LOCUS       sequence-405374-        4823 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pNpuDnaE(C)_HygR(90-341)_3HA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4823)
  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 4823)
  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: pAXY0017
FEATURES             Location/Qualifiers
     source          1..4823
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             952..1032
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     polyA_signal    1065..1199
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1089..1108)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1143..1162
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1312..1332)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1312..1330)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1351..1367)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1351..1367)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1364..1386)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1375..1391
                     /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(1399..1405,1406..1423,1424..1429))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1444..1465
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1582..1599)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1753..2341)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1833..1852)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2512..3303,3304..3372))
                     /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     3135..3154
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3373..3477)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3503..3958)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3640..3661)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3852..3871
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccaccatg atcaagatcg ccacccggaa atacctcggc
      121 aaacagaacg tgtatgatat cggggtggag agggatcata acttcgccct caaaaatggg
      181 tttattgcct ctaattgcat ctcccgcaga agccaaggcg tgactcttca ggaccttcca
      241 gagactgaac tccccgccgt cctgcagcct gttgctgagg ccatggacgc catagccgcc
      301 gccgacctga gccagacctc tggttttggg cctttcggcc ctcagggaat tggccagtac
      361 accacctgga gggactttat ttgcgccatc gccgaccccc atgtgtatca ctggcagacc
      421 gttatggacg ataccgtttc cgcaagcgtg gcccaagctc tggatgagct gatgctgtgg
      481 gcggaggatt gtcccgaagt caggcatctt gtccacgccg actttggatc taacaatgtc
      541 ctgactgata acggcagaat cactgccgta atcgattggt cagaggccat gttcggcgac
      601 tctcagtacg aggtggctaa catcttcttt tggagaccgt ggttggcctg tatggaacag
      661 caaacaagat acttcgagcg ccggcacccg gaactggccg gatctccgag gctgagggcc
      721 tacatgctga ggataggact ggaccagctc tatcagtccc tggtggatgg aaatttcgac
      781 gatgcggctt gggcccaggg ccggtgcgac gcaatagtgc ggagcggagc tgggacagtg
      841 ggccggaccc agattgcccg gcgatctgca gctgtgtgga ctgatggctg tgttgaggtg
      901 ctggctgaca gcggcaatcg gaggcccagt acgcgaccta gagcaaaaaa atacccatac
      961 gatgttccag attacgctta tccttatgac gtacctgact atgcataccc ttatgatgta
     1021 ccagactacg cttagtctag aactatagtg agtcgtatta cgtagatcca gacatgataa
     1081 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     1141 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     1201 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     1261 aattcgcggc cgcggcgcca atgcattggg cccggtaccc agcttttgtt ccctttagtg
     1321 agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     1381 tccgctcaca attccacaca acatacgagc cgggagcata aagtgtaaag cctggggtgc
     1441 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     1501 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     1561 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     1621 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     1681 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     1741 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1801 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1861 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1921 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     1981 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2041 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2101 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2161 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     2221 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2281 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2341 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     2401 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     2461 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     2521 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     2581 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     2641 aatgataccg cgagacccac ccttggaggc tccagattta tcagcaataa accagccagc
     2701 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     2761 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     2821 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     2881 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     2941 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3001 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     3061 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     3121 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     3181 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     3241 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     3301 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     3361 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     3421 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     3481 atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta
     3541 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     3601 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     3661 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     3721 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     3781 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     3841 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     3901 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
     3961 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     4021 ttacgccagt cgaccgccaa ttcaatatgg cgtatatgga ctcatgccaa ttcaatatgg
     4081 tggatctgga cctgtgccaa ttcaatatgg cgtatatgga ctcgtgccaa ttcaatatgg
     4141 tggatctgga ccccagccaa ttcaatatgg cggacttggc accatgccaa tatggctatt
     4201 ggccaggttc aatactatgt attggcccta tgccatatag tattccatat atgggttttc
     4261 ctattgacgt agatagcccc tcccaatggg cggtcccata taccatatat ggggcttcct
     4321 aataccgccc atagccactc ccccattgac gtcaatggtc tctatatatg gtctttccta
     4381 ttgacgtcat atgggcggtc ctattgacgt atatggcgcc tcccccattg acgtcaatta
     4441 cggtaaatgg cccgcctggc tcaatgccca ttgacgtcaa taggaccacc caccattgac
     4501 gtcaatggga tggctcattg cccattcata tccgttctca cgccccctat tgacgtcaat
     4561 gacggtaaat ggcccacttg gcagtacatc aatatctatt aatagtaact tggcaagtac
     4621 attactattg gaagtacgcc agggtacatt ggcagtactc ccattgacgt caatggcggt
     4681 aaatggcccg cgatggctgc caagtacatc cccattgacg tcaatgggga ggggcaatga
     4741 cgcaaatggg cgttccattg acgtaaatgg gcggtaggcg tgcctaatgg gaggtctata
     4801 taagcaatgc tcgtttaggg aac
//
