LOCUS       sequence-245659-        6851 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construct for fusion expression of mNeonGreen reporter and G418 
            resistance gene by a bidirectional promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6851)
  AUTHORS   Poliner E, Cummings C, Newton L, Farre EM
  TITLE     Identification of circadian rhythms in Nannochloropsis species using
            bioluminescence reporter lines.
  JOURNAL   Plant J. 2019 Jul;99(1):112-127. doi: 10.1111/tpj.14314. Epub 2019 
            Apr 19.
  PUBMED    30883973
REFERENCE   2  (bases 1 to 6851)
  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: "10.1111/tpj.14314";
            journalName: "Plant J"; date: "2019-07"; volume: "99"; issue: "1"; 
            pages: "112-127"
COMMENT     Sequence Label: pNOC-ARS-stacked-NeoR-mNeonGreen
FEATURES             Location/Qualifiers
     source          1..6851
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(803..1597)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(803..1597)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(914..933)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1524..1543
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(1823..1834)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     rep_origin      complement(4565..5153)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4645..4664)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5324..6115,6116..6184))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5947..5966
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6185..6289)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6326..6829
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 taatggatgg gaagtcggta agtgaaaagg gctttgtgta aggtggatgt caaattttgc
       61 caacccgtgc ctctgtgttc gcagtgtagc aaggaagccc acaccaccgc ctcatattca
      121 aatcgaaatc atcgaagaaa tcggcgagga tctcatgccc caaagctgtc atcaaatgtg
      181 tcccaccaca cgaagatgcc gagtaaatga cgtgagttgt cctatgattg gcaggatgtg
      241 gcatttaaag tccaaaaggc cctgcttggt cggtcacctt ttgctcgcct ccccctcccg
      301 tcacacccta aacacgaatg cagaagaaaa tcatgtgggg aagcaacgat ttagaaatgg
      361 gaatggaggc tcttcacttc ctgaatctga ccaagctggc ggacacaacc gtagagggat
      421 ctttgacctg catcaataca ccgacatgtt aataaatcga gtaccaaaga aaagctactg
      481 aagtcagttt cggattgtgt tgtagaatct tttgagtcgt cactggaatg ctatgctcgc
      541 ttcaatcgtc gattagcttt gtaaattatt tctggtggca aggcggggtt caagtagcaa
      601 gtgggggcat ggcagcaccc ctccgagcct cgttcgttca ggtgtttttg ggacctttcc
      661 ctcccttcct gtcttcagac gcttttattt tcctttccgt tttctactag tgacaaaaaa
      721 agggcacaca ctctttactc gctcacaccc tcgtactcgg acacgcgctg ttaccgactc
      781 tggtacctca agcgcttcta gctcagaaga actcgtcaag aaggcgatag aaggcgatgc
      841 gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc
      901 caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac
      961 ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca
     1021 agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgctc gccttgagcc
     1081 tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga
     1141 caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga
     1201 atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata
     1261 ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata
     1321 gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg
     1381 tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca
     1441 ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat
     1501 cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg
     1561 ccggagaacc tgcgtgcaat ccatcttgtt caatcatggt cagctgctcg agggttgcgt
     1621 gtgtatctgt gtgcagtgga tattgttacc gagtttggtg agcgtgagtc cgttagtgcc
     1681 ctggtggtgg tggattagga gagtgggtga ctcggtgtcc atggctttct tcgctcatta
     1741 taggagggga aaggaatgag ggagggtggg gagaccgcgt ctgttgttga ccaccgattt
     1801 acttcttgcc tcccttcccc tccctcccct caatccgtac gacacaaata gtagccgagt
     1861 gtctgctgca gagcgcatga ttagtgtggt agacaacgag ggagggaagg atgtacaggg
     1921 catggcacgg agaagcgatg gtggccagga agaggagagg tcgtgagaac aggatgtgtt
     1981 gcgaatggat aaaaacagaa agcgatggct ctgggcttcg aaagcagggg acattaggac
     2041 gtgtagacca tctcgacgga tccctctgta tctctgttgt gcgtgaatgt tttctgtgca
     2101 cgtgtagtgt gtgagagtag aacccgggaa ctcgaacaga gaaaagcatg ggtggctgtg
     2161 gtgtggaggc ttcgttccca ccacatgccc ttctccttcg cctcgcctct ccctgccttc
     2221 ttccacgcac ccttgcgccc ctcgttctca atacctggct cacttccacc attcaaacaa
     2281 ccatcacgat acaggcattt atctatcgtt gaagacttct tcctccggta gatcttagcc
     2341 aaggtaagaa gaggggcatg cagcaaggag aaagaaatga tgcatgatga ggaatagaag
     2401 gggaggaggg agggatatga tgggaagcga aagcgcatat tctggtggtc tgctgcctga
     2461 tggggagtct agctgtgaca ctgaggacgg tggctgctgg tggctgcggg cgctgccttg
     2521 gtgatcaatg ggagtaaagg gagggaagga ggtagcgtga acggatgacg cggagaagtt
     2581 taggggtctc tttacgtatc gcccctgccg cccgcctctc tgcgataaat gtgcctgtta
     2641 ccctgcagcc tctattcttc actgtgttcc tgttttccaa cagcctctat tcttccctgt
     2701 cttttgttgc agtggcgtca tcctctcttt gccccagtcg tcgttctctc gactcactca
     2761 ctccccccct ccttccctcc ctccatccac agaatcgatt ggcgcgccct cgttaaccaa
     2821 ttggggagcg gcatggtctc taagggggaa gaagacaata tggcctcgct gcccgctacg
     2881 catgaactcc acatttttgg ttccatcaac ggtgtggatt tcgatatggt tggccaaggg
     2941 accggcaacc caaatgacgg ctatgaagag ttaaatctca agagcacgaa aggggatcta
     3001 cagtttagtc cgtggatact tgtgcctcac atcggatacg gatttcacca atacctgccc
     3061 taccctgacg gaatgagccc tttccaggcc gctatggttg atggaagtgg ttaccaggtc
     3121 catcggacga tgcaatttga ggatggggcg agcctgacgg tgaattacag atacacctat
     3181 gaggggagcc atataaaggg cgaagcccaa gtcaagggga ccgggtttcc agcggacgga
     3241 ccagtgatga caaatagttt gactgccgcg gattggtgtc gatccaaaaa gacatatcct
     3301 aacgacaaga cgataatcag tacgttcaaa tggtcctaca cgacgggtaa tggtaaacga
     3361 tataggagca cggcgcggac tacctatacc ttcgcaaaac cgatggctgc aaattacttg
     3421 aagaaccagc ctatgtatgt cttccgcaag accgagctga agcacagcaa gacggaactc
     3481 aattttaagg aatggcagaa agcctttacg gatgtgatgg ggatggatga gttgtataaa
     3541 ggaagcggca ggcctacgcg tgagtcgcga tgagctcgaa agatccaaga gagacgagta
     3601 gagatttttt ttttgggatt gatgtttgtc gttctttgag ttgtcgtcga gttacgcctt
     3661 ttgtaagaat gttccgcagg agaggaggag gatgggcatg agtgagggtg agagggcttg
     3721 cccgcttttt tttttaaaaa cgctgaagac gtggttgtca aacaaacccc ccatagaaac
     3781 gattttgtta cggtgcggtc cagacgtcac ttgaatggct ccgcggaaag gccagggagg
     3841 gaagggggga gggaggaaac atgaaacatg ttgaacggct caacagggtt tgggggacaa
     3901 gagaggtagc gccctgatgg actgctccct cccctccttt ccctcaatgt ctcattcatc
     3961 catgcttccc ccttctctct ctcccctccg ttccatcccc cgcgggcgtg gtagtggcgt
     4021 gatgggatcc actaaaatgt acgtgtaaga aaagccggtg agcttacgct tttgtgaaag
     4081 tgggagtacg agtgttgtgt gtgtgtgtag tggtttcaga ccccagacag aggcgaagca
     4141 gaaaaagcag acgatgaaga cgacgaagaa atgagcagtc tatttttatc gtggaaacag
     4201 aagaggtgat atcgtctcgt tctttgttat cacctacccc gcgtgcatgt acatgcagcc
     4261 tttttatttt gtaatctttc ccgaaaaatc aaccgccacc tcccccccgc cttctctcac
     4321 ccatcatctt ctcctgttta tcttctactt tacactagat cgcatggcac atctccctcg
     4381 caatccatcg gtgcaaccat catcgatccc actcctccct ccctccctcc ctccctcctc
     4441 gcatctcttc taagaaatcc gctagtgttt aaaccacaga atcaggggat aacgcaggaa
     4501 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4561 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4621 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4681 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4741 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4801 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4861 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4921 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4981 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5041 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5101 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5161 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5221 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5281 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5341 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5401 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5461 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5521 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5581 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5641 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5701 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5761 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5821 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5881 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5941 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6001 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6061 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6121 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6181 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6241 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6301 gaaaagtgcc acctgggtcc ttttcatcac gtgctataaa aataattata atttaaattt
     6361 tttaatataa atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag
     6421 tttttgtttt ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta
     6481 tcttgtactt cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag
     6541 accacacacg aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta
     6601 tttaatctgc ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt
     6661 tttaaacctt tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac
     6721 tttctaaaat ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta
     6781 ttccatcatt aaaagatacg aggcgcgtgt aagttacagg caagcgatcc gtctaagaaa
     6841 ccggttaagg c
//
