LOCUS       sequence-200541-        3968 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding 2ndVal mNeptune2.5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3968)
  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 3968)
  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: pEB2-2ndVal mNept2.5
FEATURES             Location/Qualifiers
     source          1..3968
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     66..84
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      1041..1087
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1260..2210)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2258..2480
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(2986..3020)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(3061..3855)
                     /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(3172..3191)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3782..3801
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 agggcgcccc agctggcaat tccgacgtct aagaaaccat tattatcatg acattaacct
       61 ataaaaatag gcgtatcacg aggccctttc gtcttcacca cagctaacac cacgtcgtcc
      121 ctatctgctg ccctaggtct atgagtggtt gctggataac tttacgggca tgcataaggc
      181 tcgtatgata tattcaggga gaccacaacg gtttccctct acaaataatt ttgtttaact
      241 tttctagatt taagaaggag atatacatat ggtgagtaaa ggagaagaac tgattaagga
      301 gaacatgcac accaagctgt acatggaggg caccgtgaac aaccaccact tcaagtgcac
      361 acacgagggc gaaggcaagc cctacgaggg cacccagacc aacagaatca aggtggtcga
      421 gggcggccct ttacccttcg ccttcgacat cctggctacc tgcttcatgt acggcagcaa
      481 gaccttcatc aaccacaccc agggcatccc cgacttcttt aagcagtcct tccctgaggg
      541 cttcacatgg gagagagtca ccacatacga agacgggggc gtgctgaccg ttacccagga
      601 caccagcctg caggacggct gcctgatcta caacgtcaag ctcagagggg tgaacttccc
      661 atccaacggc cctgtgatgc agaagaaaac actgggctgg gaggccagca ccgagaccct
      721 gtaccccgct gacggcggcc tggaaggcag atgcgacatg gccctgaagt tagtgggcgg
      781 gggccacctg cactgcaact tgaagaccac atacagatcc aagaaacccg ctaagaactt
      841 gaagatgccc ggcgtctact tcgtggacag gagactggaa agaatcaagg aggccgacaa
      901 tgagacctac gtcgagcagc acgaggtggc tgtggccagg tactgcgacc tgcctagcaa
      961 actggggcac aaacttaatg gcatggatga actatacaaa taaatgtcca gacctgcagg
     1021 catgcaagct ctagaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     1081 gttttatctg ttgaaatgca ccaaaaactc gtaaaagctc tgatgtatct atctttttta
     1141 caccgttttc atctgtgcat atggacagtt ttccctttga tatctaacgg tgaacagttg
     1201 ttctactttt gtttgttagt cttgatgctt cactgataga tacaagagcc ataagaacct
     1261 cagatccttc cgtatttagc cagtatgttc tctagtgtgg ttcgttgttt ttgcgtgagc
     1321 catgagaacg aaccattgag atcatgctta ctttgcatgt cactcaaaaa ttttgcctca
     1381 aaactggtga gctgaatttt tgcagttaaa gcatcgtgta gtgtttttct tagtccgtta
     1441 cgtaggtagg aatctgatgt aatggttgtt ggtattttgt caccattcat ttttatctgg
     1501 ttgttctcaa gttcggttac gagatccatt tgtctatcta gttcaacttg gaaaatcaac
     1561 gtatcagtcg ggcggcctcg cttatcaacc accaatttca tattgctgta agtgtttaaa
     1621 tctttactta ttggtttcaa aacccattgg ttaagccttt taaactcatg gtagttattt
     1681 tcaagcatta acatgaactt aaattcatca aggctaatct ctatatttgc cttgtgagtt
     1741 ttcttttgtg ttagttcttt taataaccac tcataaatcc tcatagagta tttgttttca
     1801 aaagacttaa catgttccag attatatttt atgaattttt ttaactggaa aagataaggc
     1861 aatatctctt cactaaaaac taattctaat ttttcgcttg agaacttggc atagtttgtc
     1921 cactggaaaa tctcaaagcc tttaaccaaa ggattcctga tttccacagt tctcgtcatc
     1981 agctctctgg ttgctttagc taatacacca taagcatttt ccctactgat gttcatcatc
     2041 tgagcgtatt ggttataagt gaacgatacc gtccgttctt tccttgtagg gttttcaatc
     2101 gtggggttga gtagtgccac acagcataaa attagcttgg tttcatgctc cgttaagtca
     2161 tagcgactaa tcgctagttc atttgctttg aaaacaacta attcagacat acatctcaat
     2221 tggtctaggt gattttaatc actataccaa ttgagatggg ctagtcaatg ataattacta
     2281 gtccttttcc tttgagttgt gggtatctgt aaattctgct agacctttgc tggaaaactt
     2341 gtaaattctg ctagaccctc tgtaaattcc gctagacctt tgtgtgtttt ttttgtttat
     2401 attcaagtgg ttataattta tagaataaag aaagaataaa aaaagataaa aagaatagat
     2461 cccagccctg tgtataactc actactttag tcagttccgc agtattacaa aaggatgtcg
     2521 caaacgctgt ttgctcctct acaaaacaga ccttaaaacc ctaaaggctt aagtagcacc
     2581 ctcgcaagct cgggcaaatc gctgaatatt ccttttgtct ccgaccatca ggcacctgag
     2641 tcgctgtctt tttcgtgaca ttcagttcgc tgcgctcacg gctctggcag tgaatggggg
     2701 taaatggcac tacaggcgcc ttttatggat tcatgcaagg aaactaccca taatacaaga
     2761 aaagcccgtc acgggcttct cagggcgttt tatggcgggt ctgctatgtg gtgctatctg
     2821 actttttgct gttcagcagt tcctgccctc tgattttcca gtctgaccac ttcggattat
     2881 cccgtgacag gtcattcaga ctggctaatg cacccagtaa ggcagcggta tcatcaacag
     2941 gcttacccgt cttactgtca accagacccg ccaggattct catgcataaa aaacgcccgg
     3001 cggcaaccga gcgttctgaa ttaattaatc atcgcgactg cagagatccc agagtcctta
     3061 ttagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat
     3121 accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg
     3181 ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa
     3241 tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac
     3301 gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc
     3361 gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt
     3421 acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag
     3481 cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg
     3541 agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc
     3601 agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg
     3661 cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac
     3721 cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg
     3781 tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc
     3841 atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcagatc ttgatcccct
     3901 gcgccatcag atccttggcg gcaagaaagc catccagttt actttgcagg gcttcccaac
     3961 cttaccag
//
