LOCUS       sequence-370643-        5348 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Gateway entry vector for the expression of mCherry and OVA323-339 
            peptide fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5348)
  TITLE     Gateway plasmids for mammalian expression of mCherry-OVA(OTIIpep)
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5348)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pENTR-mCherry.OVA(323-339)
FEATURES             Location/Qualifiers
     source          1..5348
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(75..102)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(194..280)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    333..432
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      435..444
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             441..1148
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(586..604)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     854..873
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(1222..1321)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1433..1452)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1444..2253
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1511..1530)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2396..2984
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2885..2904
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgctag ccaggaagag
       61 tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg
      121 cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat
      181 ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
      241 gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact
      301 ctcgcgttaa cgctagcatg gatctcgggc cccaaataat gattttattt tgactgatag
      361 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
      421 aaaaagcagg cttcgccacc atggtgagca agggcgagga ggataacatg gccatcatca
      481 aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga
      541 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
      601 ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct
      661 ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg
      721 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc
      781 aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact
      841 tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc
      901 ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg
      961 acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc
     1021 tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca
     1081 ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc
     1141 tgtacaagtc agcagagagc ctgaagatat ctcaagctgt ccatgcagca catgcagaaa
     1201 tcaatgaagc aggcagataa gacccagctt tcttgtacaa agttggcatt ataagaaagc
     1261 attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt
     1321 gccatccagc tgcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat
     1381 aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt
     1441 gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct
     1501 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     1561 cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     1621 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     1681 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     1741 cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     1801 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     1861 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     1921 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     1981 atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     2041 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     2101 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     2161 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     2221 cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac
     2281 tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac
     2341 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     2401 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     2461 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     2521 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     2581 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     2641 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     2701 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     2761 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     2821 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     2881 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     2941 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3001 cctttttacg gttcctggcc ttttgctggc cttttgctca ctccctgctt tgtatgaaga
     3061 acgccgttcg ctgtcgtcag gttgtggtcg acgcctcttg ctgtctctgc cttggtctgt
     3121 gcggtgtcga tgtgagctgg accccaactg tggactgtcc ccttcagctt ctctatgcct
     3181 gcttgacttg tgaccatgcc tgttctcacg tgcaactcta tcgcggtctt gcctgtgtga
     3241 atgtgacttt ctctctcgac gctcatagct atgacggtcc ctgtctcgtc tgctgctatg
     3301 tgacggactt gattcgcgaa gcggcgatga ggacctcctg tcttttcggc aagactcttc
     3361 gcccttcgag ctggttgcct gcatcctcga ctcggacatg atgatgttga tatcgagcac
     3421 gttgatggga tcgatcctag aatatcaatc gagccaccgt caatttcgag cgctaagtgg
     3481 agttgtccga gtcccgaatc acaagaatta ggctcgtgct ctgtgagatc tctcgcgaaa
     3541 cccagatgaa ggaaaaaaat cggaagatcg cggaagaagt ggggttcgca tggtctaaca
     3601 ttgtcgcatt cctcacagtt tcggctggaa acggcaggga caatcacgag aaatgcgcga
     3661 cgagtattca gttgtccaaa attatgtcag cttgaaagtt ggaaacggcg gaagaaaatt
     3721 cgtcaggagg gcgttgcggc tagggtgaag aacggagaca atttcggctt tgaaattggc
     3781 tcgactctga ttggatcatg acgcactgct ggtccacaac cgtgtcgaag gtgccgtctt
     3841 tactacaggt ccgctggaag caaaatggaa aaagccgctg gagcccgact acagagccgc
     3901 cgtgttttgg taatcagtcg gcaaataggt cagcacagcg cagcgtgaca ggttcttgca
     3961 atttacagca cagctcgtcc gtctacgact ttgcacacca caaagtgtgc ggggagcaaa
     4021 ggagccgatc ttggtcgcgc gcaaagccaa ggagtcttga acctgagagt gtgcgtgtct
     4081 tgtgacgctt gcccttctgt actttgctgt gacactacca ccacatctgt cttggctttt
     4141 tgttcataca tccacaccga ccatgtcgct attcaacgtc agcaacggtc ttcgtaccgc
     4201 tctccgacct tctgttgcca gctcttcgcg cgttgctgcc ttttccacaa ccgccgctgc
     4261 ccgtctcgcc acacccacct ctgacaacgt tggcagttcg ggcaagcctc agcacttgaa
     4321 gcagttcaag atctaccgat ggaaccctga caagccctcg gagaagcctc gtctgcagtc
     4381 gtacacactg gacctcaacc agaccggtcc aatggttctc gacgcgctca tcaaaatcaa
     4441 gaacgaaatt gaccctacgc tcaccttccg tcgctcgtgc cgtgagggta tctgcggttc
     4501 gtgcgctatg aatattgacg gtgtcaacac cctcgcctgc ctctgccgga tcgacaagca
     4561 gaatgacacc aagatctacc ccttgccgca catgtacatt gtcaaggacc tcgtgccaga
     4621 cttgacccag ttctacaagc agtaccgatc catcgagcct ttcctcaagt ccaacaacac
     4681 cccttctgag ggtgaacatc ttcagtcgcc cgaggagcgt cgtcgactcg acggtctgta
     4741 cgagtgcatt ctgtgcgcgt gctgctccac atcctgcccc tcttactggt ggaatcagga
     4801 cgagtacctt ggccccgccg tgctcatgca ggcgtaccga tggatggccg actcgcgtga
     4861 cgactttggt gaggagcgaa gacagaagct cgagaacacc ttttcgctct accgatgcct
     4921 taccatcatg aactgctcca ggacctgccc caagaacctc aaccctggta aggcaattgc
     4981 acagatcaag aaggacatgg ccgtcggcgc acccaaggct tccgagcgcc ctatcatggc
     5041 ttcgtcgtaa tcttgatata tcatatcgtt ctttcctcag cacttctttt gtcaatttca
     5101 aaagtatcta attgcattca actccgcttg tggtttgttg ttcagtgaga gtggaaacgc
     5161 tacgggcaag atgagggcag tgttctggcg acggaaaagt gtgcaagtgt ctggcctgcg
     5221 tcctcgctgg ttccagcagc cgatgcagga cgtgtaccta gcgatttctt cgacagccta
     5281 ttgtggcagc cgcgattcgc cacaatcgta cgaaagcgag acgagttgag cgaagatccc
     5341 cgggcgag
//
