LOCUS       sequence-180855-        3706 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of a fluorescent protein, Yellow Fluorescent 
            Protein (YFP).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3706)
  AUTHORS   Cubitt AB, Heim R, Adams SR, Boyd AE, Gross LA, Tsien RY
  TITLE     Understanding, improving and using green fluorescent proteins.
  JOURNAL   Trends Biochem Sci. 1995 Nov;20(11):448-55.
  PUBMED    8578587
REFERENCE   2  (bases 1 to 3706)
  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: "Trends 
            Biochem Sci."; date: "1995-11"; volume: "20(11)"; pages: "448-55"
COMMENT     Sequence Label: pNCS YFP
FEATURES             Location/Qualifiers
     source          1..3706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     57..74
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     159..178
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        159..177
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             261..283
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             303..320
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             324..356
                     /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(360..368,369..383)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(390..392,393..395,396..1109)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(435..456)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(696..715)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1043..1064
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1246..1264)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1260..1307
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1404..1859
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1491..1510)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1701..1722
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1885..1989
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1990..2058,2059..2850)
                     /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(2208..2227)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3021..3609
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3510..3529
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
       61 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
      121 ggccgattca ttaatgcagg atctcgatcc cgcgaaatta atacgactca ctatagggag
      181 accacaacgg tttccctcta gaatcaccga gtttattctt gacacctgat gcgatgaatg
      241 atataatagg aaagtactgt tttgtttaac tttaagaagg agatatacat atgcggggtt
      301 ctcatcatca tcatcatcat ggtatggcta gcatgactgg tggacagcaa atgggtcggg
      361 atctgtacga cgatgacgat aaggatccga tggtgagcaa gggcgaggag ctgttcaccg
      421 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
      481 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagctg atctgcacca
      541 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgggctac ggcctgcagt
      601 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
      661 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
      721 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
      781 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
      841 tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca
      901 acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
      961 acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc ctgagcaaag
     1021 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     1081 ctctcggcat ggacgagctg tacaagtaag aattcttact tgtacagctc gcccttgctc
     1141 accatcggat ccgatggtga gcatggacga gctgtacaag taagaattcg aagcttgatc
     1201 cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac
     1261 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa
     1321 ctatatccgg atctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     1381 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     1441 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     1501 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     1561 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     1621 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     1681 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     1741 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     1801 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     1861 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     1921 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     1981 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     2041 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     2101 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     2161 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     2221 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     2281 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     2341 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     2401 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     2461 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     2521 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     2581 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     2641 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     2701 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     2761 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     2821 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     2881 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     2941 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     3001 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3061 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3121 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     3181 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3241 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3301 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3361 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3421 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     3481 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3541 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     3601 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     3661 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataacc
//
