LOCUS       sequence-200344-        4391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding moxGFP.
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-moxGFP
FEATURES             Location/Qualifiers
     source          1..4391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             201..914
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FISTTGKLPVPWPTLVTTLTYGVQSFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(231..259)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     854..874
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      955..1041
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1579..2529)
                     /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      2577..2799
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3291..3385)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3416..4210)
                     /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(3527..3546)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4137..4156
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     join(4389..4391,1..16)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aggcgtatca cgaggccctt tcgtcttcac cacagctaac accacgtcgt ccctatctgc
       61 tgccctaggt ctatgagtgg ttgctggata actttacggg catgcataag gctcgtatga
      121 tatattcagg gagaccacaa cggtttccct ctacaaataa ttttgtttaa cttttctaga
      181 tttaagaagg agatatacat atgagtaaag gagaagaact tttcactgga gttgtcccaa
      241 ttcttgttga attagatggt gatgttaatg ggcacaaatt ttctgtccgt ggagagggtg
      301 aaggtgatgc aacaaacgga aaacttaccc ttaaatttat ttccactact ggaaaactac
      361 ctgttccatg gccaacactt gtcactactt taacttatgg tgttcaatcg ttttcaagat
      421 acccagatca tatgaaacag catgactttt tcaagagtgc catgcccgaa ggttatgtac
      481 aggaaagaac tatatctttc aaagatgacg ggacctacaa gacacgtgct gaagtcaagt
      541 ttgaaggtga tacccttgtt aatagaatcg agttaaaagg tattgatttt aaagaagatg
      601 gaaacattct tggacacaaa ttggaataca actttaactc acacaatgta tacatcacgg
      661 cagacaaaca aaagaatgga atcaaagcta acttcaaaat tagacacaac gttgaagatg
      721 gaagcgttca actagcagac cattatcaac aaaatactcc aattggcgat ggccctgtcc
      781 ttttaccaga caaccattac ctgtccacac aatctaagct ttcgaaagat cccaacgaaa
      841 agagagacca catggtcctt cttgagtttg taacagctgc tgggattaca catggcatgg
      901 atgaactata caaataaatg tccagacctg caggcatgca agctctagag gcatcaaata
      961 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     1021 gctctcctga gtaggacaaa tccgccgccc tagacctagg gtacgggttt tgctgcccgc
     1081 aaacgggctg ttctggtgtt gctagtttgt tatcagaatc gcagatccgg cttcagccgg
     1141 tttgccggct gaaagcgcta tttcttccag aattgccatg attttttccc cacgggaggc
     1201 gtcactggct cccgtgttgt cggcagcttt gattcgataa gcagcatcgc ctgtttcagg
     1261 ctgtctatgt gtgactgttg agctgtaaca agttgtctca ggtgttcaat ttcatgttct
     1321 agttgctttg ttttactggt ttcacctgtt ctattaggtg ttacatgctg ttcatctgtt
     1381 acattgtcga tctgttcatg gtgaacagct ttgaatgcac caaaaactcg taaaagctct
     1441 gatgtatcta tcttttttac accgttttca tctgtgcata tggacagttt tccctttgat
     1501 atgtaacggt gaacagttgt tctacttttg tttgttagtc ttgatgcttc actgatagat
     1561 acaagagcca taagaacctc agatccttcc gtatttagcc agtatgttct ctagtgtggt
     1621 tcgttgtttt tgcgtgagcc atgagaacga accattgaga tcatacttac tttgcatgtc
     1681 actcaaaaat tttgcctcaa aactggtgag ctgaattttt gcagttaaag catcgtgtag
     1741 tgtttttctt agtccgttat gtaggtagga atctgatgta atggttgttg gtattttgtc
     1801 accattcatt tttatctggt tgttctcaag ttcggttacg agatccattt gtctatctag
     1861 ttcaacttgg aaaatcaacg tatcagtcgg gcggcctcgc ttatcaacca ccaatttcat
     1921 attgctgtaa gtgtttaaat ctttacttat tggtttcaaa acccattggt taagcctttt
     1981 aaactcatgg tagttatttt caagcattaa catgaactta aattcatcaa ggctaatctc
     2041 tatatttgcc ttgtgagttt tcttttgtgt tagttctttt aataaccact cataaatcct
     2101 catagagtat ttgttttcaa aagacttaac atgttccaga ttatatttta tgaatttttt
     2161 taactggaaa agataaggca atatctcttc actaaaaact aattctaatt tttcgcttga
     2221 gaacttggca tagtttgtcc actggaaaat ctcaaagcct ttaaccaaag gattcctgat
     2281 ttccacagtt ctcgtcatca gctctctggt tgctttagct aatacaccat aagcattttc
     2341 cctactgatg ttcatcatct gagcgtattg gttataagtg aacgataccg tccgttcttt
     2401 ccttgtaggg ttttcaatcg tggggttgag tagtgccaca cagcataaaa ttagcttggt
     2461 ttcatgctcc gttaagtcat agcgactaat cgctagttca tttgctttga aaacaactaa
     2521 ttcagacata catctcaatt ggtctaggtg attttaatca ctataccaat tgagatgggc
     2581 tagtcaatga taattactag tccttttcct ttgagttgtg ggtatctgta aattctgcta
     2641 gacctttgct ggaaaacttg taaattctgc tagaccctct gtaaattccg ctagaccttt
     2701 gtgtgttttt tttgtttata ttcaagtggt tataatttat agaataaaga aagaataaaa
     2761 aaagataaaa agaatagatc ccagccctgt gtataactca ctactttagt cagttccgca
     2821 gtattacaaa aggatgtcgc aaacgctgtt tgctcctcta caaaacagac cttaaaaccc
     2881 taaaggctta agtagcaccc tcgcaagctc gggcaaatcg ctgaatattc cttttgtctc
     2941 cgaccatcag gcacctgagt cgctgtcttt ttcgtgacat tcagttcgct gcgctcacgg
     3001 ctctggcagt gaatgggggt aaatggcact acaggcgcct tttatggatt catgcaagga
     3061 aactacccat aatacaagaa aagcccgtca cgggcttctc agggcgtttt atggcgggtc
     3121 tgctatgtgg tgctatctga ctttttgctg ttcagcagtt cctgccctct gattttccag
     3181 tctgaccact tcggattatc ccgtgacagg tcattcagac tggctaatgc acccagtaag
     3241 gcagcggtat catcaacagg cttacccgtc ttactgtccc tagtgcttgg attctcacca
     3301 ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc
     3361 attactggat ctatcaacag gagtccaagc gagctctcga accccagagt cccgctcaga
     3421 agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt
     3481 aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag
     3541 ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag
     3601 aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga
     3661 gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct ggcgcgagcc
     3721 cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg
     3781 ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat
     3841 gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg
     3901 acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga
     3961 caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg
     4021 cctcgtcctg cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc
     4081 gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc
     4141 agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt
     4201 gttcaatcat gcgaaacgat cctcatcctg tctcttgatc agatcttgat cccctgcgcc
     4261 atcagatcct tggcggcaag aaagccatcc agtttacttt gcagggcttc ccaaccttac
     4321 cagagggcgc cccagctggc aattccgacg tctaagaaac cattattatc atgacattaa
     4381 cctataaaaa t
//
