LOCUS       sequence-267561-        4109 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes a photostable monomeric superfolder GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4109)
  AUTHORS   Valbuena FM, Fizgerald I, Strack RL, Andruska N, Smith L, Glick BS
  TITLE     A photostable monomeric superfolder GFP.
  JOURNAL   Traffic. 2020 May 15. doi: 10.1111/tra.12737.
  PUBMED    32415747
REFERENCE   2  (bases 1 to 4109)
  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: "Traffic. 
            2020 May 15. doi: 10.1111/tra.12737."
COMMENT     Sequence Label: pQE-60NA-msGFP2
FEATURES             Location/Qualifiers
     source          1..4109
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     156..174
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(198..212,213..218,219..235,236..242)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    218..234
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     protein_bind    250..266
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             285..296
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             303..1010
                     /codon_start=1
                     /product="photostable monomeric superfolder GFP (Valbuena 
                     et al., 2020)"
                     /label=msGFP2
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTITFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITGGSGGS"
     primer_bind     complement(345..366)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     953..974
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      1044..1138
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1248..1267)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1910..1996
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2043..2065)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2151..2291
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2206..2225
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2306..2323)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2477..3065)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2557..2576)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3236..4027,4028..4096))
                     /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     3859..3878
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4097..4109,1..92))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
       61 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgct
      121 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
      181 gtcttcacct cgagaaatca taaaaaattt atttgctttg tgagcggata acaattataa
      241 tagattcaat tgtgagcgga taacaatttc acacagaatt cattaaagag gagaaattaa
      301 ccatggatag cactgagagc ctgttcaccg gggtggtgcc catcctggtc gagctggacg
      361 gcgacgtaaa cggccacaag ttcagcgtgc gcggcgaggg cgagggcgat gccaccaacg
      421 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
      481 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
      541 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcacct
      601 tcaaggacga cggcacctac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
      661 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
      721 agctggagta caactacaac agccacaacg tctatatcac cgccgacaag cagaagaacg
      781 gcatcaaggc caacttcaag atccgccaca acgtggagga cggcagcgtg cagctcgccg
      841 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
      901 acctgagcac ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc
      961 tgctggagtt cgtgaccgcc gccgggatca ctggaggcag tggcggttct taaagcggcc
     1021 gcaagcttaa ttagctgagc ttggactcct gttgatagat ccagtaatga cctcagaact
     1081 ccatctggat ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtgagaatcc
     1141 aagctagctt ggcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg
     1201 gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt
     1261 cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga
     1321 ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga
     1381 tgaatgctca tccggaattt cgtatggcaa tgaaagacgg tgagctggtg atatgggata
     1441 gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga
     1501 gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt
     1561 acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtctcag
     1621 ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct
     1681 tcgcccccgt tttcaccatg catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
     1741 ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt
     1801 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt
     1861 tattggtgcc cttaaacgcc tggggtaatg actctctagc ttgaggcatc aaataaaacg
     1921 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     1981 cctgagtagg acaaatccgc cctctagagc tgcctcgcgc gtttcggtga tgacggtgaa
     2041 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     2101 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg
     2161 acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga
     2221 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     2281 accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2341 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2401 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2461 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2521 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2581 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2641 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2701 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2761 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2821 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2881 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     2941 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3001 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3061 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3121 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3181 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3241 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3301 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3361 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3421 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3481 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     3541 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     3601 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     3661 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     3721 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     3781 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     3841 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     3901 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     3961 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4021 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4081 aatgttgaat actcatactc ttccttttt
//
