LOCUS       sequence-430579-        3066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry Vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3066)
  AUTHORS   Sivakumar P, Vaishnavi V, Gayatri K, Satheesh GR, Siddiqi I
  TITLE     Improved validation of protein interactions using bicistronic BiFC 
            (Bi2FC).
  JOURNAL   Physiol Mol Biol Plants. 2024 Jul;30(7):1047-1054. doi: 
            10.1007/s12298-024-01477-y. Epub 2024 Jul 2.
  PUBMED    39100877
REFERENCE   2  (bases 1 to 3066)
  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"; doi: 
            "10.1007/s12298-024-01477-y"; journalName: "Physiol Mol Biol 
            Plants"; date: "2024-07"; volume: "30"; issue: "7"; pages: 
            "1047-1054"
COMMENT     Sequence Label: pCENTR_MGGAGM
FEATURES             Location/Qualifiers
     source          1..3066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        182..259
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(279..298)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     terminator      complement(648..675)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(767..853)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     902..924
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     916..933
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     917..933
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    949..1048
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1070..1169)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1194..1212)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1280..1384
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1385..1453,1454..2245)
                     /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(1603..1622)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2416..3004
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2905..2924
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcataaattc
      181 cgacaccatc gaatggcgca aaacctttcg cggtatggca tgatagcgcc cggaagagag
      241 tcaattcagg gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg
      301 tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac
      361 gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca
      421 acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca
      481 cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt
      541 ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct
      601 tctcgcgcaa cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg
      661 tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc
      721 accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca
      781 ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct
      841 ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt
      901 tcccagtcac gacgttgtaa aacgacggcc agtcttaagc tcgggcccca aataatgatt
      961 ttattttgac tgatagtgac ctgttcgttg caacaaattg atgagcaatg cttttttata
     1021 atgccaactt tgtacaaaaa agcaggctta atgggtggtg caggaatgaa cccagctttc
     1081 ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc
     1141 actatcagtc aaaataaaat cattatttgc catccagctg ataagacgaa agggcctcgt
     1201 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     1261 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     1321 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     1381 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     1441 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     1501 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     1561 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     1621 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     1681 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     1741 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     1801 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     1861 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     1921 gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     1981 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     2041 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     2101 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     2161 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     2221 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     2281 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     2341 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     2401 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2461 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2521 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     2581 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2641 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2701 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2761 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2821 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2881 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2941 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3001 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3061 catgtt
//
