LOCUS       sequence-195172-        3575 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the fluorescent protein KCyG4219 for constitutive expression
            in E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3575)
  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 3575)
  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: pRSM4
FEATURES             Location/Qualifiers
     source          1..3575
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(63..81)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      77..124
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      221..676
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(308..327)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     518..539
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        702..806
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(807..875,876..1667)
                     /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(1025..1044)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1838..2426
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2327..2346
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2580..2597
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2682..2701
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2682..2700
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2784..2806
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2826..2843
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2847..2879
                     /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(2883..2891,2892..2906)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
ORIGIN
        1 gaattcgaag cttgatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
       61 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
      121 tttgctgaaa ggaggaacta tatccggatc tggcgtaata gcgaagaggc cagcaccgat
      181 cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca
      241 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
      301 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
      361 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
      421 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
      481 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
      541 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
      601 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
      661 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
      721 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
      781 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
      841 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
      901 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
      961 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     1021 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     1081 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     1141 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     1201 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     1261 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     1321 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     1381 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     1441 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     1501 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     1561 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     1621 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     1681 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     1741 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     1801 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1861 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1921 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1981 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     2041 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     2101 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2161 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2221 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2281 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2341 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2401 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     2461 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     2521 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     2581 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     2641 gttggccgat tcattaatgc aggatctcga tcccgcgaaa ttaatacgac tcactatagg
     2701 gagaccacaa cggtttccct ctagaatcac cgagtttatt cttgacacct gatgcgatga
     2761 atgatataat aggaaagtac tgttttgttt aactttaaga aggagatata catatgcggg
     2821 gttctcatca tcatcatcat catggtatgg ctagcatgac tggtggacag caaatgggtc
     2881 gggatctgta cgacgatgac gataaggatc cggtgagcgt gatcaagcca gaaatgaaga
     2941 tgcggtacta tatggacggc agcgtcaacg ggcatgagtt caccgtggag ggggaaggca
     3001 ccggccggcc atacgaaggt aaacacaaga tcactctgga cgtgaccaag ggcggacctc
     3061 tgccctttgc gttcgatctg ctctcaactg ttttcagcta cgggaacaga gccctgacca
     3121 aataccccga tgatattccc gactacttta aacagtgctt cccggggggg tatagctggg
     3181 agaggaagtt cgagttcgag gacggcggac tcgccatcgc taaagcagaa atctccctga
     3241 agggaaattg ttttgaacac aaatctacca ttgagggtac gttccccgat tctagtccta
     3301 tcatgcagaa taaaaccctg ggttgggagc ctagcacaga aaagatgaca gtgcgcgacg
     3361 gctcaatgaa gggcgacgat gccgcttatc tgaagcttgt ggggggagga aaccacaagt
     3421 gctactttac aacaacttat accgccaaaa agaagattcc caacctgccc ggcagccact
     3481 ttatcggtca ccggatttcc tccgtggtcg agggcaccaa aatcaaggta atggaggacg
     3541 ccatagcaca tttgtaccca ttcaatggaa gttag
//
