LOCUS       sequence-405375-        4430 bp    DNA     circular SYN 15-APR-2025
DEFINITION  p3flag_HygR(1-52)-NpuDnaE(N).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4430)
  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 4430)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pAXY0018
FEATURES             Location/Qualifiers
     source          1..4430
                     /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             91..114
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             118..141
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             145..168
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    672..806
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(696..715)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     750..769
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(919..939)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(919..937)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(958..974)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(958..974)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(971..993)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    982..998
                     /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(1006..1012,1013..1030,1031..1036))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1051..1072
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1189..1206)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1360..1948)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1440..1459)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2119..2910,2911..2979))
                     /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     2742..2761
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2980..3084)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3110..3565)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3247..3268)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3459..3478
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccaccatg gattataagg atgacgatga caaagcagac
      121 tacaaagacg acgatgataa ggctgattat aaagatgatg acgacaaagg ccggccaatg
      181 aagaaaccgg aactgactgc aacatctgtc gagaagtttc tgattgaaaa gttcgattct
      241 gtttccgatc tcatgcagct cagtgagggc gaagagtcca gagctttctc attcgacgtg
      301 ggcgggcgcg gatacgtact gcgagttaat tcttgcctga gttatgagac tgagattctg
      361 accgtagaat atggtctgct tccaatcggc aagattgtgg aaaaaaggat cgagtgtacc
      421 gtgtattctg tggacaataa cggcaatatt tacacccaac ctgtggcaca gtggcatgac
      481 cggggggagc aggaagtttt tgaatactgc ttggaggatg gatccctcat tagagctaca
      541 aaagatcaca agtttatgac cgtagacgga cagatgttgc caatcgacga gatctttgag
      601 agggagctgg atctgatgcg ggtggacaat ctgcccaact agtctagaac tatagtgagt
      661 cgtattacgt agatccagac atgataagat acattgatga gtttggacaa accacaacta
      721 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
      781 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
      841 ttcaggggga ggtgtgggag gttttttaat tcgcggccgc ggcgccaatg cattgggccc
      901 ggtacccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
      961 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     1021 gagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     1081 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     1141 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     1201 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     1261 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     1321 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1381 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1441 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1501 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1561 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1621 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1681 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1741 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1801 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1861 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1921 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     1981 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2041 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2101 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2161 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     2221 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccaccct tggaggctcc
     2281 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     2341 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     2401 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     2461 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     2521 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     2581 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     2641 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     2701 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     2761 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     2821 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     2881 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     2941 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     3001 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     3061 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc
     3121 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     3181 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     3241 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     3301 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     3361 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     3421 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     3481 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     3541 cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa
     3601 gggcgatcgg tgcgggcctc ttcgctatta cgccagtcga ccgccaattc aatatggcgt
     3661 atatggactc atgccaattc aatatggtgg atctggacct gtgccaattc aatatggcgt
     3721 atatggactc gtgccaattc aatatggtgg atctggaccc cagccaattc aatatggcgg
     3781 acttggcacc atgccaatat ggctattggc caggttcaat actatgtatt ggccctatgc
     3841 catatagtat tccatatatg ggttttccta ttgacgtaga tagcccctcc caatgggcgg
     3901 tcccatatac catatatggg gcttcctaat accgcccata gccactcccc cattgacgtc
     3961 aatggtctct atatatggtc tttcctattg acgtcatatg ggcggtccta ttgacgtata
     4021 tggcgcctcc cccattgacg tcaattacgg taaatggccc gcctggctca atgcccattg
     4081 acgtcaatag gaccacccac cattgacgtc aatgggatgg ctcattgccc attcatatcc
     4141 gttctcacgc cccctattga cgtcaatgac ggtaaatggc ccacttggca gtacatcaat
     4201 atctattaat agtaacttgg caagtacatt actattggaa gtacgccagg gtacattggc
     4261 agtactccca ttgacgtcaa tggcggtaaa tggcccgcga tggctgccaa gtacatcccc
     4321 attgacgtca atggggaggg gcaatgacgc aaatgggcgt tccattgacg taaatgggcg
     4381 gtaggcgtgc ctaatgggag gtctatataa gcaatgctcg tttagggaac
//
