LOCUS       sequence-225201-        6693 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FAST-Red selectable marker Golden Gate Level 1 Position 1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6693)
  AUTHORS   Castel B, Tomlinson L, Locci F, Yang Y, Jones JDG
  TITLE     Optimization of T-DNA architecture for Cas9-mediated mutagenesis in 
            Arabidopsis.
  JOURNAL   PLoS One. 2019 Jan 9;14(1):e0204778. doi: 
            10.1371/journal.pone.0204778. eCollection 2019.
  PUBMED    30625150
REFERENCE   2  (bases 1 to 6693)
  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: "PLoS One.";
            date: "2019-01-9"; volume: "14(1):e0204778. doi"; pages: " 
            10.1371/journal.pone.0204778. eCollection 2019"
COMMENT     Sequence Label: pICSL11015
FEATURES             Location/Qualifiers
     source          1..6693
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1213..1908
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSDMALK
                     LVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHK"
     misc_feature    2396..2420
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2576..3724)
                     /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(4060..4648)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4140..4159)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4819..5610,5611..5679))
                     /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     5442..5461
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5680..5784)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5871..6502
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    6594..6618
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 tctgtgaaga caatgccgaa ttcggatccg gagttctaga atgtcgcgga acaaatttta
       61 aaactaaatc ctaaattttt ctaattttgt tgccaatagt ggatatgtgg gccgtataga
      121 aggaatctat tgaaggccca aacccatact gacgagccca aaggttcgtt ttgcgtttta
      181 tgtttcggtt cgatgccaac gccacattct gagctaggca aaaaacaaac gtgtctttga
      241 atagactcct ctcgttaaca catgcagcgg ctgcatggtg acgccattaa cacgtggcct
      301 acaattgcat gatgtctcca ttgacacgtg acttctcgtc tcctttctta atatatctaa
      361 caaacactcc tacctcttcc aaaatatata cacatctttt tgatcaatct ctcattcaaa
      421 atctcattct ctctagtaaa caagaacaaa aaaatggcgg atacagctag aggaacccat
      481 cacgatatca tcggcagaga tcagtacccg atgatgggcc gagatcgtga ccagtaccag
      541 atgtccggac gaggatctga ctactccaag tctaggcaga ttgctaaagc tgcaactgct
      601 gtcacagctg gtggttccct ccttgttctc tccagcctta cccttgttgg aactgtcata
      661 gctttgactg ttgcaacacc tctgctcgtt atcttcagcc caatccttgt cccggctctc
      721 atcacagttg cactcctcat caccggtttt ctttcctctg gagggtttgg cattgccgct
      781 ataaccgttt tctcttggat ttacaagtaa gcacacattt atcatcttac ttcataattt
      841 tgtgcaatat gtgcatgcat gtgttgagcc agtagctttg gatcaatttt tttggtcgaa
      901 taacaaatgt aacaataaga aattgcaaat tctagggaac atttggttaa ctaaatacga
      961 aatttgacct agctagcttg aatgtgtctg tgtatatcat ctatataggt aaaatgcttg
     1021 gtatgatacc tattgattgt gaataggtac gcaacgggag agcacccaca gggatcagac
     1081 aagttggaca gtgcaaggat gaagttggga agcaaagctc aggatctgaa agacagagct
     1141 cagtactacg gacagcaaca tactggtggg gaacatgacc gtgaccgtac tcgtggtggc
     1201 cagcacacta ctatgagcga gctgattaag gagaacatgc acatgaagct gtacatggag
     1261 ggcaccgtga acaaccacca cttcaagtgc acatccgagg gcgaaggcaa gccctacgag
     1321 ggcacccaga ccatgagaat caaggtggtc gagggcggcc ctctcccctt cgccttcgac
     1381 atcctggcta ccagcttcat gtacggcagc agaaccttca tcaaccacac ccagggcatc
     1441 cccgacttct ttaagcagtc cttccctgag ggcttcacat gggagagagt caccacatac
     1501 gaagatgggg gcgtgctgac cgctacccag gacaccagcc tccaggacgg ctgcctcatc
     1561 tacaacgtca agatcagagg ggtgaacttc ccatccaacg gccctgtgat gcagaagaaa
     1621 acactcggct gggaggccaa caccgagatg ctgtaccccg ctgacggcgg cctggaaggc
     1681 agaagcgaca tggccctgaa gctcgtgggc gggggccacc tgatctgcaa cttcaagacc
     1741 acatacagat ccaagaaacc cgctaagaac ctcaagatgc ccggcgtcta ctatgtggac
     1801 cacagactgg aaagaatcaa ggaggccgac aaagaaacct acgtcgagca gcacgaggtg
     1861 gctgtggcca gatactgcga cctccctagc aaactggggc acaagtgagc ttaccccact
     1921 gatgtcatcg tcatagtcca ataactccaa tgtcggggag ttagtttatg aggaataaag
     1981 tgtttagaat ttgatcaggg ggagataata aaagccgagt ttgaatcttt ttgttataag
     2041 taatgtttat gtgtgtttct atatgttgtc aaatggtacc atgttttttt tcctctcttt
     2101 ttgtaacttg caagtgttgt gttgtacttt atttggcttc tttgtaagtt ggtaacggtg
     2161 gtctatatat ggaaaaggtc ttgttttgtt aaacttatgt tagttaactg gattcgtctt
     2221 taaccacaaa aagttttcaa taagctacaa atttagacac gcaagccgat gcagtcatta
     2281 gtacatatat ttattgcaag tgattacatg gcaacccaaa cttcaaaaac agtaggttgc
     2341 tccatttagt cgctgcaatt gtcttctgca cgaagtggtt taaactatca gtgtttgaca
     2401 ggatatattg gcgggtaaac ctaagagaaa agagcgttta ttagaataat cggatattta
     2461 aaagggcgtg aaaaggttta tccgttcgtc catttgtatg tgcatgccaa ccacagggtt
     2521 ccccagatca ggcgctggct gctgaacccc cagccggaac tgaccccaca aggccctagc
     2581 gtttgcaatg caccaggtca tcattgaccc aggcgtgttc caccaggccg ctgcctcgca
     2641 actcttcgca ggcttcgccg acctgctcgc gccacttctt cacgcgggtg gaatccgatc
     2701 cgcacatgag gcggaaggtt tccagcttga gcgggtacgg ctcccggtgc gagctgaaat
     2761 agtcgaacat ccgtcgggcc gtcggcgaca gcttgcggta cttctcccat atgaatttcg
     2821 tgtagtggtc gccagcaaac agcacgacga tttcctcgtc gatcaggacc tggcaacggg
     2881 acgttttctt gccacggtcc aggacgcgga agcggtgcag cagcgacacc gattccaggt
     2941 gcccaacgcg gtcggacgtg aagcccatcg ccgtcgcctg taggcgcgac aggcattcct
     3001 cggccttcgt gtaataccgg ccattgatcg accagcccag gtcctggcaa agctcgtaga
     3061 acgtgaaggt gatcggctcg ccgatagggg tgcgcttcgc gtactccaac acctgctgcc
     3121 acaccagttc gtcatcgtcg gcccgcagct cgacgccggt gtaggtgatc ttcacgtcct
     3181 tgttgacgtg gaaaatgacc ttgttttgca gcgcctcgcg cgggattttc ttgttgcgcg
     3241 tggtgaacag ggcagagcgg gccgtgtcgt ttggcatcgc tcgcatcgtg tccggccacg
     3301 gcgcaatatc gaacaaggaa agctgcattt ccttgatctg ctgcttcgtg tgtttcagca
     3361 acgcggcctg cttggcctcg ctgacctgtt ttgccaggtc ctcgccggcg gtttttcgct
     3421 tcttggtcgt catagttcct cgcgtgtcga tggtcatcga cttcgccaaa cctgccgcct
     3481 cctgttcaag acgacgcgaa cgctccacgg cggccgatgg cgcgggcagg gcagggggag
     3541 ccagttgcac gctgtcgcgc tcgatcttgg ccgtagcttg ctggaccatc gagccgacgg
     3601 actggaaggt ttcgcggggc gcacgcatga cggtgcggct tgcgatggtt tcggcatcct
     3661 cggcggaaaa ccccgcgtcg atcagttctt gcctgtatgc cttccggtca aacgtccgat
     3721 tcattcaccc tccttgcggg attgccccga ctcacgccgg ggcaatgtgc ccttattcct
     3781 gatttgaccc gcctggtgcc ttggtgtcca gataatccac cttatcggca atgaagtcgg
     3841 tcccgtagac cgtctggccg tccttctcgt acttggtatt ccgaatcttg ccctgcacga
     3901 ataccagcga ccccttgccc aaatacttgc cgtgggcctc ggcctgagag ccaaaacact
     3961 tgatgcggaa gaagtcggtg cgctcctgct tgtcgccggc atcgttgcgc cacatctagg
     4021 atctgccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4081 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4141 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4201 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4261 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4321 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4381 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4441 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4501 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4561 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4621 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4681 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4741 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4801 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4861 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4921 gggcttacca tctggcccca gtgctgcaat gataccgcga gaaccacgct caccggctcc
     4981 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5041 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5101 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5161 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5221 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5281 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5341 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5401 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5461 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5521 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5581 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5641 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5701 ttgaagcatt tatcaaggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5761 aaataaacaa ataggggttc cgcgcacgaa ttggccagcg ctgccatttt tggggtgagg
     5821 ccgttcgcgg ccgaggggcg cagcccctgg ggggatggga ggcccgcgtt agcgggccgg
     5881 gagggttcga gaaggggggg cacccccctt cggcgtgcgc ggtcacgcgc acagggcgca
     5941 gccctggtta aaaacaaggt ttataaatat tggtttaaaa gcaggttaaa agacaggtta
     6001 gcggtggccg aaaaacgggc ggaaaccctt gcaaatgctg gattttctgc ctgtggacag
     6061 cccctcaaat gtcaataggt gcgcccctca tctgtcagca ctctgcccct caagtgtcaa
     6121 ggatcgcgcc cctcatctgt cagtagtcgc gcccctcaag tgtcaatacc gcagggcact
     6181 tatccccagg cttgtccaca tcatctgtgg gaaactcgcg taaaatcagg cgttttcgcc
     6241 gatttgcgag gctggccagc tccacgtcgc cggccgaaat cgagcctgcc cctcatctgt
     6301 caacgccgcg ccgggtgagt cggcccctca agtgtcaacg tccgcccctc atctgtcagt
     6361 gagggccaag ttttccgcga ggtatccaca acgccggcgg ccgcggtgtc tcgcacacgg
     6421 cttcgacggc gtttctggcg cgtttgcagg gccatagacg gccgccagcc cagcggcgag
     6481 ggcaaccagc ccggtgagcg tcgcaaagga gatcctgatc tgactgatgg gctgcctgta
     6541 tcgagtggtg attttgtgcc gagctgccgg tcggggagct gttggctggc tggtggcagg
     6601 atatattgtg gtgtaaacaa attgacgctt agacaactta ataacacatt gcggacgttt
     6661 ttaatgtact ggggtggatg cagtgggccc cac
//
