LOCUS       sequence-196985-        7477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  drives expression of rpaA D53A variant from an rpaA-independent 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7477)
  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 7477)
  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::rpaAD53A
FEATURES             Location/Qualifiers
     source          1..7477
                     /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"
     promoter        join(1171..1176,1177..1193,1194..1199)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             1388..1921
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     CDS             3305..4495
                     /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(3657..3676)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             5134..5325
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(5319..5341)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5427..5567
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5482..5501
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5582..5599)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5753..6341)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5833..5852)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6512..7303,7304..7372))
                     /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     7135..7154
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7373..7477)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
       61 ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa gaattctcat gtttgacagc
      121 ttatcatcga tacctcctag aacctcatca 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 gggacgcaca
     1141 ccgtggaaac ggatgaaggc acgaacccag ttgacataag cctgttcggt tcgtaaactg
     1201 taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt
     1261 ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgt acagtctatg
     1321 cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc
     1381 agcaacgatg ttacgcagca gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa
     1441 gttaggtggc tcaagtatgg gcatcattcg cacatgtagg ctcggccctg accaagtcaa
     1501 atccatgcgg gctgctcttg atcttttcgg tcgtgagttc ggagacgtag ccacctactc
     1561 ccaacatcag ccggactccg attacctcgg gaacttgctc cgtagtaaga cattcatcgc
     1621 gcttgctgcc ttcgaccaag aagcggttgt tggcgctctc gcggcttacg ttctgcccaa
     1681 gtttgagcag ccgcgtagtg agatctatat ctatgatctc gcagtctccg gcgagcaccg
     1741 gaggcagggc attgccaccg cgctcatcaa tctcctcaag catgaggcca acgcgcttgg
     1801 tgcttatgtg atctacgtgc aagcagatta cggtgacgat cccgcagtgg ctctctatac
     1861 aaagttgggc atacgggaag aagtgatgca ctttgatatc gacccaagta ccgccaccta
     1921 acaattcgtt caagccgaga tcggcttccc ggccgcggag ttgttcggta aattgtcaca
     1981 acgccgcact agtggctgcc tttggttgca ataacggtaa atttaccgtc tgtcccgatt
     2041 gtgccagtca cgttaatatt cgctgcttta ccacttccag cagccgttac ttggctagca
     2101 ttgccgcaag ttatgttaaa gttacctgca ctgctaccaa taccaagctg tgtacaagac
     2161 gcattggtga cgtagttagg atctagtgtc gccgcactaa gggtacgaag agcagcaccg
     2221 actgctgatt tgccttcaga ggcataggct ttatctgttt gatccaacag gttcggtaat
     2281 gcaactgctg ccaagatgcc aacgataatg ataacgacca gcaactcgac aagggtaaaa
     2341 cctttttctt cttcaccttg ttcgtcttta cgctgggcaa gatattgtag ataacgaaga
     2401 cggagggagt cagacattca ggagatcctc agcaagagag ctgttcttga agtaacttac
     2461 ccctacagca ggcttttatt tcaggttcct gaaaatttcc gcatgaaacc ccgcatcctc
     2521 gtgatcgatg atgactcagc catcttggag ctggtcgccg tcaatctgga gatgtctggc
     2581 tatgacgtac gcaaagctga ggacggcatt aaaggtcagg ctttagctgt tcagctagtt
     2641 cccgacctga tcatgctggc tctaatgctg ccgcgggttg atggctttac cgtctgtcag
     2701 cgactgcggc gcgatgagcg tactgccgaa attccggtgc tgatgctgac cgccctcgga
     2761 cagactcagg ataaggttga aggcttcaac gcgggtgctg acgattatct gactaagccc
     2821 ttcgaagttg aagagatgct ggcccgcgtg cgtgccttgc tgcggcgcac cgatcgcatt
     2881 ccccatgcag cccgccatag cgaaattctc agctacggtc cgctgaccct gattcccgag
     2941 cggtttgagg ccatttggtt caaccgcacg gtcaagctga ctcacttgga atttgagttg
     3001 ttgcactgcc tgttgcaacg ccacggccaa acggttgcgc cgagcgaaat cctcaaagaa
     3061 gtctggggct atgatcccga cgatgacatc gagacgattc gcgtccacat ccgtcatctg
     3121 cgcaccaagc tcgagcccga tccccggcac ccgcgctaca tcaaaacggt ctatggagcg
     3181 ggctactgcc ttgagctgcc ggccgagacg gaactccacc aacacgccga tcagtttcct
     3241 tcggcgtcct gattaatgcg gtagtttatc acagttaaat tgctaacgca gtcaggcacc
     3301 gtgtatgaaa tctaacaatg cgctcatcgt catcctcggc accgtcaccc tggatgctgt
     3361 aggcataggc ttggttatgc cggtactgcc gggcctcttg cgggatatcg tccattccga
     3421 cagcatcgcc agtcactatg gcgtgctgct agcgctatat gcgttgatgc aatttctatg
     3481 cgcacccgtt ctcggagcac tgtccgaccg ctttggccgc cgcccagtcc tgctcgcttc
     3541 gctacttgga gccactatcg actacgcgat catggcgacc acacccgtcc tgtggatcct
     3601 ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg ctggcgccta
     3661 tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca tgagcgcttg
     3721 tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg ccatctcctt
     3781 gcatgcacca ttccttgcgg cggcggtgct caacggcctc aacctactac tgggctgctt
     3841 cctaatgcag gagtcgcata agggagagcg tcgaccgatg cccttgagag ccttcaaccc
     3901 agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt
     3961 ctttatcatg caactcgtag gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga
     4021 ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca
     4081 cgccctcgct caagccttcg tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc
     4141 cattatcgcc ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt tcgcgacgcg
     4201 aggctggatg gccttcccca ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc
     4261 gttgcaggcc atgctgtcca ggcaggtaga tgacgaccat cagggacagc ttcaaggatc
     4321 gctcgcggct cttaccagcc taacttcgat cattggaccg ctgatcgtca cggcgattta
     4381 tgccgcctcg gcgagcacat ggaacgggtt ggcatggatt gtaggcgccg ccctatacct
     4441 tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga cctgaatgga
     4501 agccggcggc acctcgctaa cggattcacc actccaagaa ttggagccaa tcaattcttg
     4561 cggagaactg tgaatgcgca aaccaaccct tggcagaaca tatccatcgc gtccgccatc
     4621 tccagcagcc gcacgcggcg catctcgggc agcgttgggt cctggccacg ggtgcgcatg
     4681 atcgtgctcc tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag
     4741 aatgaatcac cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc
     4801 tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag
     4861 tcagcgccct gcaccattat gttccggatc tgcatcgcag gatgctgctg gctaccctgt
     4921 ggaacaccta catctgtatt aacgaagcgc tggcattgac cctgagtgat ttttctctgg
     4981 tcccgccgca tccataccgc cagttgttta ccctcacaac gttccagtaa ccgggcatgt
     5041 tcatcatcag taacccgtat cgtgagcatc ctctctcgtt tcatcggtat cattaccccc
     5101 atgaacagaa atccccctta cacggaggca tcagtgacca aacaggaaaa aaccgccctt
     5161 aacatggccc gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg
     5221 gacgcggatg aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac
     5281 cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     5341 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     5401 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
     5461 gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     5521 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt
     5581 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5641 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5701 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5761 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5821 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5881 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5941 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6001 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6061 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6121 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6181 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6241 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6301 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6361 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6421 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6481 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6541 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6601 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6661 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6721 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6781 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg
     6841 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6901 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6961 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7021 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7081 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata
     7141 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7201 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7261 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7321 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7381 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7441 aatgtattta gaaaaataaa caaatagggg ttccgcg
//
