LOCUS       sequence-200533-        3965 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding mNeptune2.
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
FEATURES             Location/Qualifiers
     source          1..3965
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     38..56
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      1010..1056
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1229..2179)
                     /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      2227..2449
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(2955..2989)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(3030..3824)
                     /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(3141..3160)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3751..3770
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
       61 tcgtcttcac cacagctaac accacgtcgt ccctatctgc tgccctaggt ctatgagtgg
      121 ttgctggata actttacggg catgcataag gctcgtatga tatattcagg gagaccacaa
      181 cggtttccct ctacaaataa ttttgtttaa cttttctaga tttaagaagg agatatacat
      241 atgagtaaag gagaagaact gattaaggag aacatgcaca tgaagctgta catggagggc
      301 accgtgaaca accaccactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc
      361 acccagaccg gcagaatcaa ggtggtcgag ggcggccctt tacccttcgc cttcgacatc
      421 ctggctacct gcttcatgta cggcagcaag accttcatca accacaccca gggcatcccc
      481 gacttcttta agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa
      541 gacgggggcg tgctgaccgt tacccaggac accagcctgc aggacggctg cctgatctac
      601 aacgtcaagc tcagaggggt gaacttccca tccaacggcc ctgtgatgca gaagaaaaca
      661 ctgggctggg aggccagcac cgagaccctg taccccgctg acggcggcct ggaaggcaga
      721 tgcgacatgg ccctgaagtt agtgggcggg ggccacctgc actgcaactt gaagaccaca
      781 tacagatcca agaaacccgc taagaacttg aagatgcccg gcgtctactt cgtggacagg
      841 agactggaaa gaatcaagga ggccgacaat gagacctacg tcgagcagca cgaggtggct
      901 gtggccaggt actgcgacct gcctagcaaa ctggggcaca aacttaatgg catggatgaa
      961 ctatacaaat aaatgtccag acctgcaggc atgcaagctc tagaggcatc aaataaaacg
     1021 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgaaatgcac caaaaactcg
     1081 taaaagctct gatgtatcta tcttttttac accgttttca tctgtgcata tggacagttt
     1141 tccctttgat atctaacggt gaacagttgt tctacttttg tttgttagtc ttgatgcttc
     1201 actgatagat acaagagcca taagaacctc agatccttcc gtatttagcc agtatgttct
     1261 ctagtgtggt tcgttgtttt tgcgtgagcc atgagaacga accattgaga tcatgcttac
     1321 tttgcatgtc actcaaaaat tttgcctcaa aactggtgag ctgaattttt gcagttaaag
     1381 catcgtgtag tgtttttctt agtccgttac gtaggtagga atctgatgta atggttgttg
     1441 gtattttgtc accattcatt tttatctggt tgttctcaag ttcggttacg agatccattt
     1501 gtctatctag ttcaacttgg aaaatcaacg tatcagtcgg gcggcctcgc ttatcaacca
     1561 ccaatttcat attgctgtaa gtgtttaaat ctttacttat tggtttcaaa acccattggt
     1621 taagcctttt aaactcatgg tagttatttt caagcattaa catgaactta aattcatcaa
     1681 ggctaatctc tatatttgcc ttgtgagttt tcttttgtgt tagttctttt aataaccact
     1741 cataaatcct catagagtat ttgttttcaa aagacttaac atgttccaga ttatatttta
     1801 tgaatttttt taactggaaa agataaggca atatctcttc actaaaaact aattctaatt
     1861 tttcgcttga gaacttggca tagtttgtcc actggaaaat ctcaaagcct ttaaccaaag
     1921 gattcctgat ttccacagtt ctcgtcatca gctctctggt tgctttagct aatacaccat
     1981 aagcattttc cctactgatg ttcatcatct gagcgtattg gttataagtg aacgataccg
     2041 tccgttcttt ccttgtaggg ttttcaatcg tggggttgag tagtgccaca cagcataaaa
     2101 ttagcttggt ttcatgctcc gttaagtcat agcgactaat cgctagttca tttgctttga
     2161 aaacaactaa ttcagacata catctcaatt ggtctaggtg attttaatca ctataccaat
     2221 tgagatgggc tagtcaatga taattactag tccttttcct ttgagttgtg ggtatctgta
     2281 aattctgcta gacctttgct ggaaaacttg taaattctgc tagaccctct gtaaattccg
     2341 ctagaccttt gtgtgttttt tttgtttata ttcaagtggt tataatttat agaataaaga
     2401 aagaataaaa aaagataaaa agaatagatc ccagccctgt gtataactca ctactttagt
     2461 cagttccgca gtattacaaa aggatgtcgc aaacgctgtt tgctcctcta caaaacagac
     2521 cttaaaaccc taaaggctta agtagcaccc tcgcaagctc gggcaaatcg ctgaatattc
     2581 cttttgtctc cgaccatcag gcacctgagt cgctgtcttt ttcgtgacat tcagttcgct
     2641 gcgctcacgg ctctggcagt gaatgggggt aaatggcact acaggcgcct tttatggatt
     2701 catgcaagga aactacccat aatacaagaa aagcccgtca cgggcttctc agggcgtttt
     2761 atggcgggtc tgctatgtgg tgctatctga ctttttgctg ttcagcagtt cctgccctct
     2821 gattttccag tctgaccact tcggattatc ccgtgacagg tcattcagac tggctaatgc
     2881 acccagtaag gcagcggtat catcaacagg cttacccgtc ttactgtcaa ccagacccgc
     2941 caggattctc atgcataaaa aacgcccggc ggcaaccgag cgttctgaat taattaatca
     3001 tcgcgactgc agagatccca gagtccttat tagaagaact cgtcaagaag gcgatagaag
     3061 gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat
     3121 tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc
     3181 gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata
     3241 ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catgcgcgcc
     3301 ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc
     3361 tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg
     3421 tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg
     3481 atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg
     3541 cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga
     3601 acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cctgcagttc attcagggca
     3661 ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg
     3721 gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc
     3781 caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat
     3841 cctgtctctt gatcagatct tgatcccctg cgccatcaga tccttggcgg caagaaagcc
     3901 atccagttta ctttgcaggg cttcccaacc ttaccagagg gcgccccagc tggcaattcc
     3961 gacgt
//
