LOCUS       sequence-405376-        4981 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pNpuDnaE(C)_HygR(53-341)_3HA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4981)
  AUTHORS   Yuan G, Lu H, De K, Hassan MM, Liu Y, Islam MT, Muchero W, Tuskan 
            GA, Yang X
  TITLE     Split selectable marker systems utilizing inteins facilitate gene 
            stacking in plants.
  JOURNAL   Commun Biol. 2023 May 26;6(1):567. doi: 10.1038/s42003-023-04950-8.
  PUBMED    37237044
REFERENCE   2  (bases 1 to 4981)
  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: "Commun 
            Biol."; date: "2023-05-26"; volume: "6(1):567. doi"; pages: " 
            10.1038/s42003-023-04950-8"
COMMENT     Sequence Label: pAXY0019
FEATURES             Location/Qualifiers
     source          1..4981
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(447..1238,1239..1307))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGASKGGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1070..1089
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1308..1412)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1438..1893)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1575..1596)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1787..1806
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(2779..2790)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        2840..2858
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2840..2857
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             3868..3948
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     polyA_signal    3981..4115
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4005..4024)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4059..4078
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4228..4248)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4228..4246)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4267..4283)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4267..4283)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4280..4302)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4291..4307
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4315..4321,4322..4339,4340..4345))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4360..4381
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4498..4515)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(4669..4981,1..276))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4749..4768)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
       61 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
      121 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
      181 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
      241 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
      301 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
      361 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
      421 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
      481 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
      541 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacccttg
      601 gaggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
      661 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
      721 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
      781 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
      841 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
      901 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
      961 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     1021 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     1081 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     1141 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     1201 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     1261 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     1321 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     1381 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa
     1441 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     1501 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     1561 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     1621 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     1681 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     1741 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     1801 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     1861 ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact
     1921 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagtcgacc gccaattcaa
     1981 tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt gccaattcaa
     2041 tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca gccaattcaa
     2101 tatggcggac ttggcaccat gccaatatgg ctattggcca ggttcaatac tatgtattgg
     2161 ccctatgcca tatagtattc catatatggg ttttcctatt gacgtagata gcccctccca
     2221 atgggcggtc ccatatacca tatatggggc ttcctaatac cgcccatagc cactccccca
     2281 ttgacgtcaa tggtctctat atatggtctt tcctattgac gtcatatggg cggtcctatt
     2341 gacgtatatg gcgcctcccc cattgacgtc aattacggta aatggcccgc ctggctcaat
     2401 gcccattgac gtcaatagga ccacccacca ttgacgtcaa tgggatggct cattgcccat
     2461 tcatatccgt tctcacgccc cctattgacg tcaatgacgg taaatggccc acttggcagt
     2521 acatcaatat ctattaatag taacttggca agtacattac tattggaagt acgccagggt
     2581 acattggcag tactcccatt gacgtcaatg gcggtaaatg gcccgcgatg gctgccaagt
     2641 acatccccat tgacgtcaat ggggaggggc aatgacgcaa atgggcgttc cattgacgta
     2701 aatgggcggt aggcgtgcct aatgggaggt ctatataagc aatgctcgtt tagggaaccg
     2761 ccattctgcc tggggacgtc ggccttcgat caactcgcgc cgctggtcca ttctgcctgg
     2821 ggacgtcgga gcaagcttga tttaggtgac actatagaat acaagctact tgttcttttt
     2881 gcaggatcca ccatgatcaa gatcgccacc cggaaatacc tcggcaaaca gaacgtgtat
     2941 gatatcgggg tggagaggga tcataacttc gccctcaaaa atgggtttat tgcctctaat
     3001 tgtgccgacg gtttctacaa agatcggtat gtctatcggc atttcgcatc tgccgcactg
     3061 cctatcccgg aggtgctgga cataggtgaa tttagtgaat cactgactta ttgcatctcc
     3121 cgcagaagcc aaggcgtgac tcttcaggac cttccagaga ctgaactccc cgccgtcctg
     3181 cagcctgttg ctgaggccat ggacgccata gccgccgccg acctgagcca gacctctggt
     3241 tttgggcctt tcggccctca gggaattggc cagtacacca cctggaggga ctttatttgc
     3301 gccatcgccg acccccatgt gtatcactgg cagaccgtta tggacgatac cgtttccgca
     3361 agcgtggccc aagctctgga tgagctgatg ctgtgggcgg aggattgtcc cgaagtcagg
     3421 catcttgtcc acgccgactt tggatctaac aatgtcctga ctgataacgg cagaatcact
     3481 gccgtaatcg attggtcaga ggccatgttc ggcgactctc agtacgaggt ggctaacatc
     3541 ttcttttgga gaccgtggtt ggcctgtatg gaacagcaaa caagatactt cgagcgccgg
     3601 cacccggaac tggccggatc tccgaggctg agggcctaca tgctgaggat aggactggac
     3661 cagctctatc agtccctggt ggatggaaat ttcgacgatg cggcttgggc ccagggccgg
     3721 tgcgacgcaa tagtgcggag cggagctggg acagtgggcc ggacccagat tgcccggcga
     3781 tctgcagctg tgtggactga tggctgtgtt gaggtgctgg ctgacagcgg caatcggagg
     3841 cccagtacgc gacctagagc aaaaaaatac ccatacgatg ttccagatta cgcttatcct
     3901 tatgacgtac ctgactatgc atacccttat gatgtaccag actacgctta gtctagaact
     3961 atagtgagtc gtattacgta gatccagaca tgataagata cattgatgag tttggacaaa
     4021 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     4081 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     4141 tgtttcaggt tcagggggag gtgtgggagg ttttttaatt cgcggccgcg gcgccaatgc
     4201 attgggcccg gtacccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
     4261 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     4321 acgagccggg agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4381 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4441 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4501 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4561 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4621 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4681 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4741 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4801 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4861 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4921 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4981 g
//
