LOCUS       sequence-368895-        9194 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cre recombinase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9194)
  AUTHORS   Aliu E, Lee K, Wang K
  TITLE     CRISPR RNA-guided integrase enables high-efficiency targeted genome 
            engineering in Agrobacterium tumefaciens.
  JOURNAL   Plant Biotechnol J. 2022 Jun 11. doi: 10.1111/pbi.13872.
  PUBMED    35690588
REFERENCE   2  (bases 1 to 9194)
  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: "Plant 
            Biotechnol J. 2022 Jun 11. doi: 10.1111/pbi.13872."
COMMENT     Sequence Label: pKL2315
FEATURES             Location/Qualifiers
     source          1..9194
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..25
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    103..277
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        324..358
                     /label=BBa_J23107
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.36"
     RBS             368..379
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      complement(1451..1482)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        complement(2937..3005)
                     /label=recA (Delta-LexA) promoter
                     /note="constitutively active E. coli recA promoter lacking 
                     the LexA binding site (Brent and Ptashne, 1981)"
     CDS             4240..4869
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             5298..6371
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      6437..6631
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(6867..6889)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6975..7115
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     7030..7049
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7130..7147)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7301..7889)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7381..7400)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7976..8770)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
        1 tggcaggata tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg
       61 gacgttttta atgtactgaa ttaacgccga attaattcgg gggatctgga ttttagtact
      121 ggattttggt tttaggaatt agaaatttta ttgatagaag tattttacaa atacaaatac
      181 atactaaggg tttcttatat gctcaacaca tgagcgaaac cctataggaa ccctaattcc
      241 cttatctggg aactactcac acattattat ggagaaactc gagtcaaatc tcggtgacgg
      301 gcaggaccgg acggggcggt acctttacgg ctagctcagc cctaggtatt atgctagcga
      361 attcattaaa gaggagaaaa cgcgtatgtc caacctgctc acggttcacc agaaccttcc
      421 ggctcttcca gtggacgcga cgtccgatga agtcaggaag aacctcatgg acatgttccg
      481 cgacaggcaa gcgttcagcg agcacacctg gaagatgctg ctctccgtct gccgctcctg
      541 ggctgcatgg tgcaagctga acaacaggaa gtggttcccc gctgagcccg aggacgtgag
      601 ggattacctt ctgtacctgc aagcgcgagg tctggcagtg aagaccatcc agcaacacct
      661 tggacaactg aacatgcttc acaggcgctc cggcctcccg cgccccagcg actcgaacgc
      721 cgtgagcctc gtcatgcgcc gcatcaggaa ggaaaacgtc gatgccggcg aaagggcaaa
      781 gcaggccctc gcgttcgaga gtaccgattt cgaccaggtc cgcagcctga tggagaacag
      841 cgacaggtgc caggacatta ggaacctggc gttcctcgga attgcataca acacgctcct
      901 caggatcgcg gaaattgccc gcattcgcgt gaaggacatt agccgcaccg acggcggcag
      961 gatgcttatc cacattggca ggaccaagac gctcgtttcc accgcaggcg tcgaaaaggc
     1021 cctcagcctc ggagtgacca agctcgtcga acgctggatc tccgtgtccg gcgtcgcgga
     1081 cgacccaaac aactacctct tctgccgcgt ccgcaagaac ggggtggctg cccctagcgc
     1141 caccagccaa ctcagcacga gggccttgga aggtattttc gaggccaccc accgcctgat
     1201 ctacggcgcg aaggatgaca gcggtcaacg ctacctcgca tggtccgggc actccgcccg
     1261 cgttggagct gctagggaca tggcccgcgc cggtgtttcc atccccgaaa tcatgcaggc
     1321 gggtggatgg acgaacgtga acattgtcat gaactacatt cgcaaccttg acagcgagac
     1381 gggcgcaatg gttcgcctcc tggaagatgg tgactgaaag cttgagaccg ttaattaaca
     1441 accaaaaaaa agtcaaaagc ctccggtcgg aggcttttga ctttctgctt attagttgac
     1501 tgtcagctgt ccttgctcca ggatgctgtt tttgacaacg gatgttttat tgcctttgat
     1561 gttcattaag aaagatggcg caaatgttgc ctttttatcc tcgaagaagc ctctgtttgt
     1621 catgtagctt gtgataacca cattgttgcc tttggcttgc ggcactgcga agtgagagta
     1681 agtgaatgtc acatcgtttg gatcaagacc catttgcagc acaagccctg ttttgttcag
     1741 cggcttgtaa gggccggtta aagagtttga tacataacca agcatgtaaa tatcgtttga
     1801 gttaatacca tcgatcgtca tttttgaacc gcgtgaatca gtgaacaagt accatttgcc
     1861 gttcattttg aaaacattcg cgcgctcgat ttcatcagtt accgtgtttg aagtgatcag
     1921 cggcttcatt acttttttca atgtgtaatc attatttaac tctatgatac cgagggcgcc
     1981 gttcgctaac tcagcatcgc gttttttagc gctctgctga agtttctggc tttctttacg
     2041 gaagaagttc gtgccgccgc cgtagtacgc tttgttaaat aaagattctt cgccttggta
     2101 tccgttttct gttcccgtgt tggcttcgaa tacaaggtat ttatggcctt tgtcttcaac
     2161 gtagtgaggg tcacgcagcg tatggttgtc gccggatgta taattgcctt catcaataaa
     2221 ctgctgaacg ttctgatatg tttttccgtc tccgtcaaaa atcgttttgt gatcttccac
     2281 tccgttgatt ttgagtgtgt catcagattt tgacacattt acctgcgctg ttgtcaggct
     2341 ttgtttgccg taatgtttac cggaatagtc agtgtagaat aaacggattt ttccgtcaga
     2401 tgtaaaggtt gcagaaccgg accattcttg cgtctgatct ttcaggatcg gatcgttggc
     2461 gtcgaactta tcgctgtctt taaagacacg gcccgcgttt ttccagctgt cgattgagtt
     2521 gtcgccgacc ttttgataaa acatgtagat tgatgtgtca tcagcgtctt tcgggcttcc
     2581 cgcaagagca aacacaacgt gatagccgtt gtattcagct acggttccgt cagcgttttg
     2641 cagcggccag ctgtcccaca catcaagtcc ttttgcagac tcaatatttt taatcgttga
     2701 ttgatcgaat tgaggcactt ggtatttttc gttttgctgc tgtttaggga tttgcagcat
     2761 atcatggcgt gtaatatgag agacgccata cgtttctttg tatgcttttt ggttattttc
     2821 tttcgcgaag gcttgagtcg ctcctcctgc cagaagtgca gtcgtaaaag tcagaactgt
     2881 ggcttgtttt acaatttttt tgatgttcat gttcatgtct cctcctgtat gtactggccg
     2941 ggtaataccg gatagtcaat atgttctgtt gaagcaatta tagcagctac gtggacctta
     3001 ttaagcgatc gagcacacgg gcatgccaac cacagggttc ccctcgggat caaagtactt
     3061 tgatccaacc cctccgctgc tatagtgcag tcggcttctg acgttcagtg cagccgtctt
     3121 ctgaaaacga catgtcgcac aagtcctaag ttacgcgaca ggctgccgcc ctgccctttt
     3181 cctggcgttt tcttgtcgcg tgttttagtc gcataaagta gaatacttgc gactagaacc
     3241 ggagacatta cgccatgaac aagagcgccg ccgctggcct gctgggctat gcccgcgtca
     3301 gcaccgacga ccaggacttg accaaccaac gggccgaact gcacgcggcc ggctgcacca
     3361 agctgttttc cgagaagatc accggcacca ggcgcgaccg cccggagctg gccaggatgc
     3421 ttgaccacct acgccctggc gacgttgtga cagtgaccag gctagaccgc ctggcccgca
     3481 gcacccgcga cctactggac attgccgagc gcatccagga ggccggcgcg ggcctgcgta
     3541 gcctggcaga gccgtgggcc gacaccacca cgccggccgg ccgcatggtg ttgaccgtgt
     3601 tcgccggcat tgccgagttc gagcgttccc taatcatcga ccgcacccgg agcgggcgcg
     3661 aggccgccaa ggcccgaggc gtgaagtttg gcccccgccc taccctcacc ccggcacaga
     3721 tcgcgcacgc ccgcgagctg atcgaccagg aaggccgcac cgtgaaagag gcggctgcac
     3781 tgcttggcgt gcatcgctcg accctgtacc gcgcacttga gcgcagcgag gaagtgacgc
     3841 ccaccgaggc caggcggcgc ggtgccttcc gtgaggacgc attgaccgag gccgacgccc
     3901 tggcggccgc cgagaatgaa cgccaagagg aacaagcatg aaaccgcacc aggacggcca
     3961 ggacgaaccg tttttcatta ccgaagagat cgaggcggag atgatcgcgg ccgggtacgt
     4021 gttcgagccg cccgcgcacg tctcaaccgt gcggctgcat gaaatcctgg ccggtttgtc
     4081 tgatgccaag ctggcggcct ggccggccag cttggccgct gaagaaaccg agcgccgccg
     4141 tctaaaaagg tgatgtgtat ttgagtaaaa cagcttgcgt catgcggtcg ctgcgtatat
     4201 gatgcgatga gtaaataaac aaatacgcaa ggggaacgca tgaaggttat cgctgtactt
     4261 aaccagaaag gcgggtcagg caagacgacc atcgcaaccc atctagcccg cgccctgcaa
     4321 ctcgccgggg ccgatgttct gttagtcgat tccgatcccc agggcagtgc ccgcgattgg
     4381 gcggccgtgc gggaagatca accgctaacc gttgtcggca tcgaccgccc gacgattgac
     4441 cgcgacgtga aggccatcgg ccggcgcgac ttcgtagtga tcgacggagc gccccaggcg
     4501 gcggacttgg ctgtgtccgc gatcaaggca gccgacttcg tgctgattcc ggtgcagcca
     4561 agcccttacg acatatgggc caccgccgac ctggtggagc tggttaagca gcgcattgag
     4621 gtcacggatg gaaggctaca agcggccttt gtcgtgtcgc gggcgatcaa aggcacgcgc
     4681 atcggcggtg aggttgccga ggcgctggcc gggtacgagc tgcccattct tgagtcccgt
     4741 atcacgcagc gcgtgagcta cccaggcact gccgccgccg gcacaaccgt tcttgaatca
     4801 gaacccgagg gcgacgctgc ccgcgaggtc caggcgctgg ccgctgaaat taaatcaaaa
     4861 ctcatttgag ttaatgaggt aaagagaaaa tgagcaaaag cacaaacacg ctaagtgccg
     4921 gccgtccgag cgcacgcagc agcaaggctg caacgttggc cagcctggca gacacgccag
     4981 ccatgaagcg ggtcaacttt cagttgccgg cggaggatca caccaagctg aagatgtacg
     5041 cggtacgcca aggcaagacc attaccgagc tgctatctga atacatcgcg cagctaccag
     5101 agtaaatgag caaatgaata aatgagtaga tgaattttag cggctaaagg aggcggcatg
     5161 gaaaatcaag aacaaccagg caccgacgcc gtggaatgcc ccatgtgtgg aggaacgggc
     5221 ggttggccag gcgtaagcgg ctgggttgcc tgccggccct gcaatggcac tggaaccccc
     5281 aagcccgagg aatcggcgtg agcggtcgca aaccatccgg cccggtacaa atcggcgcgg
     5341 cgctgggtga tgacctggtg gagaagttga aggccgcgca ggccgcccag cggcaacgca
     5401 tcgaggcaga agcacgcccc ggtgaatcgt ggcaagcggc cgctgatcga atccgcaaag
     5461 aatcccggca accgccggca gccggtgcgc cgtcgattag gaagccgccc aagggcgacg
     5521 agcaaccaga ttttttcgtt ccgatgctct atgacgtggg cacccgcgat agtcgcagca
     5581 tcatggacgt ggccgttttc cgtctgtcga agcgtgaccg acgagctggc gaggtgatcc
     5641 gctacgagct tccagacggg cacgtagagg tttccgcagg gccggccggc atggccagtg
     5701 tgtgggatta cgacctggta ctgatggcgg tttcccatct aaccgaatcc atgaaccgat
     5761 accgggaagg gaagggagac aagcccggcc gcgtgttccg tccacacgtt gcggacgtac
     5821 tcaagttctg ccggcgagcc gatggcggaa agcagaaaga cgacctggta gaaacctgca
     5881 ttcggttaaa caccacgcac gttgccatgc agcgtacgaa gaaggccaag aacggccgcc
     5941 tggtgacggt atccgagggt gaagccttga ttagccgcta caagatcgta aagagcgaaa
     6001 ccgggcggcc ggagtacatc gagatcgagc tagctgattg gatgtaccgc gagatcacag
     6061 aaggcaagaa cccggacgtg ctgacggttc accccgatta ctttttgatc gatcccggca
     6121 tcggccgttt tctctaccgc ctggcacgcc gcgccgcagg caaggcagaa gccagatggt
     6181 tgttcaagac gatctacgaa cgcagtggca gcgccggaga gttcaagaag ttctgtttca
     6241 ccgtgcgcaa gctgatcggg tcaaatgacc tgccggagta cgatttgaag gaggaggcgg
     6301 ggcaggctgg cccgatccta gtcatgcgct accgcaacct gatcgagggc gaagcatccg
     6361 ccggttccta atgtacggag cagatgctag ggcaaattgc cctagcaggg gaaaaaggtc
     6421 gaaaaggtct ctttcctgtg gatagcacgt acattgggaa cccaaagccg tacattggga
     6481 accggaaccc gtacattggg aacccaaagc cgtacattgg gaaccggtca cacatgtaag
     6541 tgactgatat aaaagagaaa aaaggcgatt tttccgccta aaactcttta aaacttatta
     6601 aaactcttaa aacccgcctg gcctgtgcat aactgtctgg ccagcgcaca gccgaagagc
     6661 tgcaaaaagc gcctaccctt cggtcgctgc gctccctacg ccccgccgct tcgcgtcggc
     6721 ctatcgcggc cgctggccgc tcaaaaatgg ctggcctacg gccaggcaat ctaccagggc
     6781 gcggacaagc cgcgccgtcg ccactcgacc gccggcgccc acatcaaggc accctgcctc
     6841 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     6901 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     6961 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc
     7021 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     7081 cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg
     7141 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     7201 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     7261 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     7321 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     7381 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     7441 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     7501 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     7561 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     7621 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     7681 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7741 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7801 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     7861 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7921 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgcattct aggtactaaa
     7981 acaattcatc cagtaaaata taatatttta ttttctccca atcaggcttg atccccagta
     8041 agtcaaaaaa tagctcgaca tactgttctt ccccgatatc ctccctgatc gaccggacgc
     8101 agaaggcaat gtcataccac ttgtccgccc tgccgcttct cccaagatca ataaagccac
     8161 ttactttgcc atctttcaca aagatgttgc tgtctcccag gtcgccgtgg gaaaagacaa
     8221 gttcctcttc gggcttttcc gtctttaaaa aatcatacag ctcgcgcgga tctttaaatg
     8281 gagtgtcttc ttcccagttt tcgcaatcca catcggccag atcgttattc agtaagtaat
     8341 ccaattcggc taagcggctg tctaagctat tcgtataggg acaatccgat atgtcgatgg
     8401 agtgaaagag cctgatgcac tccgcataca gctcgataat cttttcaggg ctttgttcat
     8461 cttcatactc ttccgagcaa aggacgccat cggcctcact catgagcaga ttgctccagc
     8521 catcatgccg ttcaaagtgc aggacctttg gaacaggcag ctttccttcc agccatagca
     8581 tcatgtcctt ttcccgttcc acatcatagg tggtcccttt ataccggctg tccgtcattt
     8641 ttaaatatag gttttcattt tctcccacca gcttatatac cttagcagga gacattcctt
     8701 ccgtatcttt tacgcagcgg tatttttcga tcagtttttt caattccggt gatattctca
     8761 ttttagccat ttattatttc cttcctcttt tctacagtat ttaaagatac cccaagaagc
     8821 taattataac aagacgaact ccaattcact gttccttgca ttctaaaacc ttaaatacca
     8881 gaaaacagct ttttcaaagt tgttttcaaa gttggcgtat aacatagtat cgacggagcc
     8941 gattttgaaa ccgcggtgat cacaggcagc aacgctctgt catcgttaca atcaacatgc
     9001 taccctccgc gagatcatcc gtgtttcaaa cccggcagct tagttgccgt tcttccgaat
     9061 agcatcggta acatgagcaa agtctgccgc cttacaacgg ctctcccgct gacgccgtcc
     9121 cggactgatg ggctgcctgt atcgagtggt gattttgtgc cgagctgccg gtcggggagc
     9181 tgttggctgg ctgg
//
