LOCUS       sequence-176625-        4060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cyan color variant of bright luminescent protein for bacterial 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4060)
  AUTHORS   Suzuki K, Kimura T, Shinoda H, Bai G, Daniels MJ, Arai Y, Nakano M, 
            Nagai T
  TITLE     Five colour variants of bright luminescent protein for real-time 
            multicolour bioimaging.
  JOURNAL   Nat Commun. 2016 Dec 14;7:13718. doi: 10.1038/ncomms13718.
  PUBMED    27966527
REFERENCE   2  (bases 1 to 4060)
  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 
            Commun."; date: "2016-12-14"; volume: "7:13718. doi"; pages: " 
            10.1038/ncomms13718"
COMMENT     Sequence Label: CeNL_pRSETb
FEATURES             Location/Qualifiers
     source          1..4060
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     52..69
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     154..173
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        154..172
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             204..226
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             246..263
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             267..299
                     /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(303..311,312..326)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(333..335,336..338,339..851)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(378..399)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(639..658)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     986..1007
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1595..1613)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1609..1656
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1753..2208
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1840..1859)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2050..2071
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2234..2338
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2339..2407,2408..3199)
                     /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(2557..2576)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3370..3958
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3859..3878
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
       61 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
      121 attcattaat gcaggatctc gatcccgcga aattaatacg actcactata gggagaccac
      181 aacggtttcc ctctagaaat aattttgttt aactttaaga aggagatata catatgcggg
      241 gttctcatca tcatcatcat catggtatgg ctagcatgac tggtggacag caaatgggtc
      301 gggatctgta cgacgatgac gataaggatc cgatggtgag caagggcgag gagctgttca
      361 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
      421 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
      481 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgtcc tggggcgtgc
      541 agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
      601 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
      661 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
      721 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac tttagcgaca
      781 acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc
      841 acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
      901 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc aagctgagca
      961 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccgggt
     1021 tgcatacact cgaagatttc gttggggact ggcgacagac agccggctac aacctggacc
     1081 aagtccttga acagggaggt gtgtccagtt tgtttcagaa tctcggggtg tccgtaactc
     1141 cgatccaaag gattgtcctg agcggtgaaa atgggctgaa gatcgacatc catgtcatca
     1201 tcccgtatga aggtctgagc ggcgaccaaa tgggccagat cgaaaaaatt tttaaggtgg
     1261 tgtaccctgt ggatgatcat cactttaagg tgatcctgca ctatggcaca ctggtaatcg
     1321 acggggttac gccgaacatg atcgactatt tcggacggcc gtatgaaggc atcgccgtgt
     1381 tcgacggcaa aaagatcact gtaacaggga ccctgtggaa cggcaacaaa attatcgacg
     1441 agcgcctgat caaccccgac ggctccctgc tgttccgagt aaccatcaac ggagtgaccg
     1501 gctggcggct gtgcgaacgc attctggcgt aagaattcga agcttgatcc ggctgctaac
     1561 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     1621 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
     1681 tctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     1741 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     1801 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     1861 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     1921 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     1981 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2041 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     2101 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     2161 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
     2221 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     2281 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     2341 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     2401 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     2461 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     2521 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     2581 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     2641 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     2701 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     2761 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     2821 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     2881 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     2941 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     3001 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     3061 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     3121 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     3181 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     3241 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     3301 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3361 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3421 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3481 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     3541 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3601 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3661 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3721 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     3781 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     3841 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3901 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     3961 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     4021 ctttcctgcg ttatcccctg attctgtgga taaccgtatt
//
