LOCUS       sequence-173608-        6541 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Golden Gate Level 1 cassette encoding a Nat selectable marker under 
            the FCP promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6541)
  AUTHORS   Hopes A, Nekrasov V, Kamoun S, Mock T
  TITLE     Editing of the urease gene by CRISPR-Cas in the diatom Thalassiosira
            pseudonana.
  JOURNAL   Plant Methods. 2016 Nov 24;12:49. doi: 10.1186/s13007-016-0148-0. 
            eCollection 2016.
  PUBMED    27904648
REFERENCE   2  (bases 1 to 6541)
  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: "Plant 
            Methods."; date: "2016-11-24"; volume: "12:49. doi"; pages: " 
            10.1186/s13007-016-0148-0. eCollection 2016"
COMMENT     Sequence Label: pICH47732:FCP:NAT
FEATURES             Location/Qualifiers
     source          1..6541
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     35..51
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     36..52
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1035..1604
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGFT
                     LREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNRR
                     LTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFTL
                     CGLDTALYDGTASDGEQALYMSMPCP"
     primer_bind     complement(2120..2136)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2120..2136)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2177..2197)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2177..2195)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    2244..2268
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2424..3572)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      complement(3908..4496)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3988..4007)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4667..5458,5459..5527))
                     /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     5290..5309
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5528..5632)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5719..6350
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    6442..6466
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 tctgtgaaga caatgccgaa ttcggatccg gagctcgagg tcgacggtat cgataagctt
       61 gcgctttttc cgagaactcc ccataagtca acggctccaa tcaagaatgt atccgacaac
      121 ggcgagcata gcaacacgtc cgtctttgga gtagaatcat catgttgtgg atgaatacac
      181 agatgaatga cattagaagc atgaacatgt tagagagtag gaggtagaga ttgatatggt
      241 agcattgcga tgtttgtttt tggtcagcat atgatgagtg gataccaata tgatgaaagt
      301 tgaatctcgc gtttgagctc agcggtacgt tattgatcga aagtagcctg atcaaaatcc
      361 ttggagagta caagaggatc aaagaatcca gtgggggcga taactccaag ctcgttctca
      421 aagaggcaat ggaggtagaa actcatccca gttgagaaga agtgaaggca gtggcggtgg
      481 cgaaagcaga ggcaacgagg acagacttcc tgtgggttga tgcaacgaat atttccagaa
      541 ggagaagttt agagagttga accgctacct acaatgacaa agtatcgtat cgattttgat
      601 gttggttggt tatgaattca aactgtaagt tggattgtga gaagatcaga agttgaacga
      661 acacatcttt ccgatcattc acctccacac tgcaacaaca cggtacttct tccgcggcag
      721 atctctgtcg ccattctctt gtcctgttgt tggctgtgag acgaggaaag caacgacaag
      781 tttcacaaaa gggagttcct ttaacgagat atgtttttta taaagagtcc caatagaaag
      841 acaaattgat tcctccgtgc aaacgcgcaa ataaacacca cgtccattat atccatatct
      901 ttcagagtat ccaacaagtg ttgaaggaca ggtagttgaa gtaacgtatc ttccccctcg
      961 actggatcca tcaacaaggc gaacaaatcc attcaacctc tcataaatta tctgatttac
     1021 caaaccgatc caaaatgacc actcttgacg acacggctta ccggtaccgc accagtgtcc
     1081 cgggggacgc cgaggccatc gaggcactgg atgggtcctt caccaccgac accgtattcc
     1141 gcgtcaccgc caccggggac ggcttcaccc tgcgggaggt gccggtggac ccgcccctga
     1201 ccaaggtgtt ccccgacgac gaatcggacg acgaatcgga cgacggggag gacggcgacc
     1261 cggactcccg gacgttcgtc gcgtacgggg acgacggcga cctggcgggc ttcgtggtcg
     1321 tctcgtactc cggctggaac cgccggctga ccgtcgagga catcgaggtc gccccggagc
     1381 accgggggca cggggtcggg cgcgcgttga tggggctcgc gacggagttc gcccgcgagc
     1441 ggggcgccgg gcacctctgg ctggaggtca ccaacgtcaa cgcaccggcg atccacgcgt
     1501 accggcggat ggggttcacc ctctgcggcc tggacaccgc cctgtacgac ggcaccgcct
     1561 cggacggcga gcaggcgctc tacatgagca tgccctgccc ctgagcggcc gcatactgga
     1621 ttggtgaatc aatgagccgt agcacaatgg ttacattcgg ctagctaaga tccaatggca
     1681 aggaccaagt gctggaactt gttttgcttt agcagatctt agcgtgagag gtatttgtcc
     1741 tctgtcagga gtagatagta gatgttcttt ttaaactaaa atgctaactg ttccgaattc
     1801 ctcatcgcag ctaatccgta catcaaaaga caaaatgcta ggtatgtgta ctacatctcc
     1861 tgttgctaga taagacatat gataggaaac acaccatcaa tagtcattgt agctttactt
     1921 atactacgca tttgcacttt cccctgagtg gcagaggcgc attgagaaaa tcgatctcaa
     1981 catagtttat gtagcatccc ctagatccat tactttaagt ctccttcgtc tttggtgtag
     2041 gcatgttgga cacaacgagg taaaacacaa cacaaacaat gtgtccagca aagtagtagc
     2101 tgctccagtt ctcccggggg atccactagt tctagagcgg ccggccgcca ccgcggtgga
     2161 gctccagctt ttgttccctt tagtgagggt taattgcgcg ctgcaattgt cttctgcacg
     2221 aagtggttta aactatcagt gtttgacagg atatattggc gggtaaacct aagagaaaag
     2281 agcgtttatt agaataatcg gatatttaaa agggcgtgaa aaggtttatc cgttcgtcca
     2341 tttgtatgtg catgccaacc acagggttcc ccagatcagg cgctggctgc tgaaccccca
     2401 gccggaactg accccacaag gccctagcgt ttgcaatgca ccaggtcatc attgacccag
     2461 gcgtgttcca ccaggccgct gcctcgcaac tcttcgcagg cttcgccgac ctgctcgcgc
     2521 cacttcttca cgcgggtgga atccgatccg cacatgaggc ggaaggtttc cagcttgagc
     2581 gggtacggct cccggtgcga gctgaaatag tcgaacatcc gtcgggccgt cggcgacagc
     2641 ttgcggtact tctcccatat gaatttcgtg tagtggtcgc cagcaaacag cacgacgatt
     2701 tcctcgtcga tcaggacctg gcaacgggac gttttcttgc cacggtccag gacgcggaag
     2761 cggtgcagca gcgacaccga ttccaggtgc ccaacgcggt cggacgtgaa gcccatcgcc
     2821 gtcgcctgta ggcgcgacag gcattcctcg gccttcgtgt aataccggcc attgatcgac
     2881 cagcccaggt cctggcaaag ctcgtagaac gtgaaggtga tcggctcgcc gataggggtg
     2941 cgcttcgcgt actccaacac ctgctgccac accagttcgt catcgtcggc ccgcagctcg
     3001 acgccggtgt aggtgatctt cacgtccttg ttgacgtgga aaatgacctt gttttgcagc
     3061 gcctcgcgcg ggattttctt gttgcgcgtg gtgaacaggg cagagcgggc cgtgtcgttt
     3121 ggcatcgctc gcatcgtgtc cggccacggc gcaatatcga acaaggaaag ctgcatttcc
     3181 ttgatctgct gcttcgtgtg tttcagcaac gcggcctgct tggcctcgct gacctgtttt
     3241 gccaggtcct cgccggcggt ttttcgcttc ttggtcgtca tagttcctcg cgtgtcgatg
     3301 gtcatcgact tcgccaaacc tgccgcctcc tgttcaagac gacgcgaacg ctccacggcg
     3361 gccgatggcg cgggcagggc agggggagcc agttgcacgc tgtcgcgctc gatcttggcc
     3421 gtagcttgct ggaccatcga gccgacggac tggaaggttt cgcggggcgc acgcatgacg
     3481 gtgcggcttg cgatggtttc ggcatcctcg gcggaaaacc ccgcgtcgat cagttcttgc
     3541 ctgtatgcct tccggtcaaa cgtccgattc attcaccctc cttgcgggat tgccccgact
     3601 cacgccgggg caatgtgccc ttattcctga tttgacccgc ctggtgcctt ggtgtccaga
     3661 taatccacct tatcggcaat gaagtcggtc ccgtagaccg tctggccgtc cttctcgtac
     3721 ttggtattcc gaatcttgcc ctgcacgaat accagcgacc ccttgcccaa atacttgccg
     3781 tgggcctcgg cctgagagcc aaaacacttg atgcggaaga agtcggtgcg ctcctgcttg
     3841 tcgccggcat cgttgcgcca catctaggat ctgccaggaa ccgtaaaaag gccgcgttgc
     3901 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3961 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4021 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4081 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4141 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4201 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4261 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4321 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4381 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4441 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4501 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4561 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4621 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4681 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4741 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4801 taccgcgaga accacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4861 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4921 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4981 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5041 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5101 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5161 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5221 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5281 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5341 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5401 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5461 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5521 tactcatact cttccttttt caatattatt gaagcattta tcaaggttat tgtctcatga
     5581 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacgaatt
     5641 ggccagcgct gccatttttg gggtgaggcc gttcgcggcc gaggggcgca gcccctgggg
     5701 ggatgggagg cccgcgttag cgggccggga gggttcgaga agggggggca ccccccttcg
     5761 gcgtgcgcgg tcacgcgcac agggcgcagc cctggttaaa aacaaggttt ataaatattg
     5821 gtttaaaagc aggttaaaag acaggttagc ggtggccgaa aaacgggcgg aaacccttgc
     5881 aaatgctgga ttttctgcct gtggacagcc cctcaaatgt caataggtgc gcccctcatc
     5941 tgtcagcact ctgcccctca agtgtcaagg atcgcgcccc tcatctgtca gtagtcgcgc
     6001 ccctcaagtg tcaataccgc agggcactta tccccaggct tgtccacatc atctgtggga
     6061 aactcgcgta aaatcaggcg ttttcgccga tttgcgaggc tggccagctc cacgtcgccg
     6121 gccgaaatcg agcctgcccc tcatctgtca acgccgcgcc gggtgagtcg gcccctcaag
     6181 tgtcaacgtc cgcccctcat ctgtcagtga gggccaagtt ttccgcgagg tatccacaac
     6241 gccggcggcc gcggtgtctc gcacacggct tcgacggcgt ttctggcgcg tttgcagggc
     6301 catagacggc cgccagccca gcggcgaggg caaccagccc ggtgagcgtc gcaaaggaga
     6361 tcctgatctg actgatgggc tgcctgtatc gagtggtgat tttgtgccga gctgccggtc
     6421 ggggagctgt tggctggctg gtggcaggat atattgtggt gtaaacaaat tgacgcttag
     6481 acaacttaat aacacattgc ggacgttttt aatgtactgg ggtggatgca gtgggcccca
     6541 c
//
