LOCUS       sequence-420901-        2623 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Gateway Donor Vector with attP1 and attP2 sites modified from 
            PDONR221 and Ampicillin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2623)
  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 2623)
  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: pCONR221
FEATURES             Location/Qualifiers
     source          1..2623
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     53..75
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     67..84
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     68..84
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    101..332
                     /gene="mutant version of attP"
                     /label=attP1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(728..1033)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(830..849)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1353..2036)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     primer_bind     1951..1970
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2037..2139)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(2284..2515)
                     /gene="mutant version of attP"
                     /label=attP2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2541..2559)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tttgatgcct ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca
       61 cgacgttgta aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga
      121 ctgatagtga cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact
      181 ttgtacaaaa aagctgaacg agaaacgtaa aatgatataa atatcaatat attaaattag
      241 attttgcata aaaaacagac tacataatac tgtaaaacac aacatatcca gtcactatga
      301 atcaactact tagatggtat tagtgacctg tagtcgaccg acagccttcc aaatgttctt
      361 cgggtgatgc tgccaactta gtcgaccgac agccttccaa atgttcttct caaacggaat
      421 cgtcgtatcc agcctactcg ctattgtcct caatgccgta ttaaatcata aaaagaaata
      481 agaaaaagag gtgcgagcct cttttttgtg tgacaaaata aaaacatcta cctattcata
      541 tacgctagtg tcatagtcct gaaaatcatc tgcatcaaga acaatttcac aactcttata
      601 cttttctctt acaagtcgtt cggcttcatc tggattttca gcctctatac ttactaaacg
      661 tgataaagtt tctgtaattt ctactgtatc gacctgcaga ctggctgtgt ataagggagc
      721 ctgacattta tattccccag aacatcaggt taatggcgtt tttgatgtca ttttcgcggt
      781 ggctgagatc agccacttct tccccgataa cggagaccgg cacactggcc atatcggtgg
      841 tcatcatgcg ccagctttca tccccgatat gcaccaccgg gtaaagttca cgggagactt
      901 tatctgacag cagacgtgca ctggccaggg ggatcaccat ccgtcgcccg ggcgtgtcaa
      961 taatatcact ctgtacatcc acaaacagac gataacggct ctctctttta taggtgtaaa
     1021 ccttaaactg catttcacca gcccctgttc tcgtcagcaa aagagccgtt catttcaata
     1081 aaccgggcga cctcagccat cccttcctga ttttccgctt tccagcgttc ggcacgcaga
     1141 cgacgggctt cattctgcat ggttgtgctt accagaccgg agatattgac atcatatatg
     1201 ccttgagcaa ctgatagctg tcgctgtcaa ctgtcactgt aatacgctgc ttcatagcat
     1261 acctcttttt gacatacttc gggtatacat atcagtatat attcttatac cgcaaaaatc
     1321 agcgcgcaaa tacgcatact gttatctggc ttttagtaag ccggatccac gcggcgttta
     1381 cgccccccct gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg
     1441 aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct
     1501 tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg
     1561 tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca
     1621 ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat
     1681 atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca
     1741 gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg
     1801 tctttcattg ccatacggaa ttccggatga gcattcatca ggcgggcaag aatgtgaata
     1861 aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc
     1921 agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct
     1981 ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta
     2041 gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt
     2101 tcattatggt gaaagttgga acctcttacg tgccgatcaa cgtctcattt tcgccaaaag
     2161 ttggcccagg gcttcccggt atcaacaggg acaccaggat ttatttattc tgcgaagtga
     2221 tcttccgtca caggtattta ttcggcgcaa agtgcgtcgg gtgatgctgc caacttagtc
     2281 gactacaggt cactaatacc atctaagtag ttgattcata gtgactggat atgttgtgtt
     2341 ttacagtatt atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata
     2401 tcattttacg tttctcgttc agctttcttg tacaaagttg gcattataag aaagcattgc
     2461 ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat
     2521 ccagctgata agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     2581 aataatggtt tcttagacgt caggtggcac ttttcgggga aat
//
