LOCUS       sequence-199042-        3599 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the fluorescent protein efasCFP for constitutive expression 
            in E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3599)
  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 3599)
  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: pRSM14
FEATURES             Location/Qualifiers
     source          1..3599
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(62..80)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      76..123
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      220..675
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(307..326)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     517..538
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        701..805
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(806..874,875..1666)
                     /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(1024..1043)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1837..2425
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2326..2345
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2579..2596
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2681..2700
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2681..2699
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2783..2805
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2825..2842
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2846..2878
                     /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(2882..2890,2891..2905)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
ORIGIN
        1 aattcgaagc ttgatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
       61 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
      121 ttgctgaaag gaggaactat atccggatct ggcgtaatag cgaagaggcc agcaccgatc
      181 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
      241 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
      301 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
      361 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
      421 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
      481 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
      541 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
      601 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
      661 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
      721 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
      781 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
      841 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
      901 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
      961 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     1021 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     1081 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     1141 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     1201 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     1261 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     1321 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     1381 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     1441 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     1501 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     1561 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     1621 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     1681 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     1741 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     1801 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     1861 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     1921 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     1981 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2041 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2101 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2161 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     2221 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     2281 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     2341 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     2401 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     2461 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     2521 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     2581 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     2641 ttggccgatt cattaatgca ggatctcgat cccgcgaaat taatacgact cactataggg
     2701 agaccacaac ggtttccctc tagaatcacc gagtttattc ttgacacctg atgcgatgaa
     2761 tgatataata ggaaagtact gttttgttta actttaagaa ggagatatac atatgcgggg
     2821 ttctcatcat catcatcatc atggtatggc tagcatgact ggtggacagc aaatgggtcg
     2881 ggatctgtac gacgatgacg ataaggatcc gagcttctct aagcaggtgc tgaatgatgt
     2941 tactatgacc tacttcatgg aaggtagcgt gaacgggcac gactttacaa tcgaggggga
     3001 gggcacggga aaaccctatg agggccacca acggtttaac ctgcgtgtga ccaaaggcgc
     3061 cccactgcct ttcgccgtgg acatcttgtc agcagccttt gcttatggta acaggtgctt
     3121 cacaaagtac cccaaggaga ttcccgattt ttttaagcag tcacttccgg aggacatgag
     3181 ctgggaaagg accatgacct ttgaggatgg cggaatcgtg gcgattagtg cccacattag
     3241 actgataggc aatcggttcg agcataaaag caagttcgta ggggtgaact ttccagctga
     3301 cggacctgtc atgcaacgga agaccctcgg ctgggaaccc tcctctgaaa agatgacccc
     3361 acgcgatggc atcctgaaag gatacgtgcc cagcttcctg gtcctgcagg gcggtggaaa
     3421 ctacagatgc gattacgata ctagttatcg ggccatgaag cccgtcgaga tgcctggggg
     3481 gcacttcatc cagcaccgaa ttgttcggcg cgacatcaaa aaggacagca atggcaacac
     3541 atggcagatc caggaggacg ccttcgcaca taattccgaa gtgcccgact ccgcttagg
//
