LOCUS       sequence-197032-        6649 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  deletes rpaA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6649)
  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 6649)
  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: rpaA-
FEATURES             Location/Qualifiers
     source          1..6649
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        join(505..510,511..527,528..533)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             722..1255
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     complement(2762..2778)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2987..3006)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3106..3128
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3166..3184)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3252..3356
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3357..3425,3426..4217)
                     /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     complement(3575..3594)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4388..4976
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4877..4896
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5130..5147
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5264..5285
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5300..5305,5306..5323,5324..5330)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5338..5354
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5343..5365
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5362..5378
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5362..5378
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5470..5481
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 gtaaaacgac ggccagtgcc aagctttcgc gagctcgaga tctagatctg ctcgcgcagg
       61 ctgggtgcca agctctcggg taacatcaag gcccgatcct tggagccctt gccctcccgc
      121 acgatgatcg tgccgtgatc gaaatccaga tccttgaccc gcagttgcaa accctcactg
      181 atccgcatgc ccgttccata cagaagctgg gcgaacaaac gatgctcgcc ttccagaaaa
      241 ccgaggatgc gaaccacttc atccggggtc agcaccaccg gcaagcgccg cgacggccga
      301 ggtcttccga tctcctgaag ccagggcaga tccgtgcaca gcaccttgcc gtagaagaac
      361 agcaaggccg ccaatgcctg acgatgcgtg gagaccgaaa ccttgcgctc gttcgccagc
      421 caggacagaa atgcctcgac ttcgctgctg cccaaggttg ccgggtgacg cacaccgtgg
      481 aaacggatga aggcacgaac ccagttgaca taagcctgtt cggttcgtaa actgtaatgc
      541 aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac
      601 ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttgtacagtc tatgcctcgg
      661 gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac
      721 gatgttacgc agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttagg
      781 tggctcaagt atgggcatca ttcgcacatg taggctcggc cctgaccaag tcaaatccat
      841 gcgggctgct cttgatcttt tcggtcgtga gttcggagac gtagccacct actcccaaca
      901 tcagccggac tccgattacc tcgggaactt gctccgtagt aagacattca tcgcgcttgc
      961 tgccttcgac caagaagcgg ttgttggcgc tctcgcggct tacgttctgc ccaagtttga
     1021 gcagccgcgt agtgagatct atatctatga tctcgcagtc tccggcgagc accggaggca
     1081 gggcattgcc accgcgctca tcaatctcct caagcatgag gccaacgcgc ttggtgctta
     1141 tgtgatctac gtgcaagcag attacggtga cgatcccgca gtggctctct atacaaagtt
     1201 gggcatacgg gaagaagtga tgcactttga tatcgaccca agtaccgcca cctaacaatt
     1261 cgttcaagcc gagatcggct tcccggccgc ggagttgttc ggtaaattgt cacaacgccg
     1321 cggccgcaaa gcgctccggc ttaactcagg cgttagtccc atcacccgct ttatgcgcgc
     1381 agccacttat ttcagcgctc cgccctgttc caaaggcttt ctagcctttt gggcctccag
     1441 cgcttgtgca gcaagcgcca ccagctatcc ttcttgcagc gctgcgccgc tttcgtggtc
     1501 cagcggtttt tcttgggctg cgcccttctt caaggccggg cgctcccttt tggccttcgg
     1561 gtctaactct gcggtcaagc ggacccgcat tctgcgggcc gcttaccttg gccgttagac
     1621 atcatgaggg tagcggtgac tcgatgaatt cccatcggaa ctccaccaac acgccgatca
     1681 gtttccttcg gcgtcctgaa cccaagcact gagcagcact tcccagacga gtcgcggctg
     1741 aacaaaggct cgcagttgtt tctgtgcctg ttcgagctgg agcagttgct gggactgcag
     1801 ctgttgctgg gaccagccct gctgctggag gtagcgacaa agccaaagct gggcttcgac
     1861 gtcgaggcca gcatcgatcg cttgcgccag ttccaaagcc acttgcaaag agtttggccg
     1921 ttgctgaagg cgatcgcgca attctggcgg gattgcttgc agttgttgcc aatggctgat
     1981 agctgctcct ggactgcctt gagctaaggc cagaaccgct ggcgcgtcga gaatttctgt
     2041 aaagccgtgg gtctgcaaaa tttgcgccat cgcctgctca gaaaggcgat agaagggaat
     2101 tcgctggcaa cgagaaacaa gggttggcag tagcagctca ggactcggtg ccatcaggat
     2161 aatcgtggct tgaccgggct cttctagcgt tttgagcaac ccattggctg ccgcctcagc
     2221 catcgtttca gcctgttcaa tcaccaccag tgatcgcggg gcagcgagag gaggtcgtgc
     2281 cagaaactga gtgatttggc gaatttgctc cagtcgcact tggggcggtg ttcgccgctg
     2341 aattccctgg gcttgggctt gacttgcagg aatcaaggtt ccctgctggg aataagtcgg
     2401 ctcaacccag agcagatcgg gatgattgcc gcgagcgacc cgtcccgact gatcagccaa
     2461 gaggagcggt gcccaacaac gggcagccag tccacgaccg gtgccatcag gaccagcaaa
     2521 caggtaagcc ggcgccaagc gatcgcgatc gcaggctgcc agcagcaact gaacggcaag
     2581 gtcttgaccg atgagcggcg caaagggatt catcaccggg gccagcgctg ttgcaccacg
     2641 gcttgaatac gagcggcaac ctgatcagga tctccagctg catcgatgcg aaccacccga
     2701 tcaggctctg ccgccgcgat cgcctgaaat ccttgacgca gggatccacg tcgagcttgg
     2761 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     2821 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     2881 gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc
     2941 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg
     3001 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3061 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3121 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     3181 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     3241 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3301 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     3361 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3421 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     3481 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3541 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3601 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3661 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3721 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3781 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3841 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3901 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     3961 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     4021 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     4081 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4141 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4201 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4261 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4321 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4381 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4441 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4501 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4561 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4621 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4681 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4741 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4801 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4861 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4921 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4981 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5041 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     5101 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     5161 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     5221 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     5281 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     5341 tgagcggata acaatttcac acaggaaaca gctatgacat gattacgaat tcgacgtgga
     5401 tcctggcaac aggtcagtcc ggcactcctt gatcaactgc gatcgctttc ccttgaagca
     5461 gaagctgata ttgaaggtcg actaggaagt attccagata acaagttgcg ctagttggac
     5521 tttattacaa gcagttgcaa tcttcggacg attctgctgt cgctggttac cgtagaacta
     5581 ttcctaatgc ttgctcaatc cctatgagtt cacttgttgt tgtcggattt cccaattcgc
     5641 aagaagccga agaagtccga atcaaacttg ctcagctaca gacagaacat ttaattgccc
     5701 tagaagatgc tgttgttgtc agcaaggatt ccgagggcca tgtaaaactc aatcaagcga
     5761 ttaatttaac ggcaactggt gccgtcagcg gcggtttttg gggcaccctt gtcggcttga
     5821 tcttcctcaa tcccttggcg ggtgcggcga tcggtgcagc ggctggtgca ctcagcggtt
     5881 ccttgagcga cattggcatc aacgatgact ttctgcgcga ggtcggcgaa accctgcagg
     5941 tcggatcagc cgccctctgc ctcctcatcc gcgaagcaac ccccgatcgc atcctgaaag
     6001 agttgcgagt ctacgccaaa gacgcccgga ttctacaaac gaacttgagc tttgccgatg
     6061 aagaaaacct caaggcagcc ctagaaaaag ccaaagccga agcccaagcc ctgcgtctca
     6121 attaatctcc aacgcttcga tcgctcctga ttacttgagt tgatcaaggc tcagatcaag
     6181 agcgatcgct acttcaatac agcttacttt tggccgctat ctcaacagca gttcaatcct
     6241 aagaaactgc agaagcaggg attgagtaat ttgatggttg actgcagtcc ttatcagttg
     6301 cttctcaaca attatccaac cgcaattacg gacagagcac tggactagca ccactgctaa
     6361 aaccgcaaag ctcggacaaa tataaagaca acttaaagat tttctacgcc ctgcgacctc
     6421 accagaacag agccactatc atttttaaga aaacataaat tgatccgctg taagccgctg
     6481 ccagcacaaa gcttaccgcg tttttgctaa agcctagtcc gctggtttgt cctcccggaa
     6541 tgttgcccta tgaaaccccg catcctcgtg atctagcccc tggcgaaagg gggatgtgct
     6601 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgtt
//
