LOCUS       sequence-200473-        4391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding SCFP3A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4391)
  AUTHORS   Balleza E, Kim JM, Cluzel P
  TITLE     Systematic characterization of maturation time of fluorescent 
            proteins in living cells.
  JOURNAL   Nat Methods. 2018 Jan;15(1):47-51. doi: 10.1038/nmeth.4509. Epub 
            2017 Nov 20.
  PUBMED    29320486
REFERENCE   2  (bases 1 to 4391)
  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: 
            "10.1038/nmeth.4509"; journalName: "Nat Methods"; date: "2018-01"; 
            volume: "15"; issue: "1"; pages: "47-51"
COMMENT     Sequence Label: pEB1-SCFP3A
FEATURES             Location/Qualifiers
     source          1..4391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     56..74
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             259..972
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISDNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(289..317)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     912..932
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1013..1099
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1637..2587)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2635..2857
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3349..3443)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3474..4268)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3585..3604)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4195..4214
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 agctggcaat tccgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
       61 gcgtatcacg aggccctttc gtcttcacca cagctaacac cacgtcgtcc ctatctgctg
      121 ccctaggtct atgagtggtt gctggataac tttacgggca tgcataaggc tcgtatgata
      181 tattcaggga gaccacaacg gtttccctct acaaataatt ttgtttaact tttctagatt
      241 taagaaggag atatacatat gagtaaagga gaagaacttt tcactggagt tgtcccaatt
      301 cttgttgaat tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa
      361 ggtgatgcaa catacggaaa acttaccctt aaatttattt gcactactgg aaaactacct
      421 gttccatggc caacacttgt cactactctc acttggggtg ttcaatgctt tgcaagatac
      481 ccagatcata tgaaacagca tgactttttc aagagtgcca tgcccgaagg ttatgtacag
      541 gaaagaacta tatttttcaa agatgacggg aactacaaga cacgtgctga agtcaagttt
      601 gaaggtgata cccttgttaa tagaatcgag ttaaaaggta ttgattttaa agaagatgga
      661 aacattcttg gacacaaatt ggaatacaac tacatctcag acaatgtata catcacggca
      721 gacaaacaaa agaatggaat caaagctaac ttcaaaatta gacacaacat tgaagatgga
      781 ggcgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt
      841 ttaccagaca accattacct gtccacacaa tctaagcttt cgaaagatcc caacgaaaag
      901 agagaccaca tggtccttct tgagtttgta acagctgctg ggattacaca tggcatggat
      961 gaactataca aataaatgtc cagacctgca ggcatgcaag ctctagaggc atcaaataaa
     1021 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     1081 tctcctgagt aggacaaatc cgccgcccta gacctagggt acgggttttg ctgcccgcaa
     1141 acgggctgtt ctggtgttgc tagtttgtta tcagaatcgc agatccggct tcagccggtt
     1201 tgccggctga aagcgctatt tcttccagaa ttgccatgat tttttcccca cgggaggcgt
     1261 cactggctcc cgtgttgtcg gcagctttga ttcgataagc agcatcgcct gtttcaggct
     1321 gtctatgtgt gactgttgag ctgtaacaag ttgtctcagg tgttcaattt catgttctag
     1381 ttgctttgtt ttactggttt cacctgttct attaggtgtt acatgctgtt catctgttac
     1441 attgtcgatc tgttcatggt gaacagcttt gaatgcacca aaaactcgta aaagctctga
     1501 tgtatctatc ttttttacac cgttttcatc tgtgcatatg gacagttttc cctttgatat
     1561 gtaacggtga acagttgttc tacttttgtt tgttagtctt gatgcttcac tgatagatac
     1621 aagagccata agaacctcag atccttccgt atttagccag tatgttctct agtgtggttc
     1681 gttgtttttg cgtgagccat gagaacgaac cattgagatc atacttactt tgcatgtcac
     1741 tcaaaaattt tgcctcaaaa ctggtgagct gaatttttgc agttaaagca tcgtgtagtg
     1801 tttttcttag tccgttatgt aggtaggaat ctgatgtaat ggttgttggt attttgtcac
     1861 cattcatttt tatctggttg ttctcaagtt cggttacgag atccatttgt ctatctagtt
     1921 caacttggaa aatcaacgta tcagtcgggc ggcctcgctt atcaaccacc aatttcatat
     1981 tgctgtaagt gtttaaatct ttacttattg gtttcaaaac ccattggtta agccttttaa
     2041 actcatggta gttattttca agcattaaca tgaacttaaa ttcatcaagg ctaatctcta
     2101 tatttgcctt gtgagttttc ttttgtgtta gttcttttaa taaccactca taaatcctca
     2161 tagagtattt gttttcaaaa gacttaacat gttccagatt atattttatg aattttttta
     2221 actggaaaag ataaggcaat atctcttcac taaaaactaa ttctaatttt tcgcttgaga
     2281 acttggcata gtttgtccac tggaaaatct caaagccttt aaccaaagga ttcctgattt
     2341 ccacagttct cgtcatcagc tctctggttg ctttagctaa tacaccataa gcattttccc
     2401 tactgatgtt catcatctga gcgtattggt tataagtgaa cgataccgtc cgttctttcc
     2461 ttgtagggtt ttcaatcgtg gggttgagta gtgccacaca gcataaaatt agcttggttt
     2521 catgctccgt taagtcatag cgactaatcg ctagttcatt tgctttgaaa acaactaatt
     2581 cagacataca tctcaattgg tctaggtgat tttaatcact ataccaattg agatgggcta
     2641 gtcaatgata attactagtc cttttccttt gagttgtggg tatctgtaaa ttctgctaga
     2701 cctttgctgg aaaacttgta aattctgcta gaccctctgt aaattccgct agacctttgt
     2761 gtgttttttt tgtttatatt caagtggtta taatttatag aataaagaaa gaataaaaaa
     2821 agataaaaag aatagatccc agccctgtgt ataactcact actttagtca gttccgcagt
     2881 attacaaaag gatgtcgcaa acgctgtttg ctcctctaca aaacagacct taaaacccta
     2941 aaggcttaag tagcaccctc gcaagctcgg gcaaatcgct gaatattcct tttgtctccg
     3001 accatcaggc acctgagtcg ctgtcttttt cgtgacattc agttcgctgc gctcacggct
     3061 ctggcagtga atgggggtaa atggcactac aggcgccttt tatggattca tgcaaggaaa
     3121 ctacccataa tacaagaaaa gcccgtcacg ggcttctcag ggcgttttat ggcgggtctg
     3181 ctatgtggtg ctatctgact ttttgctgtt cagcagttcc tgccctctga ttttccagtc
     3241 tgaccacttc ggattatccc gtgacaggtc attcagactg gctaatgcac ccagtaaggc
     3301 agcggtatca tcaacaggct tacccgtctt actgtcccta gtgcttggat tctcaccaat
     3361 aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt ctgaggtcat
     3421 tactggatct atcaacagga gtccaagcga gctctcgaac cccagagtcc cgctcagaag
     3481 aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa
     3541 agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc
     3601 aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa
     3661 aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga
     3721 tcctcgccgt cgggcatgcg cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc
     3781 tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct
     3841 cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc
     3901 agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac
     3961 aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca
     4021 acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc
     4081 tcgtcctgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc
     4141 ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag
     4201 tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt
     4261 tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcag atcttgatcc cctgcgccat
     4321 cagatccttg gcggcaagaa agccatccag tttactttgc agggcttccc aaccttacca
     4381 gagggcgccc c
//
