LOCUS       sequence-350058-        7214 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 7214)
  AUTHORS   Zhang Y, Cheng Y, Fang H, Roberts N, Zhang L, Vakulskas CA, Niedz 
            RP, Culver JN, Qi Y
  TITLE     Highly Efficient Genome Editing in Plant Protoplasts by 
            Ribonucleoprotein Delivery of CRISPR-Cas12a Nucleases.
  JOURNAL   Front Genome Ed. 2022 Jan 31;4:780238. doi: 
            10.3389/fgeed.2022.780238. eCollection 2022.
  PUBMED    35174354
REFERENCE   2  (bases 1 to 7214)
  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: "2022-01-31"; volume: "4:780238. doi"; pages: " 
            10.3389/fgeed.2022.780238. eCollection 2022"
COMMENT     Sequence Label: pYPQ220-Ultra
FEATURES             Location/Qualifiers
     source          1..7214
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             358..1149
                     /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      1242..1830
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1731..1750
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1984..2001
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2160..2187)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2279..2365)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2414..2436
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2428..2445
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2429..2445
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2461..2560
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2585..2605
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6548..6595
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      6664..6916
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    6961..7084
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(7101..7120)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7102..7120)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7125..7141)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7125..7141)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 tggaccagcc aggacagaaa tgcctcgact tcgctgctac ccaaggttgc cgggtgacgc
       61 acaccgtgga aacggatgaa ggcacgaacc cagtggacat aagcctgttc ggttcgtaag
      121 ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac cgaacgcagc
      181 ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt tggggtacag
      241 tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta
      301 tggagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt aaacattatg
      361 agggaagcgg tgatcgccga agtatcgact caactatcag aggtagttgg cgtcatcgag
      421 cgccatctcg aaccgacgtt gctggccgta catttgtacg gctccgcagt ggatggcggc
      481 ctgaagccac acagtgatat tgatttgctg gttacggtga ccgtaaggct tgatgaaaca
      541 acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg agagagcgag
      601 attctccgcg ctgtagaagt caccattgtt gtgcacgacg acatcattcc gtggcgttat
      661 ccagctaagc gcgaactgca atttggagaa tggcagcgca atgacattct tgcaggtatc
      721 ttcgagccag ccacgatcga cattgatctg gctatcttgc tgacaaaagc aagagaacat
      781 agcgttgcct tggtaggtcc agcggcggag gaactctttg atccggttcc tgaacaggat
      841 ctatttgagg cgctaaatga aaccttaacg ctatggaact cgccgcccga ctgggctggc
      901 gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa
      961 atcgcgccga aggatgtcgc tgccgactgg gcaatggagc gcctgccggc ccagtatcag
     1021 cccgtcatac ttgaagctag acaggcttat cttggacaag aagaagatcg cttggcctcg
     1081 cgcgcagatc agttggaaga atttgtccac tacgtgaaag gcgagatcac caaggtagtc
     1141 ggcaaataac cctcgagcca cccatgacca aaatccctta acgtgagtta cgcgtcgttc
     1201 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     1261 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     1321 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     1381 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     1441 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     1501 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     1561 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     1621 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     1681 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     1741 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     1801 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     1861 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     1921 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     1981 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     2041 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     2101 agtgagcgca acgcaattaa tacgcgtacc gcgagccagg aagagtttgt agaaacgcaa
     2161 aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc
     2221 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat
     2281 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
     2341 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgctt
     2401 gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc
     2461 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
     2521 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccgaattcgc ccttcaccat
     2581 ggctcctaag aagaagcgga aggttggtat tcacggggtg cctgcggcta cgcagttcga
     2641 ggggttcacc aacctctacc aggtcagcaa gacgctgcgg ttcgagctga ttccgcaggg
     2701 caagaccctc aagcacatcc aggagcaggg ctttatcgag gaggacaaag cgcggaacga
     2761 ccactacaag gagctcaagc cgatcatcga ccggatctac aagacctacg cggaccagtg
     2821 cctgcagctc gtgcagctcg actgggagaa cctctcggcc gccatcgact cctaccgcaa
     2881 ggagaagacg gaggagacac gcaacgcgct catcgaagaa caggctacgt atcgcaacgc
     2941 tatccacgac tacttcatag ggcgcacaga taatctcacc gacgccatca acaagcgcca
     3001 cgccgaaatc tataaggggc tgtttaaggc cgagctgttc aatggcaaag tgctgaagca
     3061 actgggcacc gtcacgacca cagagcatga gaacgccctc ctccggtcgt tcgacaagtt
     3121 tacgacatac tttagcggct tttacgagaa ccgcaaaaac gtgttcagcg ccgaggacat
     3181 tagcaccgcc atcccgcata ggatcgtgca agacaacttc ccgaagttca aggagaactg
     3241 ccacatcttc acccgcctca tcacggccgt gccgtccctc cgcgagcact tcgagaacgt
     3301 gaagaaggcc atcgggatct tcgtgtccac ctcgatcgaa gaggtgttct ccttcccctt
     3361 ctacaatcag ctgctgacgc agacccagat cgacctctac aaccagctcc tcggcggcat
     3421 ctcccgcgag gccggcaccg agaagatcaa gggcctgaac gaggtgctca atctcgccat
     3481 ccagaagaac gacgaaaccg cgcacatcat tgcctcactc ccacataggt ttatccccct
     3541 gtttaagcag atcctctccg accgcaacac gttgtccttc atcctcgagg agttcaagag
     3601 cgacgaggag gtcatccagt ccttctgcaa gtacaagacc ctcctccgca acgagaatgt
     3661 gctcgaaacc gccgaggcgc tgttcaatga gctcaatagc atcgatctca cccacatctt
     3721 catctcccat aagaagctgg aaaccatctc cagcgccctg tgcgaccact gggacaccct
     3781 ccgcaacgcc ctctacgagc ggcgcatcag cgagctcacc ggcaagatca cgaagtcggc
     3841 gaaagagaaa gtgcaaagga gcctcaagca cgaggacatt aacctgcagg agatcatctc
     3901 cgccgcgggc aaggagctgt ccgaggcttt caagcagaag acctcggaga tcctctctca
     3961 cgcccacgcg gccctggacc agccgctgcc gacgaccctg aaaaagcaag aggaaaagga
     4021 aatcctcaag tcccagctgg acagcctcct ggggctgtac caccttctcg actggttcgc
     4081 cgtggacgag tccaacgagg tcgatccgga gtttagcgcc cgcctcaccg ggatcaagct
     4141 tgagatggaa cctagcttga gcttctataa taaggcgcgc aattacgcga ccaagaagcc
     4201 gtattccgtg gagaagttca agctgaactt ccaacgcccc accctggcca gcgggtggga
     4261 cgttaacaaa gagaagaaca acggagccat tcttttcgtg aaaaatgggt tgtattattt
     4321 gggaattatg ccgaaacaaa aaggcaggta caaggcgctc agcttcgagc caactgagaa
     4381 gacctccgag ggcttcgaca agatgtacta cgattatttt cctgacgctg caaagatgat
     4441 accgaagtgc agcactcagc ttaaggcggt gacggcgcac tttcagaccc ataccacccc
     4501 catcctcctc tccaacaact tcatcgagcc gctcgagatc accaaggaga tatacgatct
     4561 gaataatcca gaaaaggaac ccaagaagtt ccagaccgcc tacgccaaga agacgggcga
     4621 tcaaaagggg tatagagagg cgctctgcaa gtggatcgac ttcacgcgcg atttcctcag
     4681 caagtacacc aagacaacct ccatcgatct ctcttccctc cgcccctctt cccagtacaa
     4741 ggacctcggg gagtactacg ccgaactcaa cccactcctg tatcacatct cgtttcagcg
     4801 tatcgcggaa aaggagatca tggacgccgt cgaaaccggc aagttgtatc ttttccaaat
     4861 ctataacaag gacttcgcga agggccacca cgggaagcca aacctgcaca ccctctactg
     4921 gacaggcctc ttcagcccgg aaaatctcgc gaagacgagc ataaagctga acggccaggc
     4981 agaactcttc tacaggccga agtccaggat gaagcgcatg gctcatcgcc tcggtgagaa
     5041 gatgctgaac aagaagctga aagatcaaaa gacgccaatc cctgatacac tgtatcagga
     5101 gctgtacgat tacgtgaacc accgcctctc acacgacctc agcgacgagg cccgcgcgct
     5161 cctgccaaac gtcatcacga aggaggtcag ccacgagatc ataaaggatc ggcggtttac
     5221 ctctgacaag ttcctcttcc atgtccccat cacgctgaac taccaggccg cgaatagccc
     5281 gtccaagttc aaccagcggg tcaacgcgta tctcaaggag cacccagaga cacccataat
     5341 cgggattgac cggggggagc ggaacctcat ctacatcacc gtcatcgact ccaccggaaa
     5401 gattctcgag caacggtcgc tcaataccat ccagcagttc gactaccaga agaagctcga
     5461 caaccgggag aaggaacgcg tcgccgcgag gcaggcctgg tccgtagtgg gcacgatcaa
     5521 agacctgaag cagggctatc tcagccaggt catccatgag atagtggatc tcatgatcca
     5581 ctaccaagcc gtcgtggtcc tcgagaatct caatttcgga ttcaaatcca agcgcacagg
     5641 catcgccgag aaggcggtgt accaacagtt cgagaaaatg cttatcgaca agctcaattg
     5701 cctggtgctc aaggactatc cggcggagaa ggtcgggggg gtcctcaatc cgtatcagct
     5761 gaccgaccag tttacgtcat ttgcgaagat gggcacccag agcggcttcc ttttctatgt
     5821 cccggcccca tatacctcaa agattgatcc cttgaccgga ttcgtggacc cgtttgtctg
     5881 gaagaccatc aagaaccatg agtcgcgtaa gcatttcctg gagggtttcg acttcctgca
     5941 ctatgatgta aaaaccggag acttcatcct gcatttcaag atgaatcgga acctctcctt
     6001 ccagcgggga ctccctggct tcatgcccgc ttgggatatc gtttttgaga aaaatgaaac
     6061 ccaattcgac gccaaaggca cgcctttcat cgcgggcaag aggattgtcc ctgtaattga
     6121 gaaccataga ttcaccgggc gttaccgtga cctgtacccc gcaaacgaac tcatcgccct
     6181 cctggaggag aaaggcatcg ttttccgcga cgggtcaaat atcctcccca aactgctcga
     6241 gaacgatgat agccacgcta ttgacacgat ggtagcgctc atcagatccg tgctgcaaat
     6301 gagaaattca aatgctgcca ctggggagga ttacatcaac tcccctgtgc gtgatctcaa
     6361 tggcgtgtgc ttcgattcta gatttcagaa tcctgagtgg ccgatggatg ccgatgctaa
     6421 cggcgcatac cacatagcat tgaaaggaca actgttgttg aaccatctca aggagagcaa
     6481 ggaccttaag ctgcagaacg gcatcagcaa ccaggattgg cttgcctata tccaagagct
     6541 ccgcaataag cgtcctgctg ccaccaaaaa ggccggacag gctaagaaaa agaagtgaga
     6601 cgactagtgg cggccgccgg atccacctga tctagagcta gcgtttaaac accggtgacg
     6661 tccgatcgtt caaacatttg gcaataaagt ttcttaagat tgaatcctgt tgccggtctt
     6721 gcgatgatta tcatataatt tctgttgaat tacgttaagc atgtaataat taacatgtaa
     6781 tgcatgacgt tatttatgag atgggttttt atgattagag tcccgcaatt atacatttaa
     6841 tacgcgatag aaaacaaaat atagcgcgca aactaggata aattatcgcg cgcggtgtca
     6901 tctatgttac tagatcggga attgatcccc cctcgacagc ttccggaaag ggcgaattcg
     6961 caactttgta tacaaaagtt gaacgagaaa cgtaaaatga tataaatatc aatatattaa
     7021 attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat atccagtcac
     7081 tatgccatcc agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct
     7141 ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat
     7201 catcatgcct cctc
//
