pSPYCE35S-bk1-5HA-bk2

LOCUS       pSPYCE35S-bk1-5HA-bk2             4014 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 4014)
  AUTHORS   Youbio
  TITLE     Direct Submission
  JOURNAL   Exported Sep 22, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..4014
                     /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..1051
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             998..1078
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             1025..1105
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     misc_feature    1112..1132
                     /label=同源臂
     misc_feature    1145..1165
                     /label=同源臂
     terminator      1179..1431
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(1440..1456)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1930..2034
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2035..2895
                     /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      3066..3654
                     /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 tacccatacg atgttccaga ttacgcttac ccatacgatg ttccagatta
     1021 cgcttaccca tacgatgttc cagattacgc ttacccatac gatgttccag attacgctta
     1081 cccatacgat gttccagatt acgctaggcc tcaggtcgac tctcgaggat caactagtga
     1141 gctcagtacc cggtatcgat ttaaataaga atttccccga tcgttcaaac atttggcaat
     1201 aaagtttctt aagattgaat cctgttgccg gtcttgcgat gattatcata taatttctgt
     1261 tgaattacgt taagcatgta ataattaaca tgtaatgcat gacgttattt atgagatggg
     1321 tttttatgat tagagtcccg caattataca tttaatacgc gatagaaaac aaaatatagc
     1381 gcgcaaacta ggataaatta tcgcgcgcgg tgtcatctat gttactagat cgggaattca
     1441 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     1501 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     1561 ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt
     1621 acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat
     1681 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     1741 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     1801 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     1861 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     1921 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     1981 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     2041 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     2101 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     2161 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     2221 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     2281 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     2341 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     2401 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     2461 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     2521 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     2581 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     2641 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     2701 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     2761 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     2821 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     2881 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     2941 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     3001 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3061 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3121 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3181 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     3241 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3301 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3361 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3421 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3481 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3541 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3601 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3661 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     3721 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     3781 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     3841 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     3901 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     3961 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tgga
//