LOCUS       sequence-197022-        7379 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  deletes gnd locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7379)
  AUTHORS   Puszynska AM, O'Shea EK
  TITLE     Switching of metabolic programs in response to light availability is
            an essential function of the cyanobacterial circadian output 
            pathway.
  JOURNAL   Elife. 2017 Apr 21;6. pii: e23210. doi: 10.7554/eLife.23210.
  PUBMED    28430105
REFERENCE   2  (bases 1 to 7379)
  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"; journalName: "Elife."; 
            date: "2017-04-21"; volume: "6. pii: e23210. doi"; pages: " 
            10.7554/eLife.23210"
COMMENT     Sequence Label: gnd-
FEATURES             Location/Qualifiers
     source          1..7379
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     68..86
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(1166..1981)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1166..1981)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1889..1908
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1973..1992
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3207..4397
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(3559..3578)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             5036..5227
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(5221..5243)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5329..5469
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5384..5403
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5484..5501)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5655..6243)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5735..5754)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6414..7205,7206..7274))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7037..7056
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7275..7379)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
       61 ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa gaattctcat gtttgacagc
      121 ttatcatcga tagtctgtcg atcccagcag cgatagagat ggaggagttg atcgaaatag
      181 tgacgagcta attccagctg agggaaagac ttcaccgaaa gctgatcgct ccggcaatcc
      241 aaaacacgaa cttgatgatc ttcaatattg gcaactgcag gactgagaat gacgatccag
      301 cgaccgatat gttgtcgttg cgtttccata agaaagacag atgattgtgc gattaaagta
      361 gcgacaacaa ggctaacaaa tcgttaccta gggcacgatt ttgctctgaa ccagagtcag
      421 tcagcacaac gctaaacttt tgtagagctt tatcctcggc tcctaaacgg ttgcgatcgc
      481 catgttgatt cagcctgctg caacgacgtc taccccagag gtggctctcg caatcatcac
      541 ccaaggtcaa cagtggttat tgcagctgcg tgatgactta gattggattc tctatccggg
      601 gaagtggggc ttatttggcg gccatttaga accggatgaa actccccttg ccgccctcta
      661 tcgcgaactc caagaagaga ttggctacta cccctccgcc gcaaccccac tctggacgga
      721 aacagaagaa agggtcactc gctacctctt ccatgtgcct ttgacctgtg aactatcgac
      781 tttgcggctc ggcgaagggc aggatttctg cctcgtcgat gccgccgcga tcgcccaagg
      841 tcgagttttc tcgccccgtt tacagcagga acgagcgatc ggtgcgcccc accagcagat
      901 tttgctgcgc ttccaagggg agtattcgca caagcttgcc cccgtcatca gggcctgacc
      961 ggtgcgatcg ccgtcaagaa tccaccccca ggcattcaat ttttctctga tctgtaaata
     1021 aatccgcgat cgccgccgtg atccccgcaa agaaagtcta aagaggttgc cagagaaccc
     1081 caaattggca ggctaacctg aggtcattcg ttgtcctagc aggaattttc ctctgccagt
     1141 gttacaacca attaaccaat tctgattaga aaaactcatc gagcatcaaa tgaaactgca
     1201 atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag
     1261 gagaaaactc accgaggcag ttccatagga ttgcaagatc ctggtatcgg tctgcgattc
     1321 cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa
     1381 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagc ttatgcattt
     1441 ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa
     1501 ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa
     1561 aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa
     1621 caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga
     1681 tcgcagtggt tagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa
     1741 gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa
     1801 cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat
     1861 agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag
     1921 catccatgtt ggaatttaat cgcggcctcg agcaagacgt ttcccgttga atatggctca
     1981 taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat
     2041 ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtggctt tgttgaataa
     2101 atcgaacttt tgctgagttg aaggatcaga tcacgcatct tcccgacaac gcagggcgac
     2161 aaactccggt cgctgcgctg gtcagacgga ccaactcctt tggcagaatc aggactggca
     2221 aggcgatcgg agagcgtgat ggatgtgttg gtggtgcgcg gcattcgcgc ctacggctac
     2281 acggggttct ttgatgccga gcaagaactg ggccagtggt ttgaggtgga cttgcgcgtc
     2341 tgggtcgatc ttcagcctgc tggggacagc gatcagctcg gcgacaccct cgactacagc
     2401 caagcggtac aggtcgtgca aacggggatt cgccagagtc gctttcgcac gatcgagcga
     2461 ttggcggctg atctctgcga tcgcctgttt cagcagtttc cagccctacc ggaattggaa
     2521 ctgacgttgc ggaaactggc ccctcccatt cctgacttta gtggtgatgt cagcttggtt
     2581 ctgcggcgat cgcggaagtt aacgggtgcg gattgatgct agcaaggggt cgaagatgct
     2641 cgttctggag agctgacgag actgttttat agacttcaat caagagataa aagttgactc
     2701 gcgatcgtta gatttgggga tataatcctg agttctaata gctcttttta gggttccaaa
     2761 ctgtaagttt gaggattcac tttggaactt aagtcaatcc gaattgaatt cgggttggct
     2821 ctgattatcg tgatcaattg agacctagcc gcgatcgcaa atccatagaa cccgtctatt
     2881 cctatgacac tcgatttgct tgtagtcaac tctaaaaatt cgtgctagcg tctgatcgca
     2941 atcctcaatc ctcatgcagc ttcgcaagcg agatggattg ttgcttgtct tgaccgaagc
     3001 aaaggcttgt cgtagttcca gctgtcctca ttctctaggt cgttctgcta tgagcctcgc
     3061 tgcagtcttg gcgatcgcaa tttttgtggc ggtgattctg gctgcctcct ttgaactgac
     3121 cgatctcacg gtagtcgccc tggtcggagc gatgttaatg cggtagttta tcacagttaa
     3181 attgctaacg cagtcaggca ccgtgtatga aatctaacaa tgcgctcatc gtcatcctcg
     3241 gcaccgtcac cctggatgct gtaggcatag gcttggttat gccggtactg ccgggcctct
     3301 tgcgggatat cgtccattcc gacagcatcg ccagtcacta tggcgtgctg ctagcgctat
     3361 atgcgttgat gcaatttcta tgcgcacccg ttctcggagc actgtccgac cgctttggcc
     3421 gccgcccagt cctgctcgct tcgctacttg gagccactat cgactacgcg atcatggcga
     3481 ccacacccgt cctgtggatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca
     3541 caggtgcggt tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc
     3601 acttcgggct catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg
     3661 gactgttggg cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc
     3721 tcaacctact actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgaccga
     3781 tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg
     3841 tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc
     3901 tctgggtcat tttcggcgag gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc
     3961 ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt cgtcactggt cccgccacca
     4021 aacgtttcgg cgagaagcag gccattatcg ccggcatggc ggccgacgcg ctgggctacg
     4081 tcttgctggc gttcgcgacg cgaggctgga tggccttccc cattatgatt cttctcgctt
     4141 ccggcggcat cgggatgccc gcgttgcagg ccatgctgtc caggcaggta gatgacgacc
     4201 atcagggaca gcttcaagga tcgctcgcgg ctcttaccag cctaacttcg atcattggac
     4261 cgctgatcgt cacggcgatt tatgccgcct cggcgagcac atggaacggg ttggcatgga
     4321 ttgtaggcgc cgccctatac cttgtctgcc tccccgcgtt gcgtcgcggt gcatggagcc
     4381 gggccacctc gacctgaatg gaagccggcg gcacctcgct aacggattca ccactccaag
     4441 aattggagcc aatcaattct tgcggagaac tgtgaatgcg caaaccaacc cttggcagaa
     4501 catatccatc gcgtccgcca tctccagcag ccgcacgcgg cgcatctcgg gcagcgttgg
     4561 gtcctggcca cgggtgcgca tgatcgtgct cctgtcgttg aggacccggc taggctggcg
     4621 gggttgcctt actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg
     4681 ctgctgcaaa acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc
     4741 gtaaagtctg gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc
     4801 aggatgctgc tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg
     4861 accctgagtg atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca
     4921 acgttccagt aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg
     4981 tttcatcggt atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac
     5041 caaacaggaa aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct
     5101 tctggagaaa ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca
     5161 cgaccacgct gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa
     5221 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     5281 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac
     5341 ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt
     5401 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     5461 cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5521 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5581 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5641 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5701 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5761 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5821 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5881 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5941 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6001 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6061 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     6121 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6181 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6241 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6301 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6361 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6421 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6481 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6541 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6601 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6661 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6721 gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6781 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6841 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6901 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6961 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7021 ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7081 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7141 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7201 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7261 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7321 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcg
//
