LOCUS       sequence-210253-        8790 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway compatible construct for N' terminal fusion or promoter 
            driven NanoLuciferase reporter, hygromycin resistance cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8790)
  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 8790)
  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-NLUC
FEATURES             Location/Qualifiers
     source          1..8790
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             801..1826
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             801..1826
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     protein_bind    2064..2188
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2213..2218,2219..2236,2237..2243)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2297..2956
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2363..2382)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3298..3603
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     3482..3501
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3644..3768)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             4304..4330
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(6292..6309)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6463..7051)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6543..6562)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7222..8013,8014..8082))
                     /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     7845..7864
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8083..8187)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8265..8720)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8402..8423)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8614..8633
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gtaggtctct aagatggagt ggatggagga ggaggcgagc gtagcagcaa gcgtgagtta
       61 tacagccagg cacatgtcgc aatccttcgg tctcgggctt aaaatccacg cactaatcac
      121 gctgggccat gcaaagagca atgccgaggc ccaccacaca aaacgctgtg tcgcgcgttg
      181 cggcctgaag cttcatactt cttagtcgcc gccaaaaggg ctcgagagac gagacccgtt
      241 ggcatgaccg atgttgttcg acgcggtttg cttcgtcaca gtcgacgtga ttcaggaatc
      301 tggagcctgc agatcatttt tttcagcctg atatcgttct tttccactga gaaccatcag
      361 accacctttt cttccattgt gtgaaggagt aggagttgcc gtgctgcttt gtgggagaca
      421 tctgcgatgg tgaccagcct cccgtcgtct ggtcgacgtg acgagcctct tcactgttct
      481 tcgacggaga gacgcaagcg agacggctct agaccttttg gacacgcatt ctgtgtgtga
      541 actagtggac agtgatacca cgtctgaaag ctcaccactg cccatggtgc agctacttgt
      601 cacaaagttt tgactccgtc ggtatcacca ttcgcgctcg tgtgcctggt tgttccgcca
      661 cgccggcctg ccccggggcg gggcaatatt ctaaaatctc acgcaaaaca ccgcacttac
      721 ccctcacaca tattcgtgat agaccaccac caatctcagc ccgcatcaac acagtcgagg
      781 gccccggggg gcaataagat atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt
      841 ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat
      901 ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg
      961 ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc gcgctcccga
     1021 ttccggaagt gcttgacatt ggggaattca gcgagagcct gacctattgc atctcccgcc
     1081 gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc
     1141 cggtcgcgga ggccatggat gcgatcgctg cggccgatct tagccagacg agcgggttcg
     1201 gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga
     1261 ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg
     1321 tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc
     1381 tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg
     1441 tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct
     1501 tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc
     1561 cggagcttgc aggatcgccg cggctccggg cgtatatgct ccgcattggt cttgaccaac
     1621 tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg
     1681 acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg
     1741 cggccgtctg gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca
     1801 gcactcgtcc gagggcaaag gaatagagta gatgccgacc ggagtccgca aaaatcacca
     1861 gtctctctct acaaatctat ctctctctat ttttctccag aataatgtgt gagtagttcc
     1921 cagataaggg aattagggtt cttatagggt ttcgctcatg tgttgagcat ataagaaacc
     1981 cttagtatgt atttgtattt gtaaaatact tctatcaata aaatttctaa ttcctaaaac
     2041 caaaatccag tgacccgata tcaacaagtt tgtacaaaaa agctgaacga gaaacgtaaa
     2101 atgatataaa tatcaatata ttaaattaga ttttgcataa aaaacagact acataatact
     2161 gtaaaacaca acatatccag tcactatggc ggccgcatta ggcaccccag gctttacact
     2221 ttatgcttcc ggctcgtata atgtgtggat tttgagttag gatccgtcga gattttcagg
     2281 agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca
     2341 atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca
     2401 gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt
     2461 ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg aattccgtat
     2521 ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt
     2581 ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca
     2641 gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc
     2701 taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag
     2761 ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa
     2821 atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt
     2881 ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg
     2941 gcagggcggg gcgtaaagat ctggatccgg cttactaaaa gccagataac agtatgcgta
     3001 tttgcgcgct gatttttgcg gtataagaat atatactgat atgtataccc gaagtatgtc
     3061 aaaaagaggt atgctatgaa gcagcgtatt acagtgacag ttgacagcga cagctatcag
     3121 ttgctcaagg catatatgat gtcaatatct ccggtctggt aagcacaacc atgcagaatg
     3181 aagcccgtcg tctgcgtgcc gaacgctgga aagcggaaaa tcaggaaggg atggctgagg
     3241 tcgcccggtt tattgaaatg aacggctctt ttgctgacga gaacaggggc tggtgaaatg
     3301 cagtttaagg tttacaccta taaaagagag agccgttatc gtctgtttgt ggatgtacag
     3361 agtgatatta ttgacacgcc cgggcgacgg atggtgatcc ccctggccag tgcacgtctg
     3421 ctgtcagata aagtctcccg tgaactttac ccggtggtgc atatcgggga tgaaagctgg
     3481 cgcatgatga ccaccgatat ggccagtgtg ccggtctccg ttatcgggga agaagtggct
     3541 gatctcagcc accgcgaaaa tgacatcaaa aacgccatta acctgatgtt ctggggaata
     3601 taaatgtcag gctcccttat acacagccag tctgcaggtc gaccatagtg actggatatg
     3661 ttgtgtttta cagtattatg tagtctgttt tttatgcaaa atctaattta atatattgat
     3721 atttatatca ttttacgttt ctcgttcagc tttcttgtac aaagtggttg atagcttggc
     3781 gcgcctcatg gtgtttactc tcgaggactt cgtgggcgac tggcggcaga cagctggcta
     3841 taacctcgac caggtgctgg agcagggcgg tgtgtcctcg cttttccaga acctgggcgt
     3901 ctccgtgacc ccaatccagc gcatcgtcct gagcggcgag aacggcctga agatcgacat
     3961 ccacgtcatc attccctacg agggcctgag cggggaccaa atgggccaga tcgagaagat
     4021 cttcaaggtt gtgtaccccg tggacgatca ccacttcaag gtgatactgc actacgggac
     4081 gctggtcatc gacggtgtga cgccgaacat gattgactac ttcggccgtc cttacgaggg
     4141 cattgccgtg ttcgacggca agaagatcac ggtgaccggc accctgtgga acggcaataa
     4201 gatcattgac gagcggctga tcaacccgga tggctcgctg ttgttccgcg tgaccatcaa
     4261 cggggtaacc ggctggcgcc tgtgcgagcg catcctggcc ggatacccct acgacgtgcc
     4321 cgactacgcg tgagctcgaa agatccaaga gagacgagta gagatttttt ttttgggatt
     4381 gatgtttgtc gttctttgag ttgtcgtcga gttacgcctt ttgtaagaat gttccgcagg
     4441 agaggaggag gatgggcatg agtgagggtg agagggcttg cccgcttttt tttttaaaaa
     4501 cgctgaagac gtggttgtca aacaaacccc ccatagaaac gattttgtta cggtgcggtc
     4561 cagacgtcac ttgaatggct ccgcggaaag gccagggagg gaagggggga gggaggaaac
     4621 atgaaacatg ttgaacggct caacagggtt tgggggacaa gagaggtagc gccctgatgg
     4681 actgctccct cccctccttt ccctcaatgt ctcattcatc catgcttccc ccttctctct
     4741 ctcccctccg ttccatcccc cgcgggcgtg gtagtggcgt gatgggatcc actaaaatgt
     4801 acgtgtaaga aaagccggtg agcttacgct tttgtgaaag tgggagtacg agtgttgtgt
     4861 gtgtgtgtag tggtttcaga ccccagacag aggcgaagca gaaaaagcag acgatgaaga
     4921 cgacgaagaa atgagcagtc tatttttatc gtggaaacag aagaggtgat atcgtctcgt
     4981 tctttgttat cacctacccc gcgtgcatgt acatgcagcc tttttatttt gtaatctttc
     5041 ccgaaaaatc aaccgccacc tcccccccgc cttctctcac ccatcatctt ctcctgttta
     5101 tcttctactt tacactagat cgcatggcac atctccctcg caatccatcg gtgcaaccat
     5161 catcgatccc actcctccct ccctccctcc ctccctcctc gcatctcttc taagaaatcc
     5221 gctagctgcg aacgcagctc acctacctca tttatcagag cctcgattcg gtccttttcc
     5281 ttctccccta cctcgccgca aacagcaaca gaagcaggag cagcaggagc agcggcacct
     5341 gtgtcaacag cacaagggca agtacagaca gaagacgagg cggaggagga ggaagacgat
     5401 gtgatgtcat tggagctggc taaactatta gccgttgctc tcaccgcctt tccccaaaac
     5461 tcttccaccc atcgtgtcac tccttctatc aggggaacca tccctctttt cctccccacc
     5521 ttcttcccct tgctcttctt cttcctttgt ctctttgttc cccctcctct cttcgcctcc
     5581 tcctccgcat cgtcctctcc ttttatcaag tcttgcgttg caaaggaagc aagctctttg
     5641 tgaaggaccg cgcaagggat aggagagtgg agagtgaggt gatgcagaca aaccgaggat
     5701 tcccgggcgt cagataacct aaaatcgact acttcttgag gtggaggagg agggggagga
     5761 gacggagaat aagtgtgtgt agaaatatag cttgctgcct ctgcttccat cttcgataaa
     5821 attgctgttg ctgttgctgt tgcggctgat atctgctgcg gctgctgctg cacctcctga
     5881 acgaagactg cagccgcgtc ttcaccctct tggactactc tgccctcctc ccttgccttt
     5941 acaactcgat cgacgagatt cgtgccagtg ctgtcatcat cattggaaag agcaacagca
     6001 tcaccttaag attgaatcct gttgccggtc ttgcgatgat tatcatataa tttctgttga
     6061 attacgttaa gcatgtaata attaacatgt aatgcatgac gttatttatg agatgggttt
     6121 ttatgattag agtcccgcaa ttatacattt aatacgcgat agaaaacaaa atatagcgcg
     6181 caaactagga taaattatcg cgcgcggtgt catctatgtt actaggctgc attaatgaat
     6241 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6301 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6361 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6421 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6481 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6541 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6601 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6661 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6721 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6781 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6841 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6901 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6961 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7021 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7081 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7141 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7201 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7261 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7321 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7381 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     7441 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     7501 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     7561 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     7621 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7681 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7741 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7801 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7861 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7921 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7981 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8041 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8101 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8161 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgatgcggt
     8221 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggaaattg taagcgttaa
     8281 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     8341 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     8401 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     8461 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     8521 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     8581 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     8641 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     8701 tgcgccgcta cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     8761 tcggtgcggg cctcttcgct attacgccag
//
