LOCUS       sequence-217260-       10018 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 2 cloning vector, for golden gate cloning for diatoms. All the
            Agrobacterium T-DNA delivery were deleted..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10018)
  AUTHORS   Graff van Creveld S, Coesel SN, Blaskowski S, Groussman RD, Schatz 
            MJ, Armbrust EV
  TITLE     Divergent functions of two clades of flavodoxin in diatoms mitigate 
            oxidative stress and iron limitation.
  JOURNAL   Elife. 2023 Jun 6;12:e84392. doi: 10.7554/eLife.84392.
  PUBMED    37278403
REFERENCE   2  (bases 1 to 10018)
  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: "2023-06-6"; volume: "12:e84392. doi"; pages: " 
            10.7554/eLife.84392"
COMMENT     Sequence Label: pAGM4723-Del
FEATURES             Location/Qualifiers
     source          1..10018
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      4085..4094
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        6101..6205
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6206..7000
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      7171..7759
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7660..7679
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7895..7919
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 gttcatacct atcgcgacct gttaagccag caccctgccg cggttaagca gtccaacaaa
       61 ctgcagacta agcgcatgag taactctctg tttgtgctct attttggttt gaatcaccat
      121 catgatcagc tcgcgcatca cacggtttgt ttcggcccgc gttaccgcga gctgattgac
      181 gaaattttta atcatgatgg cctcgcagag gacttctcac tttatctgca cgcgccctgt
      241 gtcacggatt cgtcactggc gcctgaaggt tgcggcagtt actatgtgtt ggcgccggtg
      301 ccgcatttag gcaccgcgaa cctcgactgg acggttgagg ggccaaaact acgcgaccgt
      361 atttttgcgt accttgagca gcattacatg cctggcttac ggagtcagct ggtcacgcac
      421 cggatgttta cgccgtttga ttttcgcgac cagcttaatg cctatcatgg ctcagccttt
      481 tctgtggagc ccgttcttac ccagagcgcc tggtttcggc cgcataaccg cgataaaacc
      541 attactaatc tctacctggt cggcgcaggc acgcatcccg gcgcaggcat tcctggcgtc
      601 atcggctcgg caaaagcgac agcaggtttg atgctggagg atctgatatg agcttttacc
      661 caggagacag agaatgcaac cgcattatga tctgattctc gtgggggctg gactcgcgaa
      721 tggccttatc gccctgcgac ttcagcagca gcaacctgat atgcgtattt tgcttatcga
      781 cgccgcaccc caggcgggcg ggaatcatac gtggtcattt caccacgatg atttgactga
      841 gagccaacat cgttggatag ctccgctggt ggttcatcac tggcccgact atcaggtacg
      901 ctttcccaca cgccgtcgta agctgaacag cggctacttt tgtattactt ctcagcgttt
      961 cgctgaggtt ttacagcgac agtttggccc gcacttgtgg atggataccg cggtcgcaga
     1021 ggttaatgcg gaatctgttc ggttgaaaaa gggtcaggtt atcggtgccc gcgcggtgat
     1081 tgacgggcgg ggttatgcgg caaattcagc actgagcgtg ggcttccagg cgtttattgg
     1141 ccaggaatgg cgattgagtc acccgcatgg tttatcatct ccgattatca tggatgccac
     1201 ggtcgatcag caaaatggtt atcgcttcgt gtacagcctg ccgctctcgc cgaccagatt
     1261 gttaattgaa gatacgcact atattgataa tgcgacatta gatcctgaat gcgcgcggca
     1321 aaatatttgc gactatgccg cgcaacaggg ttggcagctt cagacactgc tgcgagaaga
     1381 acagggcgcc ttacccatta ctctgtcggg caatgccgac gcattctggc agcagcgccc
     1441 cctggcctgt agtggattac gtgccggtct gttccatcct accaccggct attcactgcc
     1501 gctggcggtt gccgtggccg accgcctgag tgcacttgat gtctttacgt cggcctcaat
     1561 tcaccatgcc attacgcatt ttgcccgcga gcgctggcag cagcagggct ttttccgcat
     1621 gctgaatcgc atgctgtttt tagccggacc cgccgattca cgctggcggg ttatgcagcg
     1681 tttttatggt ttacctgaag atttaattgc ccgtttttat gcgggaaaac tcacgctgac
     1741 cgatcggcta cgtattctga gcggcaagcc gcctgttccg gtattagcag cattgcaagc
     1801 cattatgacg actcatcgtt aagcttcaga ggaggaatta agtaatgagt tttgatattg
     1861 ccaaataccc gaccctggca ctggtcgact ccacccagga gttacgactg ttgccgaaag
     1921 agagtttacc gaaactctgc gacgaactgc gccgctattt actcgacagc gtgagccgtt
     1981 ccagcgggca cttcgcctcc gggctgggca cggtcgaact gaccgtggcg ctgcactatg
     2041 tctacaacac cccgtttgac caattgattt gggatgtggg gcatcaggct tatccgcata
     2101 aaattttgac cggacgccgc gacaaaatcg gcaccatccg tcagaaaggc ggtctgcacc
     2161 cgttcccgtg gcgcggcgaa agcgaatatg acgtattaag cgtcgggcat tcatcaacct
     2221 ccattagtgc cggaattggt attgcggttg ctgccgaaaa agaaggcaaa aatcgccgca
     2281 ccgtctgtgt cattggcgat ggcgcgatta ccgcaggcat ggcgtttgaa gcgatgaatc
     2341 acgcgggcga tatccgtcct gatatgctgg tgattctcaa cgacaatgaa atgtcgattt
     2401 ccgaaaatgt cggcgcgctc aacaaccatc tggcacagct gctttccggt aagctttact
     2461 cttcactgcg cgaaggcggg aaaaaagttt tctctggcgt gccgccaatt aaagagctgc
     2521 tcaaacgcac cgaagaacat attaaaggca tggtagtgcc tggcacgttg tttgaggagc
     2581 tgggctttaa ctacatcggc ccggtggacg gtcatgatgt gctggggctt atcaccacgc
     2641 taaagaacat gcgcgacctg aaaggcccgc agttcctgca tatcatgacc aaaaaaggtc
     2701 gtggttatga accggcagaa aaagacccga tcactttcca tgccgtgcta aatttgatcc
     2761 ctccagcggt tgtttgccga aaagtagcgg cggtttgccg agctattcaa aaatctttgg
     2821 cgactggttg tgcgaaacgg cagcgaaaga caacaagctg atggcgatta ctccggcgat
     2881 gcgtgaaggt tccggcatgg tcgagttttc acgtaaattc ccggatcgct acttcgacgt
     2941 ggcaattgcc gagcaacatg cggtgacctt tgctgcgggt ctggcgattg gtgggtacaa
     3001 acccattgtc gcgatttact ccactttcct gcaacgcgcc tatgatcagg tgctgcatga
     3061 cgtggcgatt caaaagcttc cggtcctgtt cgccatcgac cgcgcgggca ttgttggtgc
     3121 tgacggtcaa acccatcagg gtgcttttga tctctcttac ctgcgctgca taccggaaat
     3181 ggtcattatg accccgagcg atgaaaacga atgtcgccag atgctctata ccggctatca
     3241 ctataacgat ggcccgtcag cggtgcgcta cccgcgtggc aacgcggtcg gcgtggaact
     3301 gacgccgctg gaaaaactac caattggcaa aggcattgtg aagcgtcgtg gcgagaaact
     3361 ggcgatcctt aactttggta cgctgatgcc agaagcggcg aaagtcgccg aatcgctgaa
     3421 cgccacgctg gtcgatatgc gttttgtgaa accgcttgat gaagcgttaa ttctggaaat
     3481 ggccgccagc catgaagcgc tggtcaccgt agaagaaaac gccattatgg gcggcgcagg
     3541 cagcggcgtg aacgaagtgc tgatggccca tcgtaaacca gtacccgtgc tgaacattgg
     3601 cctgccggac ttctttattc cgcaaggaac tcaggaagaa atgcgcgccg aactcggcct
     3661 cgatgccgct ggtatggaag ccaaaatcaa ggcctggctg gcataagctt tgtgtgagga
     3721 gatgtcgaat gagcgcacat gccctgccca aggcagatct gaccgccacc agcctgatcg
     3781 tatcgggcgg catcatcgcc gcttggctgg ccctgcatgt gcatgcgctg tggtttctgg
     3841 acgcagcggc gcatcccatc ctggcgatcg caaatttcct ggggctgacc tggctgtcgg
     3901 tcggattgtt catcatcgcg catgacgcga tgcacgggtc ggtggtgccg gggcgtccgc
     3961 gcgccaatgc ggcgatgggc cagcttgtcc tgtggctgta tgccggattt tcgtggcgca
     4021 agatgatcgt caagcacatg gcccatcacc gccatgccgg aaccgacgac gaccccgatt
     4081 tcgaccatgg cggcccggtc cgctggtacg cccgcttcat cggcacctat ttcggctggc
     4141 gcgaggggct gcttctgccc gtcatcgtga cggtctatgc gctgatcctt ggggatcgct
     4201 ggatgtacgt ggtattctgg ccgctgccgt cgatcctggc gtcgatccag ctgttcgtgt
     4261 tcggcacctg gctgccgcac cgccccggcc acgacgcgtt cccggaccgc cacaatgcgc
     4321 ggtcgtcgcg gatcagcgac cccgtgtcgc tgctgacctg ttttcacttt ggcggttatc
     4381 atcacgaaca ccacctgcac ccgacggtgc cgtggtggcg cctgcccagc acccgcacca
     4441 agggcgacac cgcataagct tgagctgagg agattttgaa tgcaaacgga acacgtcatt
     4501 ttattgaatg cacagggagt tcccacgggt acgctggaaa agtatgccgc acacacggca
     4561 gacacccgct tacatctcgc gttctccagt tggctgttta atgccaaagg acaattatta
     4621 gttacccgcc gcgcactgag caaaaaagca tggcctggcg tgtggactaa ctcggtttgt
     4681 gggcacccac aactgggaga aagcaacgag gacgcagtga tccgccgttg ccgttatgag
     4741 cttggcgtgg aaattacgcc tcctgaatct atctatcctg actttcgcta ccgcgccacc
     4801 gatccgagtg gcattgtgga aaatgaagtg tgtccggtat ttgccgcacg caccactagt
     4861 gcgttacaga tcaatgatga tgaagtgatg gattatcaat ggtgtgattt agcagatgta
     4921 ttacacggta ttgatgcaac gccgtgggcg ttcagtccgt ggatggtgat gcaggcgaca
     4981 aatcgcgaag ccagaaaacg attatctgca tttacccagc ttaaataagc tttgcctcag
     5041 gatttttgaa atgacggtct gcgcaaaaaa acacgttcat ctcactcgcg atgctgcgga
     5101 gcagttactg gctgatattg atcgacgcct tgatcagtta ttgcccgtgg agggagaacg
     5161 ggatgttgtg ggtgccgcga tgcgtgaagg tgcgctggca ccgggaaaac gtattcgccc
     5221 catgttgctg ttgctgaccg cccgcgatct gggttgcgct gtcagccatg acggattact
     5281 ggatttggcc tgtgcggtgg aaatggtcca cgcggcttcg ctgatccttg acgatatgcc
     5341 ctgcatggac gatgcgaagc tgcggcgcgg acgccctacc attcattctc attacggaga
     5401 gcatgtggca atactggcgg cggttgcctt gctgagtaaa gcctttggcg taattgccga
     5461 tgcagatggc ctcacgccgc tggcaaaaaa tcgggcggtt tctgaactgt caaacgccat
     5521 cggcatgcaa ggattggttc agggtcagtt caaggatctg tctgaagggg ataagccgcg
     5581 cagcgctgaa gctattttga tgacgaatca ctttaaaacc agcacgctgt tttgtgcctc
     5641 gatgcagatg gcctcgattg ttgcgaatgc ctccagcgaa gcgcgtgatt gcctgcatcg
     5701 tttttcactt gatcttggtc aggcatttca actgctggac gatttgaccg atggcatgac
     5761 cgacaccggt aaggatagca atcaggacgc cggtaaatcg acgctggtca atctgttagg
     5821 cccgagggcg gttgaagaac gtctgagaca acatcttcag cttgccagtg agcatctctc
     5881 tgcggcctgc caacacgggc acgccactca acattttatt caggcctggt ttgacaaaaa
     5941 actcgctgcc gtcagttaag cttgcaagaa ttcaagcttg gagtacgtta attagttgtc
     6001 tgttttaagc cccgtgctct ccgccggggc ttttttatta aaaaccactc aagtggaaga
     6061 caagggacac gaagtgatcc ccacatcaag gctccgagtg cgcggaaccc ctatttgttt
     6121 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     6181 tcaataatat tgaaaaagga agagtatggc taaaatgaga atatcaccgg aattgaaaaa
     6241 actgatcgaa aaataccgct gcgtaaaaga tacggaagga atgtctcctg ctaaggtata
     6301 taagctggtg ggagaaaatg aaaacctata tttaaaaatg acggacagcc ggtataaagg
     6361 gaccacctat gatgtggaac gggaaaagga catgatgcta tggctggaag gaaagctgcc
     6421 tgttccaaag gtcctgcact ttgaacggca tgatggctgg agcaatctgc tcatgagtga
     6481 ggccgatggc gtcctttgct cggaagagta tgaagatgaa caaagccctg aaaagattat
     6541 cgagctgtat gcggagtgca tcaggctctt tcactccatc gacatatcgg attgtcccta
     6601 tacgaatagc ttagacagcc gcttagccga attggattac ttactgaata acgatctggc
     6661 cgatgtggat tgcgaaaact gggaagagga cactccattt aaagatccgc gcgagctgta
     6721 tgatttttta aagacggaaa agcccgaaga ggaacttgtc ttttcccacg gcgacctggg
     6781 agacagcaac atctttgtga aagatggcaa agtaagtggc tttattgatc ttgggagaag
     6841 cggcagggcg gacaagtggt atgacattgc cttctgcgtc cggtcgctca gggaggatat
     6901 cggggaagaa cagtatgtcg agctattttt tgacttactg gggatcaagc ctgattggga
     6961 gaaaataaaa tattatattt tactggatga attgttttag ctgtcagacc aagtttactc
     7021 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     7081 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     7141 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     7201 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     7261 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     7321 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     7381 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     7441 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     7501 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     7561 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     7621 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     7681 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     7741 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tgctcggatc
     7801 tgttggaccg gacagtagtc atggttgatg ggctgcctgt atcgagtggt gattttgtgc
     7861 cgagctgccg gtcggggagc tgttggctgg ctggtggcag gatatattgt ggtgtaaaca
     7921 aattgacgct tagacaactt aataacacat tgcggacgtt tttaatgtac tggggttgaa
     7981 cactctgtgc cttgtcttcc agtggtatgg ggtaccgcac ggtctgccaa tccgacggag
     8041 gtttatgaat tttccacctt ttccacaagc tcaactagta ttaacgatgt ggatttagca
     8101 aaaaaaacct gtaaccctaa atgtaaaata acgggtaagc ctgccaacca tgttatggca
     8161 gattaagcgt ctttttgaag ggcaccgcat ctttcgcgtt gccgtaaatg tatccgttta
     8221 gcaataagga aattcttaat gaataatccg tcgttactca atcatgcggt cgaaacgatg
     8281 gcagttggct cgaaaagttt tgcgacagcc tcaaagttat ttgatgcaaa aacccggcgc
     8341 agcgtactga tgctctacgc ctggtgccgc cattgtgacg atgttattga cgatcagacg
     8401 ctgggctttc aggcccggca gcctgcctta caaacgcccg aacaacgtct gatgcaactt
     8461 gagatgaaaa cgcgccaggc ctatgcagga tcgcagatgc acgaaccggc gtttgcggct
     8521 tttcgggaag tggctatggc tcatgatatc gccccggctt acgcgtttga tcatctggaa
     8581 ggcttcgcca tggatgtacg cgaagcgcaa tacagccaac tggatgatac gctgcgctat
     8641 tgctatcacg ttgcaggcgt tgtcggcttg atgatggcgc aaatcatggg cgtgcgggat
     8701 aacgccacgc tggaccgcgc ctgtgacctt gggctggcat ttcagttgac caatattgct
     8761 cgcgatattg tggacgatgc gcatgcgggc cgctgttatc tgccggcaag ctggctggag
     8821 catgaaggtc tgaacaaaga gaattatgcg gcacctgaaa accgtcaggc gctgagccgt
     8881 atcgcccgtc gtttggtgca ggaagcagaa ccttactatt tgtctgccac agccggcctg
     8941 gcagggttgc ccctgcgttc cgcctgggca atcgctacgg cgaagcaggt ttaccggaaa
     9001 ataggtgtca aagttgaaca ggccggtcag caagcctggg atcagcggca gtcaacgacc
     9061 acgcccgaaa aattaacgct gctgctggcc gcctctggtc aggcccttac ttcccggatg
     9121 cgggctcatc ctccccgccc tgcgcatctc tggcagcgcc cgctctaggc ttactataag
     9181 gatgtttgca atgaaaccaa ctacggtaat tggtgcaggc ttcggtggcc tggcactggc
     9241 aattcgtcta caagctgcgg ggatccccgt cttactgctt gaacaacgtg ataaacccgg
     9301 cggtcgggct tatgtctacg aggatcaggg gtttaccttt gatgcaggcc cgacggttat
     9361 caccgatccc agtgccattg aagaactgtt tgcactggca ggaaaacagt taaaagagta
     9421 tgtcgaactg ctgccggtta cgccgtttta ccgcctgtgt tgggagtcag ggaaggtctt
     9481 taattacgat aacgatcaaa cccggctcga agcgcagatt cagcagttta atccccgcga
     9541 tgtcgaaggt tatcgtcagt ttctggacta ttcacgcgcg gtgtttaaag aaggctatct
     9601 aaagctcggt actgtccctt ttttatcgtt cagagacatg cttcgcgccg cacctcaact
     9661 ggcgaaactg caggcatgga gaagcgttta cagtaaggtt gccagttaca tcgaagatga
     9721 acatctgcgc caggcgtttt ctttccactc gctgttggtg ggcggcaatc ccttcgccac
     9781 ctcatccatt tatacgttga tacacgcgct ggagcgtgag tggggcgtct ggtttccgcg
     9841 tggcggcacc ggcgcattag ttcaggggat gataaagctg tttcaggatc tgggtggcga
     9901 agtcgtgtta aacgccagag tcagccatat ggaaacgaca ggaaacaaga ttgaagccgt
     9961 gcatttagag gacggtcgca ggttcctgac gcaagccgtc gcgtcaaatg cagatgtg
//
