LOCUS       sequence-414100-        7149 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry plasmid (attL1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7149)
  AUTHORS   Li G, Zhang Y, Dailey M, Qi Y
  TITLE     Hs1Cas12a and Ev1Cas12a confer efficient genome editing in plants.
  JOURNAL   Front Genome Ed. 2023 Oct 12;5:1251903. doi: 
            10.3389/fgeed.2023.1251903. eCollection 2023.
  PUBMED    37901282
REFERENCE   2  (bases 1 to 7149)
  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: "Front 
            Genome Ed."; date: "2023-10-12"; volume: "5:1251903. doi"; pages: " 
            10.3389/fgeed.2023.1251903. eCollection 2023"
COMMENT     Sequence Label: pYPQ301
FEATURES             Location/Qualifiers
     source          1..7149
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             378..1169
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      1262..1850
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1751..1770
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2004..2021
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2180..2207)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2299..2385)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2434..2456
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2448..2465
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2449..2465
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2481..2580
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2605..2625
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2931..2942
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             6541..6588
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      6619..6871
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    6916..7039
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(7056..7075)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7057..7075)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7080..7096)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7080..7096)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atatatcatc atgcctcctc tggaccagcc aggacagaaa tgcctcgact tcgctgctac
       61 ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc cagtggacat
      121 aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc aactggtcca
      181 gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg
      241 actgtttttt tggggtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg
      301 tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagggc agtcgcccta
      361 aaacaaagtt aaacattatg agggaagcgg tgatcgccga agtatcgact caactatcag
      421 aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt gctggccgta catttgtacg
      481 gctccgcagt ggatggcggc ctgaagccac acagtgatat tgatttgctg gttacggtga
      541 ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg
      601 cttcccctgg agagagcgag attctccgcg ctgtagaagt caccattgtt gtgcacgacg
      661 acatcattcc gtggcgttat ccagctaagc gcgaactgca atttggagaa tggcagcgca
      721 atgacattct tgcaggtatc ttcgagccag ccacgatcga cattgatctg gctatcttgc
      781 tgacaaaagc aagagaacat agcgttgcct tggtaggtcc agcggcggag gaactctttg
      841 atccggttcc tgaacaggat ctatttgagg cgctaaatga aaccttaacg ctatggaact
      901 cgccgcccga ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt
      961 acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg gcaatggagc
     1021 gcctgccggc ccagtatcag cccgtcatac ttgaagctag acaggcttat cttggacaag
     1081 aagaagatcg cttggcctcg cgcgcagatc agttggaaga atttgtccac tacgtgaaag
     1141 gcgagatcac caaggtagtc ggcaaataac cctcgagcca cccatgacca aaatccctta
     1201 acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     1261 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     1321 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     1381 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     1441 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     1501 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     1561 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     1621 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     1681 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     1741 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     1801 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     1861 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     1921 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     1981 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     2041 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     2101 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc gcgagccagg
     2161 aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct tagtttgatg
     2221 cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt
     2281 caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat
     2341 aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc tggcagttcc
     2401 ctactctcgc gttaacgctt gcatggatgt tttcccagtc acgacgttgt aaaacgacgg
     2461 ccagtcttaa gctcgggccc caaataatga ttttattttg actgatagtg acctgttcgt
     2521 tgcaacaaat tgatgagcaa tgctttttta taatgccaac tttgtacaaa aaagcaggct
     2581 ccgaattcgc ccttcaccat ggctcctaag aagaagcgga aggttggtat tcacggggtg
     2641 cctgcggcta ccaagacatt cgactctgaa tttttcaacc tttacagtct tcagaaaacc
     2701 gtcagatttg agctcaaacc ggtcggtgaa acggcctcat tcgtcgagga ctttaagaac
     2761 gaaggcctga agcgggttgt tagcgaagac gagaggcgcg cagtggatta tcaaaaagtg
     2821 aaggaaatca ttgacgatta tcatcgcgat ttcattgaag agtcactgaa ctacttccca
     2881 gaacaggttt caaaagacgc cttggaacag gcttttcact tgtaccaaaa attgaaggcc
     2941 gccaaggtgg aagaacggga gaaagccttg aaggagtggg aagcgctcca aaaaaaattg
     3001 agggagaaag tggtgaaatg ctttagtgat tcaaacaagg ccaggtttag tagaatagat
     3061 aaaaaggaac tcataaagga ggatcttatt aattggctcg tggctcaaaa ccgcgaagac
     3121 gacattccca cagtggaaac cttcaacaac ttcaccactt atttcaccgg gtttcacgaa
     3181 aatagaaaga atatatacag caaggatgac catgcaaccg ctatatcttt ccgcttgata
     3241 cacgaaaacc tcccgaaatt cttcgacaat gtcatctcgt tcaacaagct caaggaaggt
     3301 tttcccgaat tgaaattcga taaggtgaaa gaagacttgg aggtggacta tgacctgaaa
     3361 catgcctttg aaatagaata ttttgtcaat ttcgtgacac aagcggggat cgatcaatat
     3421 aattacttgc tgggcggcaa gacgttggag gatggcacta aaaagcaggg catgaatgag
     3481 cagatcaatc ttttcaagca acagcaaact agagacaagg ctagacagat cccaaaattg
     3541 atcccattgt tcaaacagat cctttcggag aggaccgaat ctcagtcgtt tatccccaaa
     3601 cagttcgaaa gtgaccagga actgtttgat tcccttcaaa aattgcataa caactgtcag
     3661 gacaagttca ctgtcttgca gcaagccatc ttgggcctgg cggaagcaga tttgaaaaaa
     3721 gtgtttataa agacctcgga cttgaatgcg cttagcaata cgatttttgg caattactct
     3781 gtgtttagcg atgcactcaa cctttataaa gagagcctga agaccaagaa agcccaagaa
     3841 gcttttgaga aacttcctgc gcattcgatc catgatctta ttcaatatct tgagcagttt
     3901 aactcaagtc ttgacgcgga aaagcagcag tccactgaca cggttctgaa ctatttcatc
     3961 aaaacagatg aactctactc gagatttatt aaatcaacct cggaggcatt tactcaggtg
     4021 caaccgcttt tcgagctcga agccctttcc agcaaacgca gaccaccgga atccgaagat
     4081 gaaggtgcta aaggacaaga agggttcgaa caaattaagc ggattaaggc atatcttgat
     4141 acgcttatgg aagcggtcca cttcgcgaag ccgctttacc tggtcaaggg gcggaaaatg
     4201 atcgagggtc ttgacaaaga tcagtctttt tacgaggcgt ttgagatggc ttaccaggag
     4261 ctcgaaagtt tgataatccc aatatacaac aaggccagat cctatctctc gcggaagcct
     4321 tttaaggcag ataagttcaa gattaacttt gacaacaaca cgcttttgtc aggctgggac
     4381 gccaataagg aaactgccaa cgcgagtatc ctctttaaaa aagacggtct ttactatctt
     4441 ggtataatgc caaaaggtaa aacatttttg tttgattatt tcgtcagctc ggaagattcc
     4501 gaaaaactta aacagcggag gcaaaagacg gccgaggagg ccctcgcgca ggatggtgag
     4561 agctattttg aaaaaattcg ctataagttg ctccccggag ctagtaaaat gttgcccaag
     4621 gtcttcttta gtaataagaa cattggcttc tacaatccct cagatgatat ccttaggata
     4681 aggaatacag cgagccacac gaaaaacggc acaccgcaaa aaggacattc caaagtggag
     4741 ttcaatctga acgattgcca taagatgatc gatttcttta aatcctctat ccaaaaacac
     4801 cccgaatggg gctcgttcgg tttcactttt tcggacacga gtgattttga agacatgagt
     4861 gcgttctata gggaggttga aaatcaaggg tatgtcatct cgtttgataa gattaaagaa
     4921 acgtatattc aaagccaggt tgaacagggc aatctctatc ttttccagat atacaataaa
     4981 gacttttcgc catactctaa gggaaagccg aatctccata ctctgtattg gaaagccctc
     5041 ttcgaggaag cgaaccttaa caatgttgtg gcaaagctta atggggaagc agaaatattt
     5101 ttccgccgcc actcaataaa ggccagcgac aaagttgtgc atcctgcgaa ccaggccatt
     5161 gataataaga acccgcacac agaaaagact cagagtacct tcgaatatga tcttgttaaa
     5221 gacaagcgct acacacagga caagttcttt ttccacgttc caatatcact caatttcaag
     5281 gcgcaaggag tcagcaaatt taatgataaa gtcaatggat tcctcaaagg gaatccggat
     5341 gtgaacatta ttgggataga cagaggcgag agacacctgc tgtactttac ggttgtgaat
     5401 caaaaggggg aaattttggt ccaggagtct ctcaacacgc tcatgagtga caagggccac
     5461 gtcaatgatt accaacagaa gctggacaaa aaagaacagg agagggatgc agcaagaaaa
     5521 agctggacga cggtggagaa tatcaaggag ctgaaagaag gttacttgtc gcacgttgtg
     5581 cataagctgg ctcatctgat cattaaatac aatgccatcg tctgcttgga ggacctcaac
     5641 tttggtttta aacgcgggag atttaaggtc gagaaacagg tttaccaaaa gttcgaaaaa
     5701 gcgctcatcg ataagctgaa ctacctggtg tttaaagaaa aagaacttgg ggaagtcggg
     5761 cactatctca cagcgtacca gcttacagcg ccttttgaga gcttcaaaaa actcggcaaa
     5821 cagtcgggca tccttttcta cgtcccggcg gattatacgt caaagataga ccccacaaca
     5881 gggtttgtta acttcttgga tcttcggtat caaagcgtcg aaaaggctaa gcaacttctc
     5941 tccgacttca acgctataag gtttaatagc gtccaaaatt atttcgagtt tgaaatagat
     6001 tataagaagc tcacaccaaa gcggaaggtt gggacccagt ctaagtgggt catatgcaca
     6061 tacggcgatg tgaggtacca gaataggcgc aatcaaaagg gtcactggga gaccgaagag
     6121 gttaatgtga ctgagaagct taaggctctt tttgcctcag actctaaaac aactactgtg
     6181 atcgattacg ccaacgatga caacctcatt gacgtgatcc tcgaacaaga taaggcctcc
     6241 ttttttaaag aactcctttg gcttctgaag ctgacgatga cactccgcca ctccaaaatc
     6301 aagagcgagg acgactttat actgtcacct gtgaagaatg aacaaggcga attttacgac
     6361 tcgaggaaag caggcgaagt ttggcccaaa gacgctgacg ccaatggggc atatcacatc
     6421 gctcttaagg gactctggaa cttgcaacag attaatcagt gggagaaagg gaagaccttg
     6481 aatctcgcga taaagaatca agattggttc agctttattc aagaaaagcc gtaccaagaa
     6541 aagcgtcctg ctgccaccaa aaaggccgga caggctaaga aaaagaagtg agacgactag
     6601 tggcggccgc cgacgtccga tcgttcaaac atttggcaat aaagtttctt aagattgaat
     6661 cctgttgccg gtcttgcgat gattatcata taatttctgt tgaattacgt taagcatgta
     6721 ataattaaca tgtaatgcat gacgttattt atgagatggg tttttatgat tagagtcccg
     6781 caattataca tttaatacgc gatagaaaac aaaatatagc gcgcaaacta ggataaatta
     6841 tcgcgcgcgg tgtcatctat gttactagat cgggaattga tcccccctcg acagcttccg
     6901 gaaagggcga attcgcaact ttgtatacaa aagttgaacg agaaacgtaa aatgatataa
     6961 atatcaatat attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac
     7021 aacatatcca gtcactatgc catccagctg atatccccta tagtgagtcg tattacatgg
     7081 tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac
     7141 aagataaaa
//
