pSPYCE35S-bk1-3Flag-bk2

LOCUS       pSPYCE35S-bk1-3Flag-bk2               3951 bp DNA     circular SYN 31-OCT-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3951)
  AUTHORS   Youbio
  TITLE     Direct Submission
  JOURNAL   Exported Sep 22, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..3951
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     26..42
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        566..911
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     852..871
                     /label=YNEF
     promoter        920..922
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     CDS             971..1042
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDKM"
     misc_feature    1049..1069
                     /label=同源臂
     misc_feature    1082..1102
                     /label=同源臂
     terminator      1116..1368
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(1377..1393)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1867..1971
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1972..2832
                     /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"
     rep_origin      3003..3591
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctt
       61 gcatgcctgc aggtccccag attagccttt tcaatttcag aaagaatgct aacccacaga
      121 tggttagaga ggcttacgca gcaggtctca tcaagacgat ctacccgagc aataatctcc
      181 aggaaatcaa ataccttccc aagaaggtta aagatgcagt caaaagattc aggactaact
      241 gcatcaagaa cacagagaaa gatatatttc tcaagatcag aagtactatt ccagtatgga
      301 cgattcaagg cttgcttcac aaaccaaggc aagtaataga gattggagtc tctaaaaagg
      361 tagttcccac tgaatcaaag gccatggagt caaagattca aatagaggac ctaacagaac
      421 tcgccgtaaa gactggcgaa cagttcatac agagtctctt acgactcaat gacaagaaga
      481 aaatcttcgt caacatggtg gagcacgaca cacttgtcta ctccaaaaat atcaaagata
      541 cagtctcaga agaccaaagg gcaattgaga cttttcaaca aagggtaata tccggaaacc
      601 tcctcggatt ccattgccca gctatctgtc actttattgt gaagatagtg gaaaaggaag
      661 gtggctccta caaatgccat cattgcgata aaggaaaggc catcgttgaa gatgcctctg
      721 ccgacagtgg tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagacg
      781 ttccaaccac gtcttcaaag caagtggatt gatgtgatat ctccactgac gtaagggatg
      841 acgcacaatc ccactatcct tcgcaagacc cttcctctat ataaggaagt tcatttcatt
      901 tggagagaac acgggggacc aggtcgactc tcgaggatcc ggtacccggt atcgatttaa
      961 aaggcctatg gactacaagg atgacgacga caaggactac aaggatgacg acgacaagga
     1021 ctacaaggac gacgacaaga tgaggcctca ggtcgactct cgaggatcaa ctagtgagct
     1081 cagtacccgg tatcgattta aataagaatt tccccgatcg ttcaaacatt tggcaataaa
     1141 gtttcttaag attgaatcct gttgccggtc ttgcgatgat tatcatataa tttctgttga
     1201 attacgttaa gcatgtaata attaacatgt aatgcatgac gttatttatg agatgggttt
     1261 ttatgattag agtcccgcaa ttatacattt aatacgcgat agaaaacaaa atatagcgcg
     1321 caaactagga taaattatcg cgcgcggtgt catctatgtt actagatcgg gaattcactg
     1381 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     1441 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     1501 tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg
     1561 catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc
     1621 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     1681 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     1741 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     1801 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     1861 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     1921 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     1981 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     2041 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     2101 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     2161 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     2221 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     2281 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2341 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     2401 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     2461 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     2521 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     2581 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     2641 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     2701 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     2761 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     2821 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     2881 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     2941 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3001 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3061 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3121 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     3181 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3241 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3301 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3361 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3421 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3481 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3541 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3601 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3661 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     3721 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     3781 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     3841 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     3901 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg a
//