LOCUS       sequence-211118-        6808 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construct for expression of aequorin, calcium sensitive luciferase 
            reporter and hygromycin resistance gene by a bidirectional promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6808)
  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 6808)
  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-aequorin
FEATURES             Location/Qualifiers
     source          1..6808
                     /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(4522..5110)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4602..4621)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5281..6072,6073..6141))
                     /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     5904..5923
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6142..6246)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6283..6786
                     /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 ggcgcgtatg tcgggcggta
     2821 gcggaagtgg gggccagagc ggggggagtg gctccggagg acagtccggc gggagcgggt
     2881 caggcggtca atccggcggt agcgggtcgg gggggcaatc gggaggctcc ggttctgggg
     2941 gccagacgag caagtacgcc gtgaaattgg agcccgattt cgagaaccct cgatggatcg
     3001 ggcgtcataa gcacatgttt aacttcttgg acgtgaatca aaatggtaag attagcttag
     3061 acgagatggt ttacaaggca agtgacattg ttattaacaa tctaggcgca accccggagc
     3121 aagcgaagcg tcacaaggaa gcggtggagg cgttcttcgg cggagctggc atgaagtacg
     3181 gggtggagac cgagtggccc gagtacatca aggggtggaa gaagttggcc caaactgaac
     3241 tggacaagtt cgccaaaaac caggtcacac tcatacggct ctggggggac gcgctgcttg
     3301 atattattga caaggatcag aatggcgcta ttaccctgga cgagtggaag aagtacactc
     3361 aatcggctgg gattatccag agcgccgagg attgcgagga gacctttaag gtctgcgatc
     3421 ttgacgattc cgggcgtttg gatgcagacg aaatgacccg acagcatatc ggtttttggt
     3481 acacgatgga ccccgcctgt gagaagctct acggcggcgc ggtcccttga gctcgaaaga
     3541 tccaagagag acgagtagag attttttttt tgggattgat gtttgtcgtt ctttgagttg
     3601 tcgtcgagtt acgccttttg taagaatgtt ccgcaggaga ggaggaggat gggcatgagt
     3661 gagggtgaga gggcttgccc gctttttttt ttaaaaacgc tgaagacgtg gttgtcaaac
     3721 aaacccccca tagaaacgat tttgttacgg tgcggtccag acgtcacttg aatggctccg
     3781 cggaaaggcc agggagggaa ggggggaggg aggaaacatg aaacatgttg aacggctcaa
     3841 cagggtttgg gggacaagag aggtagcgcc ctgatggact gctccctccc ctcctttccc
     3901 tcaatgtctc attcatccat gcttccccct tctctctctc ccctccgttc catcccccgc
     3961 gggcgtggta gtggcgtgat gggatccact aaaatgtacg tgtaagaaaa gccggtgagc
     4021 ttacgctttt gtgaaagtgg gagtacgagt gttgtgtgtg tgtgtagtgg tttcagaccc
     4081 cagacagagg cgaagcagaa aaagcagacg atgaagacga cgaagaaatg agcagtctat
     4141 ttttatcgtg gaaacagaag aggtgatatc gtctcgttct ttgttatcac ctaccccgcg
     4201 tgcatgtaca tgcagccttt ttattttgta atctttcccg aaaaatcaac cgccacctcc
     4261 cccccgcctt ctctcaccca tcatcttctc ctgtttatct tctactttac actagatcgc
     4321 atggcacatc tccctcgcaa tccatcggtg caaccatcat cgatcccact cctccctccc
     4381 tccctccctc cctcctcgca tctcttctaa gaaatccgct agtgtttaaa ccacagaatc
     4441 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4501 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4561 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4621 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4681 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4741 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4801 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4861 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4921 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4981 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5041 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5101 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5161 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5221 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5281 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5341 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5401 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5461 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5521 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5581 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5641 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5701 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5761 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5821 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5881 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5941 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6001 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6061 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6121 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6181 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6241 tccgcgcaca tttccccgaa aagtgccacc tgggtccttt tcatcacgtg ctataaaaat
     6301 aattataatt taaatttttt aatataaata tataaattaa aaatagaaag taaaaaaaga
     6361 aattaaagaa aaaatagttt ttgttttccg aagatgtaaa agactctagg gggatcgcca
     6421 acaaatacta ccttttatct tgtacttcct gctctcaggt attaatgccg aattgtttca
     6481 tcttgtctgt gtagaagacc acacacgaaa atcctgtgat tttacatttt acttatcgtt
     6541 aatcgaatgt atatctattt aatctgcttt tcttgtctaa taaatatata tgtaaagtac
     6601 gctttttgtt gaaatttttt aaacctttgt ttattttttt ttcttcattc cgtaactctt
     6661 ctaccttctt tatttacttt ctaaaatcca aatacaaaac ataaaaataa ataaacacag
     6721 agtaaattcc caaattattc catcattaaa agatacgagg cgcgtgtaag ttacaggcaa
     6781 gcgatccgtc taagaaaccg gttaaggc
//
