LOCUS       sequence-200322-        4391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding sfGFP.
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-sfGFP
FEATURES             Location/Qualifiers
     source          1..4391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             182..895
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(212..240)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     835..855
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      936..1022
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1560..2510)
                     /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      2558..2780
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3272..3366)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3397..4191)
                     /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(3508..3527)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4118..4137
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4370..4388
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttcgtcttca ccacagctaa caccacgtcg tccctatctg ctgccctagg tctatgagtg
       61 gttgctggat aactttacgg gcatgcataa ggctcgtatg atatattcag ggagaccaca
      121 acggtttccc tctacaaata attttgttta acttttctag atttaagaag gagatataca
      181 tatgagtaaa ggagaagaac ttttcactgg agttgtccca attcttgttg aattagatgg
      241 tgatgttaat gggcacaaat tttctgtccg tggagagggt gaaggtgatg caacaaacgg
      301 aaaacttacc cttaaattta tttgcactac tggaaaacta cctgttccat ggccaacact
      361 tgtcactact ttaacttatg gtgttcaatg cttttcaaga tacccagatc atatgaaaca
      421 gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaaagaa ctatatcttt
      481 caaagatgac gggacctaca agacacgtgc tgaagtcaag tttgaaggtg atacccttgt
      541 taatagaatc gagttaaaag gtattgattt taaagaagat ggaaacattc ttggacacaa
      601 attggaatac aactttaact cacacaatgt atacatcacg gcagacaaac aaaagaatgg
      661 aatcaaagct aacttcaaaa ttagacacaa cgttgaagat ggaagcgttc aactagcaga
      721 ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta
      781 cctgtccaca caatctgtcc tttcgaaaga tcccaacgaa aagagagacc acatggtcct
      841 tcttgagttt gtaacagctg ctgggattac acatggcatg gatgaactat acaaataaat
      901 gtccagacct gcaggcatgc aagctctaga ggcatcaaat aaaacgaaag gctcagtcga
      961 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     1021 atccgccgcc ctagacctag ggtacgggtt ttgctgcccg caaacgggct gttctggtgt
     1081 tgctagtttg ttatcagaat cgcagatccg gcttcagccg gtttgccggc tgaaagcgct
     1141 atttcttcca gaattgccat gattttttcc ccacgggagg cgtcactggc tcccgtgttg
     1201 tcggcagctt tgattcgata agcagcatcg cctgtttcag gctgtctatg tgtgactgtt
     1261 gagctgtaac aagttgtctc aggtgttcaa tttcatgttc tagttgcttt gttttactgg
     1321 tttcacctgt tctattaggt gttacatgct gttcatctgt tacattgtcg atctgttcat
     1381 ggtgaacagc tttgaatgca ccaaaaactc gtaaaagctc tgatgtatct atctttttta
     1441 caccgttttc atctgtgcat atggacagtt ttccctttga tatgtaacgg tgaacagttg
     1501 ttctactttt gtttgttagt cttgatgctt cactgataga tacaagagcc ataagaacct
     1561 cagatccttc cgtatttagc cagtatgttc tctagtgtgg ttcgttgttt ttgcgtgagc
     1621 catgagaacg aaccattgag atcatactta ctttgcatgt cactcaaaaa ttttgcctca
     1681 aaactggtga gctgaatttt tgcagttaaa gcatcgtgta gtgtttttct tagtccgtta
     1741 tgtaggtagg aatctgatgt aatggttgtt ggtattttgt caccattcat ttttatctgg
     1801 ttgttctcaa gttcggttac gagatccatt tgtctatcta gttcaacttg gaaaatcaac
     1861 gtatcagtcg ggcggcctcg cttatcaacc accaatttca tattgctgta agtgtttaaa
     1921 tctttactta ttggtttcaa aacccattgg ttaagccttt taaactcatg gtagttattt
     1981 tcaagcatta acatgaactt aaattcatca aggctaatct ctatatttgc cttgtgagtt
     2041 ttcttttgtg ttagttcttt taataaccac tcataaatcc tcatagagta tttgttttca
     2101 aaagacttaa catgttccag attatatttt atgaattttt ttaactggaa aagataaggc
     2161 aatatctctt cactaaaaac taattctaat ttttcgcttg agaacttggc atagtttgtc
     2221 cactggaaaa tctcaaagcc tttaaccaaa ggattcctga tttccacagt tctcgtcatc
     2281 agctctctgg ttgctttagc taatacacca taagcatttt ccctactgat gttcatcatc
     2341 tgagcgtatt ggttataagt gaacgatacc gtccgttctt tccttgtagg gttttcaatc
     2401 gtggggttga gtagtgccac acagcataaa attagcttgg tttcatgctc cgttaagtca
     2461 tagcgactaa tcgctagttc atttgctttg aaaacaacta attcagacat acatctcaat
     2521 tggtctaggt gattttaatc actataccaa ttgagatggg ctagtcaatg ataattacta
     2581 gtccttttcc tttgagttgt gggtatctgt aaattctgct agacctttgc tggaaaactt
     2641 gtaaattctg ctagaccctc tgtaaattcc gctagacctt tgtgtgtttt ttttgtttat
     2701 attcaagtgg ttataattta tagaataaag aaagaataaa aaaagataaa aagaatagat
     2761 cccagccctg tgtataactc actactttag tcagttccgc agtattacaa aaggatgtcg
     2821 caaacgctgt ttgctcctct acaaaacaga ccttaaaacc ctaaaggctt aagtagcacc
     2881 ctcgcaagct cgggcaaatc gctgaatatt ccttttgtct ccgaccatca ggcacctgag
     2941 tcgctgtctt tttcgtgaca ttcagttcgc tgcgctcacg gctctggcag tgaatggggg
     3001 taaatggcac tacaggcgcc ttttatggat tcatgcaagg aaactaccca taatacaaga
     3061 aaagcccgtc acgggcttct cagggcgttt tatggcgggt ctgctatgtg gtgctatctg
     3121 actttttgct gttcagcagt tcctgccctc tgattttcca gtctgaccac ttcggattat
     3181 cccgtgacag gtcattcaga ctggctaatg cacccagtaa ggcagcggta tcatcaacag
     3241 gcttacccgt cttactgtcc ctagtgcttg gattctcacc aataaaaaac gcccggcggc
     3301 aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca
     3361 ggagtccaag cgagctctcg aaccccagag tcccgctcag aagaactcgt caagaaggcg
     3421 atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc
     3481 agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata
     3541 gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac
     3601 catgatattc ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat
     3661 gcgcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag
     3721 atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt
     3781 cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc
     3841 agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg
     3901 cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc
     3961 gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt
     4021 cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg
     4081 gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct
     4141 ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga
     4201 tcctcatcct gtctcttgat cagatcttga tcccctgcgc catcagatcc ttggcggcaa
     4261 gaaagccatc cagtttactt tgcagggctt cccaacctta ccagagggcg ccccagctgg
     4321 caattccgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     4381 cacgaggccc t
//
