LOCUS       sequence-197033-        7887 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  drives expression of rpaA from an rpaA-independent promoter 
            (kanamycin resistance casette).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7887)
  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 7887)
  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: P0050::rpaA (KmR)
FEATURES             Location/Qualifiers
     source          1..7887
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     68..86
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             227..238
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     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
                     SYFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     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             3715..4905
                     /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(4067..4086)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             5544..5735
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(5729..5751)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5837..5977
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5892..5911
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5992..6009)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6163..6751)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6243..6262)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6922..7713,7714..7782))
                     /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     7545..7564
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7783..7887)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
       61 ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa gaattctcat gtttgacagc
      121 ttatcatcga tacctcctag aagctcatca gcgagatctc tggcaacagg tcagtccggc
      181 actccttgat caactgcgat cgctttccct tgaagcagaa gctgatattg aaggtcgact
      241 aggaagtatt ccagataaca agttgcgcta gttggacttt attacaagca gttgcaatct
      301 tcggacgatt ctgctgtcgc tggttaccgt agaactattc ctaatgcttg ctcaatccct
      361 atgagttcac ttgttgttgt cggatttccc aattcgcaag aagccgaaga agtccgaatc
      421 aaacttgctc agctacagac agaacattta attgccctag aagatgctgt tgttgtcagc
      481 aaggattccg agggccatgt caaactcaat caagcgatta atttaacggc aactggtgcc
      541 gtcagcggcg gtttttgggg cacccttgtc ggcttgatct tcctcaatcc cttggcgggt
      601 gcggcgatcg gtgcagcggc tggtgcactc agcggttcct tgagcgacat tggcatcaac
      661 gatgactttc tgcgcgaggt cggcgaaacc ctgcaggtcg gatcagccgc cctctgcctc
      721 ctcatccgcg aagcaacccc cgatcgcatc ctgaaagagt tgcgagtcta cgccaaagac
      781 gcccggattc tacaaacgaa cttgagcttt gccgatgaag aaaacctcaa ggcagcccta
      841 gaaaaagcca aagccgaagc ccaagccctg cgtctcaatt aatctccaac gcttcgatcg
      901 ctcctgatta cttgagttga tcaaggctca gatcaagagc gatcgctact tcaatacagc
      961 ttacttttgg ccgctatctc aacagcagtt caatcctaag aaactgcaga agcagggatt
     1021 gagtaatttg atggttgact gcagtcctta tcagttgctt ctcaacaatt atccaaccgc
     1081 aattacggac agagcactgg actagcacca ctgctaaaac cgcaaagctc ggctgccagt
     1141 gttacaacca attaaccaat tctgattaga aaaactcatc gagcatcaaa tgaaactgca
     1201 atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag
     1261 gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc
     1321 cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa
     1381 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagc ttatgcattt
     1441 ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaataa ctcgcatcaa
     1501 ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa
     1561 aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa
     1621 caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga
     1681 tcgcagtggt gagtaaccat 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 gcaggctgcc
     2161 tttggttgca ataacggtaa atttaccgtc tgtcccgatt gtgccagtca cgttaatatt
     2221 cgctgcttta ccacttccag cagccgttac ttggctagca ttgccgcaag ttatgttaaa
     2281 gttacctgca ctgctaccaa taccaagctg tgtacaagac gcattggtga cgtagttagg
     2341 atctagtgtc gccgcactaa gggtacgaag agcagcaccg actgctgatt tgccttcaga
     2401 ggcataggct ttatctgttt gatccaacag gttcggtaat gcaactgctg ccaagatgcc
     2461 aacgataatg ataacgacca gcaactcgac aagggtaaaa cctttttctt cttcaccttg
     2521 ttcgtcttta cgctgggcaa gatattgtag ataacgaaga cggagggagt cagacattca
     2581 ggagatcctc agcaagagag ctgttcttga agtaacttac ccctacagca ggcttttatt
     2641 tcaggttcct gaaaatttcc gcatgaaacc ccgcatcctc gtgatcgatg atgactcagc
     2701 catcttggag ctggtcgccg tcaatctgga gatgtctggc tatgacgtac gcaaagctga
     2761 ggacggcatt aaaggtcagg ctttagctgt tcagctagtt cccgacctga tcatgctgga
     2821 tctaatgctg ccgcgggttg atggctttac cgtctgtcag cgactgcggc gcgatgagcg
     2881 tactgccgaa attccggtgc tgatgctgac cgccctcgga cagactcagg ataaggttga
     2941 aggcttcaac gcgggtgctg acgattatct gactaagccc ttcgaagttg aagagatgct
     3001 ggcccgcgtg cgtgccttgc tgcggcgcac cgatcgcatt ccccatgcag cccgccatag
     3061 cgaaattctc agctacggtc cgctgaccct gattcccgag cggtttgagg ccatttggtt
     3121 caaccgcacg gtcaagctga ctcacttgga atttgagttg ttgcactgcc tgttgcaacg
     3181 ccacggccaa acggttgcgc cgagcgaaat cctcaaagaa gtctggggct atgatcccga
     3241 cgatgacatc gagacgattc gcgtccacat ccgtcatctg cgcaccaagc tcgagcccga
     3301 tccccggcac ccgcgctaca tcaaaacggt ctatggagcg ggctactgcc ttgagctgcc
     3361 ggccgagacg gaactccacc aacacgccga tcagtttcct tcggcgtcct gaacccaagc
     3421 actgagcagc acttcccaga cgagtcgcgg ctgaacaaag gctcgcagtt gtttctgtgc
     3481 ctgttcgagc tggagcagtt gctgggactg cagctgttgc tgggaccagc cctgctgctg
     3541 gaggtagcga caaagccaaa gctgggcttc gacgtcgagg ccagcatcga tcgcttgcgc
     3601 cagttccaaa gccacttgca aagagtttgg ccgttgctga aggcgatcgc gcaattctgg
     3661 cgttaatgcg gtagtttatc acagttaaat tgctaacgca gtcaggcacc gtgtatgaaa
     3721 tctaacaatg cgctcatcgt catcctcggc accgtcaccc tggatgctgt aggcataggc
     3781 ttggttatgc cggtactgcc gggcctcttg cgggatatcg tccattccga cagcatcgcc
     3841 agtcactatg gcgtgctgct agcgctatat gcgttgatgc aatttctatg cgcacccgtt
     3901 ctcggagcac tgtccgaccg ctttggccgc cgcccagtcc tgctcgcttc gctacttgga
     3961 gccactatcg actacgcgat catggcgacc acacccgtcc tgtggatcct ctacgccgga
     4021 cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg ctggcgccta tatcgccgac
     4081 atcaccgatg gggaagatcg ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg
     4141 ggtatggtgg caggccccgt ggccggggga ctgttgggcg ccatctcctt gcatgcacca
     4201 ttccttgcgg cggcggtgct caacggcctc aacctactac tgggctgctt cctaatgcag
     4261 gagtcgcata agggagagcg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc
     4321 ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg
     4381 caactcgtag gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc
     4441 tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca cgccctcgct
     4501 caagccttcg tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc
     4561 ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg
     4621 gccttcccca ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc
     4681 atgctgtcca ggcaggtaga tgacgaccat cagggacagc ttcaaggatc gctcgcggct
     4741 cttaccagcc taacttcgat cattggaccg ctgatcgtca cggcgattta tgccgcctcg
     4801 gcgagcacat ggaacgggtt ggcatggatt gtaggcgccg ccctatacct tgtctgcctc
     4861 cccgcgttgc gtcgcggtgc atggagccgg gccacctcga cctgaatgga agccggcggc
     4921 acctcgctaa cggattcacc actccaagaa ttggagccaa tcaattcttg cggagaactg
     4981 tgaatgcgca aaccaaccct tggcagaaca tatccatcgc gtccgccatc tccagcagcc
     5041 gcacgcggcg catctcgggc agcgttgggt cctggccacg ggtgcgcatg atcgtgctcc
     5101 tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac
     5161 cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa
     5221 catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct
     5281 gcaccattat gttccggatc tgcatcgcag gatgctgctg gctaccctgt ggaacaccta
     5341 catctgtatt aacgaagcgc tggcattgac cctgagtgat ttttctctgg tcccgccgca
     5401 tccataccgc cagttgttta ccctcacaac gttccagtaa ccgggcatgt tcatcatcag
     5461 taacccgtat cgtgagcatc ctctctcgtt tcatcggtat cattaccccc atgaacagaa
     5521 atccccctta cacggaggca tcagtgacca aacaggaaaa aaccgccctt aacatggccc
     5581 gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg
     5641 aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc
     5701 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     5761 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     5821 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg
     5881 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     5941 accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac
     6001 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6061 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6121 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6181 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6241 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6301 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6361 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6421 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6481 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6541 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6601 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6661 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6721 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6781 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6841 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6901 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6961 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7021 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7081 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     7141 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     7201 gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc
     7261 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     7321 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7381 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7441 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7501 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca
     7561 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7621 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7681 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7741 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7801 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7861 gaaaaataaa caaatagggg ttccgcg
//
