LOCUS       sequence-390640-        4790 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enables recombination mediated gene delivery into Agrobacterium 
            strains for subsequent transformation of plants.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4790)
  AUTHORS   Ferguson S, Abel NB, Reid D, Madsen LH, Luu TB, Andersen KR, 
            Stougaard J, Radutoiu S
  TITLE     A simple and efficient protocol for generating transgenic hairy 
            roots using Agrobacterium rhizogenes.
  JOURNAL   PLoS One. 2023 Nov 1;18(11):e0291680. doi: 
            10.1371/journal.pone.0291680. eCollection 2023.
  PUBMED    37910566
REFERENCE   2  (bases 1 to 4790)
  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: "PLoS One.";
            date: "2023-11-1"; volume: "18(11):e0291680. doi"; pages: " 
            10.1371/journal.pone.0291680. eCollection 2023"
COMMENT     Sequence Label: pIV101
FEATURES             Location/Qualifiers
     source          1..4790
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(15..33)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        101..205
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(206..274,275..1066)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(424..443)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1237..1825
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1726..1745
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1979..1996
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2011..2151
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2237..2259
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             3014..3805
                     /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"
     oriT            3931..4040
                     /note="incP origin of transfer"
     protein_bind    4243..4264
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4279..4284,4285..4302,4303..4309)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4317..4333
                     /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)."
     primer_bind     4322..4344
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4341..4357
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4341..4357
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4353..4676
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    4370..4426
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(4427..4444)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4427..4443)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4436..4458)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ttgaagtcga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
       61 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
      121 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
      181 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
      241 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
      301 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
      361 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
      421 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
      481 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
      541 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
      601 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
      661 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
      721 aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt
      781 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
      841 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
      901 atctggagcc ggtgagcgtg gctctcgcgg tatcattgca gcactggggc cagatggtaa
      961 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     1021 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     1081 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     1141 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     1201 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     1261 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     1321 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     1381 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     1441 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     1501 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     1561 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     1621 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     1681 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     1741 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     1801 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     1861 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     1921 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     1981 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
     2041 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
     2101 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     2161 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     2221 caagctgtga ccgactccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     2281 cgcgcgaggc agctgctcgc gcaggctggg tgccaagctc tcgggtaaca tcaaggcccg
     2341 atccttggag cccttgccct cccgcacgat gatcgtgccg tgatcgaaat ccagatcctt
     2401 gacccgcagt tgcaaaccct cactgatccg catgcccgtt ccatacagaa gctgggcgaa
     2461 caaacgatgc tcgccttcca gaaaaccgag gatgcgaacc acttcatccg gggtcagcac
     2521 caccggcaag cgccgcgacg gccgaggtgt tccgatctcc tgaagccagg gcagatccgt
     2581 gcacagcacc ttgccgtaga agaacagcaa ggccgccaat gcctgacgat gcgtgcagac
     2641 cgaaaccttg cgctcgttcg ccagccagga cagaaatgcc tcgacttcgc tgctgcccaa
     2701 ggttgccggg tgacgcacac cgtggaaacg gatgaaggca cgaacccagt ggacataagc
     2761 ctgttcggtt cgtaagctgt aatgcaagta gcgtatgcgc tcacgcaact ggtccagaac
     2821 cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt gttatgactg
     2881 tttttttggg gtacagtcta tgcctcgggc atccaagcag caagcgcgtt acgccgtggg
     2941 tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagggcagtc gccctaaaac
     3001 aaagttaaac atcatgaggg aagcggtgat cgccgaagta tcgactcaac tatcagaggt
     3061 agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt tgtacggctc
     3121 cgcagtggat ggcggcctga agccacacag tgatattgat ttgctggtta cggtgaccgt
     3181 aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa cttcggcttc
     3241 ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc acgacgacat
     3301 cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc agcgcaatga
     3361 cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta tcttgctgac
     3421 aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac tctttgatcc
     3481 ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc
     3541 gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca tttggtacag
     3601 cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct
     3661 gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg gacaagaaga
     3721 agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccactacg tgaaaggcga
     3781 gatcaccaag gtagtcggca aataatgtct aacaattcgt tcaagccgac gccgcttcgc
     3841 ggcgcggctt aactcaagcg ttagatgcac taagcacata attgctcaca gccaaactat
     3901 caggtcaagt ctgcttttat tatttttaag ccggccagcc tcgcagagca ggattcccgt
     3961 tgagcaccgc caggtgcgaa taagggacag tgaagaagga acacccgctc gcgggtgggc
     4021 ctacttcacc tatcctgccc cgtgcataat aagccctaca caaattggga gatatatcat
     4081 gaaaggctgg ctttttcttg ttatcgcaat agttggcgaa gtaatcgcaa catccgcatt
     4141 aaaatctagc gagggcttta cttgccttgt cttctgggag tgagaccgca gctggcacga
     4201 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     4261 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     4321 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcttgcatg
     4381 cctgcaggtc gactctagag gatccccggg taccgagctc gaattcactg gccgtcgttt
     4441 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     4501 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     4561 tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg
     4621 gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     4681 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     4741 catccgctta cagacaagct gtgacggtct cacgctacga agacaaggga
//
