LOCUS       sequence-200292-        4391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding SCFP1.
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-SCFP1
FEATURES             Location/Qualifiers
     source          1..4391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             154..867
                     /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
                     FICTTGKLPVPWPTLVTTLTWGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(184..212)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     807..827
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      908..994
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1532..2482)
                     /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      2530..2752
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3244..3338)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3369..4163)
                     /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(3480..3499)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4090..4109
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4342..4360
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cgtccctatc tgctgcccta ggtctatgag tggttgctgg ataactttac gggcatgcat
       61 aaggctcgta tgatatattc agggagacca caacggtttc cctctacaaa taattttgtt
      121 taacttttct agatttaaga aggagatata catatgagta aaggagaaga acttttcact
      181 ggagttgtcc caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc
      241 agtggagagg gtgaaggtga tgcaacatac ggaaaactta cccttaaatt tatttgcact
      301 actggaaaac tacctgttcc atggccaaca cttgtcacta ctctcacttg gggtcttcaa
      361 tgctttgcaa gatacccaga tcatatgaaa cagcatgact ttttcaagag tgccatgccc
      421 gaaggttatg tacaggaaag aactatattt ttcaaagatg acgggaacta caagacacgt
      481 gctgaagtca agtttgaagg tgataccctt gttaatagaa tcgagttaaa aggtattgat
      541 tttaaagaag atggaaacat tcttggacac aaattggaat acaactacat ctcacacaat
      601 gtatacatca cggcagacaa acaaaagaat ggaatcaaag ctaacttcaa aattagacac
      661 aacattgaag atggaggcgt tcaactagca gaccattatc aacaaaatac tccaattggc
      721 gatggccctg tccttttacc agacaaccat tacctgtcca cacaatctaa gctttcgaaa
      781 gatcccaacg aaaagagaga ccacatggtc cttcttgagt ttgtaacagc tgctgggatt
      841 acacatggca tggatgaact atacaaataa atgtccagac ctgcaggcat gcaagctcta
      901 gaggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
      961 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ccctagacct agggtacggg
     1021 ttttgctgcc cgcaaacggg ctgttctggt gttgctagtt tgttatcaga atcgcagatc
     1081 cggcttcagc cggtttgccg gctgaaagcg ctatttcttc cagaattgcc atgatttttt
     1141 ccccacggga ggcgtcactg gctcccgtgt tgtcggcagc tttgattcga taagcagcat
     1201 cgcctgtttc aggctgtcta tgtgtgactg ttgagctgta acaagttgtc tcaggtgttc
     1261 aatttcatgt tctagttgct ttgttttact ggtttcacct gttctattag gtgttacatg
     1321 ctgttcatct gttacattgt cgatctgttc atggtgaaca gctttgaatg caccaaaaac
     1381 tcgtaaaagc tctgatgtat ctatcttttt tacaccgttt tcatctgtgc atatggacag
     1441 ttttcccttt gatatgtaac ggtgaacagt tgttctactt ttgtttgtta gtcttgatgc
     1501 ttcactgata gatacaagag ccataagaac ctcagatcct tccgtattta gccagtatgt
     1561 tctctagtgt ggttcgttgt ttttgcgtga gccatgagaa cgaaccattg agatcatact
     1621 tactttgcat gtcactcaaa aattttgcct caaaactggt gagctgaatt tttgcagtta
     1681 aagcatcgtg tagtgttttt cttagtccgt tatgtaggta ggaatctgat gtaatggttg
     1741 ttggtatttt gtcaccattc atttttatct ggttgttctc aagttcggtt acgagatcca
     1801 tttgtctatc tagttcaact tggaaaatca acgtatcagt cgggcggcct cgcttatcaa
     1861 ccaccaattt catattgctg taagtgttta aatctttact tattggtttc aaaacccatt
     1921 ggttaagcct tttaaactca tggtagttat tttcaagcat taacatgaac ttaaattcat
     1981 caaggctaat ctctatattt gccttgtgag ttttcttttg tgttagttct tttaataacc
     2041 actcataaat cctcatagag tatttgtttt caaaagactt aacatgttcc agattatatt
     2101 ttatgaattt ttttaactgg aaaagataag gcaatatctc ttcactaaaa actaattcta
     2161 atttttcgct tgagaacttg gcatagtttg tccactggaa aatctcaaag cctttaacca
     2221 aaggattcct gatttccaca gttctcgtca tcagctctct ggttgcttta gctaatacac
     2281 cataagcatt ttccctactg atgttcatca tctgagcgta ttggttataa gtgaacgata
     2341 ccgtccgttc tttccttgta gggttttcaa tcgtggggtt gagtagtgcc acacagcata
     2401 aaattagctt ggtttcatgc tccgttaagt catagcgact aatcgctagt tcatttgctt
     2461 tgaaaacaac taattcagac atacatctca attggtctag gtgattttaa tcactatacc
     2521 aattgagatg ggctagtcaa tgataattac tagtcctttt cctttgagtt gtgggtatct
     2581 gtaaattctg ctagaccttt gctggaaaac ttgtaaattc tgctagaccc tctgtaaatt
     2641 ccgctagacc tttgtgtgtt ttttttgttt atattcaagt ggttataatt tatagaataa
     2701 agaaagaata aaaaaagata aaaagaatag atcccagccc tgtgtataac tcactacttt
     2761 agtcagttcc gcagtattac aaaaggatgt cgcaaacgct gtttgctcct ctacaaaaca
     2821 gaccttaaaa ccctaaaggc ttaagtagca ccctcgcaag ctcgggcaaa tcgctgaata
     2881 ttccttttgt ctccgaccat caggcacctg agtcgctgtc tttttcgtga cattcagttc
     2941 gctgcgctca cggctctggc agtgaatggg ggtaaatggc actacaggcg ccttttatgg
     3001 attcatgcaa ggaaactacc cataatacaa gaaaagcccg tcacgggctt ctcagggcgt
     3061 tttatggcgg gtctgctatg tggtgctatc tgactttttg ctgttcagca gttcctgccc
     3121 tctgattttc cagtctgacc acttcggatt atcccgtgac aggtcattca gactggctaa
     3181 tgcacccagt aaggcagcgg tatcatcaac aggcttaccc gtcttactgt ccctagtgct
     3241 tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg
     3301 gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctct cgaaccccag
     3361 agtcccgctc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga
     3421 gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca
     3481 atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag
     3541 tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca
     3601 tgggtcacga cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc gaacagttcg
     3661 gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc
     3721 atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc
     3781 ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga
     3841 gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt
     3901 cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac
     3961 gatagccgcg ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc ggtcttgaca
     4021 aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt
     4081 gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg
     4141 tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcagatctt
     4201 gatcccctgc gccatcagat ccttggcggc aagaaagcca tccagtttac tttgcagggc
     4261 ttcccaacct taccagaggg cgccccagct ggcaattccg acgtctaaga aaccattatt
     4321 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct tcaccacagc
     4381 taacaccacg t
//
