LOCUS       sequence-208243-        3574 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes for the fluorescent protein CFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3574)
  AUTHORS   Sarabipour S, King C, Hristova K
  TITLE     Uninduced high-yield bacterial expression of fluorescent proteins.
  JOURNAL   Anal Biochem. 2014 Mar 15;449:155-7. doi: 10.1016/j.ab.2013.12.027. 
            Epub 2013 Dec 28.
  PUBMED    24378720
REFERENCE   2  (bases 1 to 3574)
  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: "Anal 
            Biochem."; date: "2014-03-15"; volume: "449:155-7. doi"; pages: " 
            10.1016/j.ab.2013.12.027. Epub 2013 Dec 28"
COMMENT     Sequence Label: pRSET-B CFP
FEATURES             Location/Qualifiers
     source          1..3574
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      17..605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     506..525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     759..776
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     861..880
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        861..879
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             911..933
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             953..970
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             974..1006
                     /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(1010..1018,1019..1033)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1040..1753
                     /codon_start=1
                     /product="cyan variant of GFP"
                     /label=CFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKAH
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     primer_bind     complement(1079..1100)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1340..1359)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1687..1708
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1816..1834)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1830..1877
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1974..2429
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2061..2080)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2271..2292
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2455..2559
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2560..2628,2629..3420)
                     /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(2778..2797)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
       61 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      121 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      181 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      241 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      301 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      361 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      421 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      481 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      541 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      601 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      661 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
      721 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
      781 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
      841 ggatctcgat cccgcgaaat taatacgact cactataggg agaccacaac ggtttccctc
      901 tagaaataat tttgtttaac tttaagaagg agatatacat atgcggggtt ctcatcatca
      961 tcatcatcat ggtatggcta gcatgactgg tggacagcaa atgggtcggg atctgtacga
     1021 cgatgacgat aaggatccaa gcaagggcga ggagctgttc accggggtgg tgcccatcct
     1081 ggtcgagctg gacggcgacg taaacggcca caggttcagc gtgtccggcg agggcgaggg
     1141 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     1201 gccctggccc accctcgtga ccaccctgac ctggggcgtg cagtgcttca gccgctaccc
     1261 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     1321 gcgtaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     1381 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     1441 catcctgggg cacaagctgg agtacaacta catcagccac aacgtctata tcaccgccga
     1501 caagcagaag aacggcatca aggcccactt caagatccgc cacaacatcg aggacggcag
     1561 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     1621 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     1681 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     1741 gctgtacaag taagaattcg aagcttgatc cggctgctaa caaagcccga aaggaagctg
     1801 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
     1861 tcttgagggg ttttttgctg aaaggaggaa ctatatccgg atctggcgta atagcgaaga
     1921 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
     1981 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2041 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2101 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2161 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     2221 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     2281 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     2341 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     2401 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     2461 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2521 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     2581 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     2641 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     2701 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     2761 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     2821 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2881 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2941 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     3001 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     3061 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     3121 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     3181 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     3241 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3301 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     3361 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     3421 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     3481 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3541 ttttcgttcc actgagcgtc agaccccgta gaaa
//
