVMQKKTLGWEPSTETMYPADGGLEGRCD
                     KALKLVGGGHLHVNFKTTYKSKKPVKMPGVHYVDRRLERIKEADNETYVEQYEHAVARY
                     SNLGGGMDELYK"
     primer_bind     complement(1019..1037)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1033..1080
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1177..1632
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1264..1283)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1474..1495
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1658..1762
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1763..1831,1832..2623)
                     /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     complement(1981..2000)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2794..3382
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3283..3302
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3536..3553
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaatcaccg agtttattct tgacacctga tgcgatgaat gatataatag gaaagtactg
      121 ttttgtttaa ctttaagaag gggatataca tatgcggggt tctcatcatc atcatcatca
      181 tggtatggct agcatgactg gtggacagca aatgggtcgg gatctgtacg acgatgacga
      241 taaggatccg atggtgagca agggcgagga gctgatcaag gagaacatga gaagcaagct
      301 gtacctggaa ggcagcgtga acggccacca gttcaagtgc acccacgaag gggagggcaa
      361 gccctacgag ggcaagcaga ccaacaggat caaggtggtg gagggaggcc ccctgccgtt
      421 cgcattcgac atcctggcca ccatgtttat gtacgggagc aaggtgttca tcaagtaccc
      481 cgccgacctc cccgattatt ttaagcagtc cttccctgag ggcttcacat gggagagagt
      541 catggtgttc gaagacgggg gcgtgctgac cgccacccag gacaccagcc tccaggacgg
      601 cgagctcatc tacaacgtca agctcagagg ggtgaacttc ccagccaacg gccccgtgat
      661 gcagaagaaa acactgggct gggagcccag caccgagacc atgtaccccg ctgacggcgg
      721 cctggaaggc agatgcgaca aggccctgaa gctcgtgggc gggggccacc tgcacgtcaa
      781 cttcaagacc acatacaagt ccaagaaacc cgtgaagatg cccggcgtcc actacgtgga
      841 ccgcagactg gaaagaatca aggaggccga caacgagacc tacgtcgagc agtacgagca
      901 cgctgtggcc agatactcca acctgggcgg aggcatggac gagctgtaca agtaatgaat
      961 tcgaagcttg atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc
     1021 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     1081 ctgaaaggag gaactatatc cggatctggc gtaatagcga agaggcccgc accgatcgcc
     1141 cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa
     1201 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     1261 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     1321 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     1381 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     1441 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     1501 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     1561 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     1621 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     1681 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     1741 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     1801 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     1861 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     1921 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     1981 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     2041 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     2101 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     2161 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     2221 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     2281 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     2341 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     2401 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     2461 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     2521 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     2581 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     2641 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     2701 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     2761 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2821 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2881 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2941 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3001 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3061 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3121 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3181 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3241 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3301 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3361 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3421 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3481 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     3541 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     3601 gccgattcat taatgcag
//
