LOCUS       sequence-405373-        4541 bp    DNA     circular SYN 14-APR-2025
DEFINITION  p3flag_HygR(1-89)-NpuDnaE(N).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4541)
  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 4541)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pAXY0016
FEATURES             Location/Qualifiers
     source          1..4541
                     /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    783..917
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(807..826)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     861..880
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1030..1050)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1030..1048)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1069..1085)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1069..1085)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1082..1104)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1093..1109
                     /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(1117..1123,1124..1141,1142..1147))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1162..1183
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1300..1317)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1471..2059)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1551..1570)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2230..3021,3022..3090))
                     /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     2853..2872
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3091..3195)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3221..3676)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3358..3379)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3570..3589
                     /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 tcttgtgccg acggtttcta caaagatcgg
      361 tatgtctatc ggcatttcgc atctgccgca ctgcctatcc cggaggtgct ggacataggt
      421 gaatttagtg aatcactgac ttattgcctg agttatgaga ctgagattct gaccgtagaa
      481 tatggtctgc ttccaatcgg caagattgtg gaaaaaagga tcgagtgtac cgtgtattct
      541 gtggacaata acggcaatat ttacacccaa cctgtggcac agtggcatga ccggggggag
      601 caggaagttt ttgaatactg cttggaggat ggatccctca ttagagctac aaaagatcac
      661 aagtttatga ccgtagacgg acagatgttg ccaatcgacg agatctttga gagggagctg
      721 gatctgatgc gggtggacaa tctgcccaac tagtctagaa ctatagtgag tcgtattacg
      781 tagatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
      841 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
      901 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
      961 aggtgtggga ggttttttaa ttcgcggccg cggcgccaat gcattgggcc cggtacccag
     1021 cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt
     1081 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg ggagcataaa
     1141 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     1201 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     1261 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     1321 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     1381 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     1441 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     1501 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1561 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     1621 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     1681 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     1741 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     1801 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     1861 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     1921 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     1981 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2041 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     2101 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     2161 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     2221 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     2281 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     2341 atctggcccc agtgctgcaa tgataccgcg agacccaccc ttggaggctc cagatttatc
     2401 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     2461 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     2521 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     2581 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     2641 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     2701 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     2761 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     2821 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     2881 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     2941 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     3001 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     3061 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     3121 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     3181 aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt
     3241 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     3301 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     3361 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     3421 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     3481 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     3541 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     3601 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     3661 ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg
     3721 gtgcgggcct cttcgctatt acgccagtcg accgccaatt caatatggcg tatatggact
     3781 catgccaatt caatatggtg gatctggacc tgtgccaatt caatatggcg tatatggact
     3841 cgtgccaatt caatatggtg gatctggacc ccagccaatt caatatggcg gacttggcac
     3901 catgccaata tggctattgg ccaggttcaa tactatgtat tggccctatg ccatatagta
     3961 ttccatatat gggttttcct attgacgtag atagcccctc ccaatgggcg gtcccatata
     4021 ccatatatgg ggcttcctaa taccgcccat agccactccc ccattgacgt caatggtctc
     4081 tatatatggt ctttcctatt gacgtcatat gggcggtcct attgacgtat atggcgcctc
     4141 ccccattgac gtcaattacg gtaaatggcc cgcctggctc aatgcccatt gacgtcaata
     4201 ggaccaccca ccattgacgt caatgggatg gctcattgcc cattcatatc cgttctcacg
     4261 ccccctattg acgtcaatga cggtaaatgg cccacttggc agtacatcaa tatctattaa
     4321 tagtaacttg gcaagtacat tactattgga agtacgccag ggtacattgg cagtactccc
     4381 attgacgtca atggcggtaa atggcccgcg atggctgcca agtacatccc cattgacgtc
     4441 aatggggagg ggcaatgacg caaatgggcg ttccattgac gtaaatgggc ggtaggcgtg
     4501 cctaatggga ggtctatata agcaatgctc gtttagggaa c
//
