LOCUS       sequence-201168-        3965 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding mNeptune2.5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3965)
  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 3965)
  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-mNeptune2.5
FEATURES             Location/Qualifiers
     source          1..3965
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      940..986
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1159..2109)
                     /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      2157..2379
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(2885..2919)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(2960..3754)
                     /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(3071..3090)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3681..3700
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3933..3951
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cacagctaac accacgtcgt ccctatctgc tgccctaggt ctatgagtgg ttgctggata
       61 actttacggg catgcataag gctcgtatga tatattcagg gagaccacaa cggtttccct
      121 ctacaaataa ttttgtttaa cttttctaga tttaagaagg agatatacat atgagtaaag
      181 gagaagaact gattaaggag aacatgcaca ccaagctgta catggagggc accgtgaaca
      241 accaccactt caagtgcaca cacgagggcg aaggcaagcc ctacgagggc acccagacca
      301 acagaatcaa ggtggtcgag ggcggccctt tacccttcgc cttcgacatc ctggctacct
      361 gcttcatgta cggcagcaag accttcatca accacaccca gggcatcccc gacttcttta
      421 agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa gacgggggcg
      481 tgctgaccgt tacccaggac accagcctgc aggacggctg cctgatctac aacgtcaagc
      541 tcagaggggt gaacttccca tccaacggcc ctgtgatgca gaagaaaaca ctgggctggg
      601 aggccagcac cgagaccctg taccccgctg acggcggcct ggaaggcaga tgcgacatgg
      661 ccctgaagtt agtgggcggg ggccacctgc actgcaactt gaagaccaca tacagatcca
      721 agaaacccgc taagaacttg aagatgcccg gcgtctactt cgtggacagg agactggaaa
      781 gaatcaagga ggccgacaat gagacctacg tcgagcagca cgaggtggct gtggccaggt
      841 actgcgacct gcctagcaaa ctggggcaca aacttaatgg catggatgaa ctatacaaat
      901 aaatgtccag acctgcaggc atgcaagctc tagaggcatc aaataaaacg aaaggctcag
      961 tcgaaagact gggcctttcg ttttatctgt tgaaatgcac caaaaactcg taaaagctct
     1021 gatgtatcta tcttttttac accgttttca tctgtgcata tggacagttt tccctttgat
     1081 atctaacggt gaacagttgt tctacttttg tttgttagtc ttgatgcttc actgatagat
     1141 acaagagcca taagaacctc agatccttcc gtatttagcc agtatgttct ctagtgtggt
     1201 tcgttgtttt tgcgtgagcc atgagaacga accattgaga tcatgcttac tttgcatgtc
     1261 actcaaaaat tttgcctcaa aactggtgag ctgaattttt gcagttaaag catcgtgtag
     1321 tgtttttctt agtccgttac gtaggtagga atctgatgta atggttgttg gtattttgtc
     1381 accattcatt tttatctggt tgttctcaag ttcggttacg agatccattt gtctatctag
     1441 ttcaacttgg aaaatcaacg tatcagtcgg gcggcctcgc ttatcaacca ccaatttcat
     1501 attgctgtaa gtgtttaaat ctttacttat tggtttcaaa acccattggt taagcctttt
     1561 aaactcatgg tagttatttt caagcattaa catgaactta aattcatcaa ggctaatctc
     1621 tatatttgcc ttgtgagttt tcttttgtgt tagttctttt aataaccact cataaatcct
     1681 catagagtat ttgttttcaa aagacttaac atgttccaga ttatatttta tgaatttttt
     1741 taactggaaa agataaggca atatctcttc actaaaaact aattctaatt tttcgcttga
     1801 gaacttggca tagtttgtcc actggaaaat ctcaaagcct ttaaccaaag gattcctgat
     1861 ttccacagtt ctcgtcatca gctctctggt tgctttagct aatacaccat aagcattttc
     1921 cctactgatg ttcatcatct gagcgtattg gttataagtg aacgataccg tccgttcttt
     1981 ccttgtaggg ttttcaatcg tggggttgag tagtgccaca cagcataaaa ttagcttggt
     2041 ttcatgctcc gttaagtcat agcgactaat cgctagttca tttgctttga aaacaactaa
     2101 ttcagacata catctcaatt ggtctaggtg attttaatca ctataccaat tgagatgggc
     2161 tagtcaatga taattactag tccttttcct ttgagttgtg ggtatctgta aattctgcta
     2221 gacctttgct ggaaaacttg taaattctgc tagaccctct gtaaattccg ctagaccttt
     2281 gtgtgttttt tttgtttata ttcaagtggt tataatttat agaataaaga aagaataaaa
     2341 aaagataaaa agaatagatc ccagccctgt gtataactca ctactttagt cagttccgca
     2401 gtattacaaa aggatgtcgc aaacgctgtt tgctcctcta caaaacagac cttaaaaccc
     2461 taaaggctta agtagcaccc tcgcaagctc gggcaaatcg ctgaatattc cttttgtctc
     2521 cgaccatcag gcacctgagt cgctgtcttt ttcgtgacat tcagttcgct gcgctcacgg
     2581 ctctggcagt gaatgggggt aaatggcact acaggcgcct tttatggatt catgcaagga
     2641 aactacccat aatacaagaa aagcccgtca cgggcttctc agggcgtttt atggcgggtc
     2701 tgctatgtgg tgctatctga ctttttgctg ttcagcagtt cctgccctct gattttccag
     2761 tctgaccact tcggattatc ccgtgacagg tcattcagac tggctaatgc acccagtaag
     2821 gcagcggtat catcaacagg cttacccgtc ttactgtcaa ccagacccgc caggattctc
     2881 atgcataaaa aacgcccggc ggcaaccgag cgttctgaat taattaatca tcgcgactgc
     2941 agagatccca gagtccttat tagaagaact cgtcaagaag gcgatagaag gcgatgcgct
     3001 gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa
     3061 gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca
     3121 gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc
     3181 aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg
     3241 cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa
     3301 gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg
     3361 ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt
     3421 tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca
     3481 gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg
     3541 tggccagcca cgatagccgc gctgcctcgt cctgcagttc attcagggca ccggacaggt
     3601 cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag
     3661 agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg
     3721 gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt
     3781 gatcagatct tgatcccctg cgccatcaga tccttggcgg caagaaagcc atccagttta
     3841 ctttgcaggg cttcccaacc ttaccagagg gcgccccagc tggcaattcc gacgtctaag
     3901 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     3961 ttcac
//
