LOCUS       sequence-194938-        3593 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the fluorescent protein meleCFP for constitutive expression 
            in E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3593)
  AUTHORS   Molina RS, Tran TM, Campbell RE, Lambert GG, Salih A, Shaner NC, 
            Hughes TE, Drobizhev M
  TITLE     Blue-Shifted Green Fluorescent Protein Homologues Are Brighter than 
            Enhanced Green Fluorescent Protein under Two-Photon Excitation.
  JOURNAL   J Phys Chem Lett. 2017 Jun 15;8(12):2548-2554. doi: 
            10.1021/acs.jpclett.7b00960. Epub 2017 May 25.
  PUBMED    28530831
REFERENCE   2  (bases 1 to 3593)
  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/acs.jpclett.7b00960"; journalName: "J Phys Chem Lett"; 
            date: "2017-06-15- 15"; volume: "8"; issue: "12"; pages: "2548-2554"
COMMENT     Sequence Label: pRSM13
FEATURES             Location/Qualifiers
     source          1..3593
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(61..79)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      75..122
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      219..674
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(306..325)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     516..537
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        700..804
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(805..873,874..1665)
                     /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(1023..1042)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1836..2424
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2325..2344
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2578..2595
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2680..2699
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2680..2698
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2782..2804
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2824..2841
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2845..2877
                     /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(2881..2889,2890..2904)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
ORIGIN
        1 attcgaagct tgatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
       61 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
      121 tgctgaaagg aggaactata tccggatctg gcgtaatagc gaagaggcca gcaccgatcg
      181 cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt
      241 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
      301 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
      361 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
      421 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
      481 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
      541 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
      601 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
      661 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
      721 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
      781 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
      841 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
      901 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
      961 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     1021 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     1081 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     1141 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     1201 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     1261 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     1321 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     1381 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     1441 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     1501 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     1561 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     1621 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     1681 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     1741 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     1801 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     1861 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     1921 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     1981 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     2041 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     2101 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     2161 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     2221 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     2281 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     2341 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     2401 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     2461 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     2521 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     2581 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     2641 tggccgattc attaatgcag gatctcgatc ccgcgaaatt aatacgactc actataggga
     2701 gaccacaacg gtttccctct agaatcaccg agtttattct tgacacctga tgcgatgaat
     2761 gatataatag gaaagtactg ttttgtttaa ctttaagaag gagatataca tatgcggggt
     2821 tctcatcatc atcatcatca tggtatggct agcatgactg gtggacagca aatgggtcgg
     2881 gatctgtacg acgatgacga taaggatccg agcgtgatca agacggatat gaaaatccgc
     2941 ctccagatgc agggggccgt caacggacac cctttcgtta ttaccgggga gggcgagggc
     3001 aaaccctatg aaggcaaaca cacaatcaac cttacagtgc aggacggcgg tccgctcccc
     3061 ttcgcctttg atatcctgac cacagctttc caatacggga atagggtgtt caccaagtac
     3121 cctaaggaca ttcccgatta ctttaagcag agctttcctg ccggatattc ctgggagcgg
     3181 tgcatgacct tcgaagacgg tggcctgtgc accgtgagtt cacatatcaa gattgaggga
     3241 gattatttca cctacgatat caggtttcac ggagtcaatt tccccgccgg cggtcccgtc
     3301 atgcagaaaa agactctgcg gtgggaacca agcaccgaga acatgtacgt gcgagacggt
     3361 gtgctggtag gcgaggtgga gagaactctg ctgctggagg gcaataaaca tcaccgctgt
     3421 aacttcagaa caacttacaa agcaaagaaa gaagtagtcc tgccagaata ccactttgtt
     3481 gaccaccgga tagagatctt ggggcatgac aaggactata acaatgtggt tgtgtatgaa
     3541 aacgcagtgg ctcggcaaca ggcctctacc ctgccaagca aggcgaagta gga
//
