LOCUS       sequence-245653-        7355 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construct for fusion expression of GeNL reporter and neomycin/G418 
            resistance gene by a bidirectional promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7355)
  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 7355)
  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-stacked-GeNL
FEATURES             Location/Qualifiers
     source          1..7355
                     /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"
     CDS             4018..4044
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      complement(5069..5657)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5149..5168)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5828..6619,6620..6688))
                     /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     6451..6470
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6689..6793)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6830..7333
                     /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 gggttcgagg acttcgtggg cgactggcgg
     3541 cagacggctg gctataacct cgaccaggtg ctggagcagg gcggtgtgtc ctcgcttttc
     3601 cagaacctgg gcgtctccgt gaccccaatc cagcgcatcg tcctgagcgg cgagaacggc
     3661 ctgaagatcg acatccacgt catcattccc tacgagggcc tgagcgggga ccaaatgggc
     3721 cagatcgaga agatcttcaa ggttgtgtac cccgtggacg atcaccactt caaggtgata
     3781 ctgcactacg ggacgctggt catcgacggt gtgacgccga acatgattga ctacttcggc
     3841 cgtccttacg agggcattgc cgtgttcgac ggcaagaaga tcacggtgac cggcaccctg
     3901 tggaacggca ataagatcat tgacgagcgg ctgatcaacc cggatggctc gctgttgttc
     3961 cgcgtgacca tcaacggggt aaccggctgg cgcctgtgcg agcgcatcct ggccggatac
     4021 ccctacgacg tgcccgacta cgcgggaagc ggcaggccta cgcgtgagtc gcgatgagct
     4081 cgaaagatcc aagagagacg agtagagatt ttttttttgg gattgatgtt tgtcgttctt
     4141 tgagttgtcg tcgagttacg ccttttgtaa gaatgttccg caggagagga ggaggatggg
     4201 catgagtgag ggtgagaggg cttgcccgct ttttttttta aaaacgctga agacgtggtt
     4261 gtcaaacaaa ccccccatag aaacgatttt gttacggtgc ggtccagacg tcacttgaat
     4321 ggctccgcgg aaaggccagg gagggaaggg gggagggagg aaacatgaaa catgttgaac
     4381 ggctcaacag ggtttggggg acaagagagg tagcgccctg atggactgct ccctcccctc
     4441 ctttccctca atgtctcatt catccatgct tcccccttct ctctctcccc tccgttccat
     4501 cccccgcggg cgtggtagtg gcgtgatggg atccactaaa atgtacgtgt aagaaaagcc
     4561 ggtgagctta cgcttttgtg aaagtgggag tacgagtgtt gtgtgtgtgt gtagtggttt
     4621 cagaccccag acagaggcga agcagaaaaa gcagacgatg aagacgacga agaaatgagc
     4681 agtctatttt tatcgtggaa acagaagagg tgatatcgtc tcgttctttg ttatcaccta
     4741 ccccgcgtgc atgtacatgc agccttttta ttttgtaatc tttcccgaaa aatcaaccgc
     4801 cacctccccc ccgccttctc tcacccatca tcttctcctg tttatcttct actttacact
     4861 agatcgcatg gcacatctcc ctcgcaatcc atcggtgcaa ccatcatcga tcccactcct
     4921 ccctccctcc ctccctccct cctcgcatct cttctaagaa atccgctagt gtttaaacca
     4981 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5041 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5101 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5161 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5221 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5281 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5341 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5401 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5461 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5521 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5581 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5641 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5701 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5761 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5821 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5881 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5941 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6001 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6061 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6121 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6181 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6241 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6301 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6361 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6421 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6481 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6541 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6601 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6661 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6721 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6781 taggggttcc gcgcacattt ccccgaaaag tgccacctgg gtccttttca tcacgtgcta
     6841 taaaaataat tataatttaa attttttaat ataaatatat aaattaaaaa tagaaagtaa
     6901 aaaaagaaat taaagaaaaa atagtttttg ttttccgaag atgtaaaaga ctctaggggg
     6961 atcgccaaca aatactacct tttatcttgt acttcctgct ctcaggtatt aatgccgaat
     7021 tgtttcatct tgtctgtgta gaagaccaca cacgaaaatc ctgtgatttt acattttact
     7081 tatcgttaat cgaatgtata tctatttaat ctgcttttct tgtctaataa atatatatgt
     7141 aaagtacgct ttttgttgaa attttttaaa cctttgttta tttttttttc ttcattccgt
     7201 aactcttcta ccttctttat ttactttcta aaatccaaat acaaaacata aaaataaata
     7261 aacacagagt aaattcccaa attattccat cattaaaaga tacgaggcgc gtgtaagtta
     7321 caggcaagcg atccgtctaa gaaaccggtt aaggc
//
